Volume / tomas 28(3) - SodininkystÄ ir darÅ¾ininkystÄ - SodininkystÄs ...

Lietuvos sodininkystĖs ir darŽininkystĖs instituto ir 

lietuvos ŽemĖs Ūkio universiteto mokslo darbai 

Scientific works of the Lithuanian institute of 

horticulture AND lITHUANIAN UNIVERSITY OF AGRICULTURE 

SodininkystĖ ir darŽininkystĖ 

28(3) 

Eina nuo 1983 m. 

Published since 1983 

Babtai 2009

UDK 634/635 (06) 

Redaktorių kolegija 

Editorial Board 

Doc. dr. Česlovas BOBINAS – pirmininkas (LSDI, biomedicinos mokslai, agronomija), 

prof. habil. dr. Pavelas DUCHOVSKIS (LSDI, biomedicinos mokslai, agronomija), 

dr. Edite KAUFMANE (Latvija, Dobelės sodo augalų selekcijos stotis, 

biomedicinos mokslai, biologija), 

dr. Aleksandras KMITAS (LŽŪU, biomedicinos mokslai, agronomija), 

dr. Laimutis RAUDONIS (LSDI, biomedicinos mokslai, agronomija), 

prof. habil. dr. Vidmantas STANYS (LSDI, biomedicinos mokslai, agronomija), 

prof. habil. dr. Andrzej SADOWSKI (Varšuvos ŽŪA, biomedicinos mokslai, agronomija), 

dr. Audrius SASNAUSKAS (LSDI, biomedicinos mokslai, agronomija), 

prof. habil. dr. Algirdas SLIESARAVIČIUS (LŽŪU, biomedicinos mokslai, agronomija). 

Redakcinė mokslinė taryba 

Editorial Scientific Council 

Doc. dr. Česlovas BOBINAS – pirmininkas (Lietuva), 

prof. habil. dr. Pavelas DUCHOVSKIS (Lietuva), dr. Kalju KASK (Estija), 

dr. Edite KAUFMANE (Latvija), prof. habil. dr. Zdzisław KAWECKI (Lenkija), 

prof. habil.dr. Albinas LUGAUSKAS (Lietuva), habil. dr. Maria LEJA (Lenkija), 

prof. habil. dr. Lech MICHALCZUK (Lenkija), prof. habil. dr. Andrzej SADOWSKI (Lenkija), 

dr. Audrius SASNAUSKAS (Lietuva), prof. dr. Ala SILAJEVA (Ukraina), 

prof. habil. dr. Algirdas SLIESARAVIČIUS (Lietuva), 

prof. habil. dr. Vidmantas STANYS (Lietuva), 

prof. dr. Viktor TRAJKOVSKI (Švedija). 

Redakcijos adresas: 

Lietuvos sodininkystės ir daržininkystės institutas 

LT-54333 Babtai, Kauno r. 

Tel. (8 37) 55 52 10 

Faksas (8 37) 55 51 76 

El. paštas institutas@lsdi.lt 

Address of the Editorial Office: 

Lithuanian Institute of Horticulture 

LT-54333 Babtai, Kaunas district, Lithuania 

Phone: +370 37 55 52 10 

Telefax: +370 37 55 51 76 

E-mail institutas@lsdi.lt 

Leidinio adresas internete www.lsdi.lt 

Leidinys cituojamas CAB Abstracts, EBSCO Publishing, VINITI duomenų bazėse 

ISSN 0236-4212 © Lietuvos sodininkystės ir daržininkystės institutas, 2009 

© Lietuvos žemės ūkio universitetas, 2009

SCIENTIFIC WORKS OF THE LITHUANIAN INSTITUTE OF 

HORTICULTURE AND LITHUANIAN UNIVERSITY OF AGRICULTURE. 

SODININKYSTĖ IR DARŽININKYSTĖ. 2009. 28(3). 

Controling pear psylla with Abamectin in Bulgaria 

Veselin Arnaudov, Hristina Kutinkova 

Institute of Fruit Growing, “Ostromila” 12, 4004 Plovdiv, Bulgaria, 

e-mail vaarnaudov@abv.bg 

Considering pear psylla (Cacopsylla pyri L.) resistance to insecticides routinely used in 

Bulgaria the study was undertaken aimed at improving the system of this pest control. The 

experiments were carried out in Plovdiv region, South-Central Bulgaria on ‘Buttira Precoce 

Morettini’ and ‘Beurre Hardy’ pear trees in 2007–2008. Efficacy of a. i. Abamectin of a pesticide 

supposed to be more selective, not harmful to beneficial fauna, was tested against the 

background of a. i. Amitraz as commonly used insecto-acaricide. Post-bloom applications 

of Abamectin provide a significant control of summer populations of pear psylla. There are 

needed two consecutive sprays of Abamectin at the rate of 240 g a. i. per ha applied after 

bloom on young nymphs of the second generation. These treatments do not kill summer adult 

forms; however, cause a significant reduction in density of summer pear psylla eggs and 

nymphs. Abamectin may be recommended for the integrated pest management programmes 

in pear production. 

Key words: Abamectin, adults, Amitraz, beneficial fauna, Cacopsylla pyri, eggs, efficacy, 

nymphs, summer populations. 

Introduction. Two psyllid species Cacopsylla pyri (L.) and Cacopsylla pyrisuga 

(Foster) (Hemiptera: Psyllidae) have been reported to cause damage to pear trees in 

fruit growing regions of Bulgaria; however, only C. pyri is considered as the species 

of economic importance in commercial pear orchards (Harizanov, 1966). C. pyri is 

key pest of pear in Bulgaria as well as in other European countries. Its extent and 

intensity of damage caused in Bulgarian orchards over the last 20 years has considerably 

increased. The pear psylla was particularly abundant in localities, which in 

the past had been heavily sprayed with non-selective insecticides – probably due to 

devastation of the natural enemies. 

Pear psylla is difficult to control because of its ability to develop resistance to 

insecticides from various groups, including organophosphates and pyrethroids. Since 

the mid-1980’s, populations of C. pyri in Bulgaria, as well as in many other European 

countries were found to be resistant to a broad spectrum of insecticides (Harizanov, 

1982; Berrada et al., 1995; Arnauodov, 2001; Buès, Boudinhon, 2002; Buès et al., 

2003). Cross-resistance to organophosphates, pyrethroids and Amitraz has been found 

in populations of Cacopsylla pyri in Europe (Buès et al., 1999). Kocourek and Stará 

3

(2006) reported on resistance of C. pyri populations originated from an orchard intensely 

treated with Nomolt 15 SC (teflubenzuron) in Czech Republic. 

According to many growers in Bulgaria, treatments with different insecticides 

do not provide an expected control of C. pyri, with the exception of insecto-acaricide 

Amitraz. Amitraz was the only product that lately ensured an adequate protection 

from C. pyri in our country. After prohibiting its use in 2005, the most common active 

ingredient involved is Abamectin, a mixture of 80 % avermectin B-1 a and 20 % 

avermectin B-1 b, as alternative means for control of the pest. 

Biological control by beneficial entomo- and acarofauna present in the orchards 

offers at least a partial solution of the problem; however, the use of non-selective 

pesticides greatly reduces effectiveness of predators and parasitoids (Burts, Beers, 

1994). Their substitution by soft or selective pesticides allows more effective biological 

control (Burts, Evereti, 1983). Abamectin is high effective against pear psylla 

and soft to some beneficial arthropods, naturally controlling population of this pest 

(Nguyen, Berrada, 1994). 

The aim of the present study was to assess the efficacy of Abamectin used for 

control of Cacopsylla pyri, in order to improve the system of this pest control. 

Object, methods and conditions. Field studies were carried out in the region of 

Plovdiv in a commercial orchard, on ‘Buttira Precoce Morettini’ and ‘Beurre Hardy’ 

pear trees in 2007–2008. Efficacy of Abamectin in controlling Cacopsylla pyri was 

compared with that of Amitraz, traditionally used insecto-acaricide. Both insecticides 

were applied at the rates recommended by their manufacturers: Lirosect® 2 EC 

(20 g/l Abamectin) at a dose 1.2 l per ha (240 g a. i. per ha) and Mitac® 20 EC (20 % 

amritaz) at a dose 2.0 l per ha (400 g a. i. per ha). The insecticides were applied twice, 

at post-bloom against eggs and newly hatched nymphs of the second generation of 

C. pyri – on May 3 and May 13 in 2007 and on May 9 and May 19 in 2008. The insecticides 

were applied with “Perla 9” sprayer, using 1 000 L of liquid per ha. 

The experiments were set up a block random scheme in 3 variants and 3 replicates 

(by four trees for each replicate) for each variant, on 6-year-old pear trees with 

spindle-shaped form, from ‘Beurre Hardy’ cultivar. 

The results are given through a direct counting of living nymphs and vital eggs 

on 20 fruit clusters per replicate, including adjacent leaf rosettes and shoots (5 fruit 

clusters per tree) sampled from different parts of tree crowns. The evaluation was 

carried out before the first treatment and on the 3 rd and 9 th day after each insecticide 

treatment by inspecting the sampled fruit clusters in the laboratory by binocular and 

registering the average number of eggs or nymphs per cluster. 

Biological efficacy was expressed as a percentage, and was calculated according 

to the formula of Henderson and Tilton (1955). The data were statistically analyzed 

by Duncan’s test (Steele, Torrie, 1980). 

Results. In 2007, Abamectin (Lirosect ® 2 EC) applied twice in the post-bloom 

period against the second generation of C. pyri demonstrated a high biological efficacy 

against newly hatched nymphs of this pest (on the average 87.6–79.9 %), what 

was close to the efficacy of Amitraz (Mitac ® 20 EC) varying from 87.1 to 66.4 % on 

the average (Table 1). 

4

Table 1. Efficacy of the pesticides tested for control of Cacopsylla pyri nymphs 

in 2007 

1 lentelė. Cacopsylla pyri nimfų naikinimui naudotų pesticidų efektyvumas, 2007 m. 

Insecticides 

Insekticidai 

02.05 

T - 1 

Number of nymphs of C. pyri on average per fruit cluster and 

biological efficacy of the insecticides 

Vidutinis C. pyri nimfų skaičius ant vaisių kekės ir 

biologinis insekticidų efektyvumas (%) 

06.05 

T + 3 

12.05 

T + 9 

5 

13.05 

T 

16.05 

T + 3 

22.05 

T + 9 

efficacy 

efektyvumas 

(%) 06–22.05 

Number of nymphs of C. pyri 

C. pyri nimfų skaičius 

Lirosect ® 2 EC 21.8 1.5 26.2 - 1.5 16.6 - 

Mitac ® 20 EC 17.5 5.4 14.7 - 6.8 10.8 - 

Control 20.5 62.7 109.3 - 210.3 186.4 - 

Kontrolė 

Biological efficacy 

Biologinis efektyvumas (%) 

Lirosect ® 2 EC - 97.75 a 77.46 a - 97.02 a 62.85 a 84.52 a 

Mitac ® 20 EC - 89.91 a 84.25 a - 75.96 b 56.92 b 76.76 a 

The means followed by the same letter do not differ significantly from one another (p = 0.05) 

Ta pačia raide pažymėtos reikšmės patikimai viena nuo kitos nesiskiria (p = 0,05) 

Similar results were obtained in 2008. Аbamectin applied twice after bloom 

showed a high biological efficacy against newly born nymphs of C. pyri – on the average 

82.8–75.6 % (Table 2). It was nearly identical to the efficacy of Amitraz (on the average 

81.0–54.4 %). Тhe efficacy of tested insecticides against newly hatched nymphs of 

C. pyri varied throughout the entire period of study on the average between 84.5 and 

76.8 % for Abamectin and between on the average 79.2 and 67.7 % for Amitraz. 

Table 2. Efficacy of the pesticides tested for control of Cacopsylla pyri nymphs 

in 2008 

2 lentelė. Cacopsylla pyri nimfų naikinimui naudotų pesticidų efektyvumas, 2008 m. 

Insecticides 

Insekticidai 

Number of nymphs of C. pyri in average per fruit cluster and biological 

efficacy of the insecticides 

Vidutinis C. pyri nimfų skaičius ant vaisių kekės ir biologinis insekticidų efektyvumas 

(%) 

08.05 

T - 1 

11.05 

T + 3 

18.05 

T + 9 

19.05 

T 

22.05 

T + 3 

28.05 

T + 9 

efficacy 

efektyvumas 

(%) 11–28.05 

1 2 3 4 5 6 7 8 

Number of nymphs of C. pyri 

C. pyri nimfų skaičius 

Lirosect ® 2 EC 9.5 0 49.8 - 0.8 13.8 - 

Mitac ® 20 EC 7.5 1.0 28.0 - 3.6 19.0 - 

Control 

Kontrolė 

8.0 33.0 86.1 - 110.7 72 -

Table 2 continued 

2 lentelės tęsinys 

1 2 3 4 5 6 7 8 



Lirosect ® 2 EC - 100 a 51.25 a - 98.75 a 66.86 a 79.21 a 

Mitac ® 20 EC - 96.77 a 65.31 a - 90.00 a 18.85 b 67.73 a 



Abamectin was very different in respect to its insecticidal properties. Comparing 

this compound with Amitraz, a faster initial effect of Abamectin was noted. Its persistence 

was relatively shorter. Abamectin apparently remained active in leaf tissues 

for not more than 7–10 days. It doesn’t kill summer adults, albeit caused significant 

reductions in the density of summer pear psylla eggs and nymphs. The first 2-instar 

nymphs of C. pyri were much more susceptible to both insecticides than the 3–5-instar 

nymphs of the pest. 

The biological efficacy of tested insecticides against the C. pyri eggs ranged in 

general for Abamectin from 62.6 to 13.9 % in 2007 and from 63.5 to 19.2 % in 2008, 

whereas for Amitraz from 58.3 to 7.0 % in 2007 and from 39.0 to 9.1 % in 2008. 

Compared to Amitraz, Abamectin demonstrated a higher biological efficacy against 

the C. pyri eggs (Tables 3 and 4). 

Table 3. Efficacy of the pesticides tested for control of Cacopsylla pyri eggs in 

2007 

3 lentelė. Cacopsylla pyri kiaušinėlių naikinimui naudotų pesticidų efektyvumas, 

2007 m. 

Insecticides 

Insekticidai 

Number of eggs of C. pyri in average per fruit cluster and biological 

efficacy of the insecticides 

Vidutinis C. pyri kiaušinėlių skaičius ant vaisių kekės ir 

biologinis insekticidų efektyvumas (%) 

02.05 

T-1 

06.05 

T+3 

12.05 

T+9 

13.05 

T 

16.05 

T + 3 

22.05 

T + 9 

efficacy 

efektyvumas 

(%) 06–22.05 

Number of eggs of C. pyri 

C. pyri kiaušinėlių skaičius 

Lirosect ® 2 EC 141 216 129 - 90 45 - 

Mitac ® 20 EC 156 260 166 - 138 87 - 

Control 113 357 393 - 380 90 - 

Kontrolė 



Lirosect ® 2 EC 51.51 a 73.69 a - 27.85 a 0 a 38.26 a 

Mitac ® 20 EC 47.25 b 69.40 b - 14.02 b 0 a 32.92 b 



6

Table 4. Efficacy of the pesticides tested for control of Cacopsylla pyri eggs in 

2008 

4 lentelė. Cacopsylla pyri kiaušinėlių naikinimui naudotų pesticidų efektyvumas, 

2008 m. 

Insecticides 

Insekticidai 

Number of eggs of C. pyri in average per fruit cluster and biological efficacy 

of the insecticides 

Vidutinis C. pyri kiaušinėlių skaičius ant vaisių kekės ir biologinis insekticidų 

efektyvumas (%) 

08.05 

T - 1 

11.05 

T + 3 

18.05 

T + 9 

19.05 

T 

22.05 

T + 3 

28.05 

T + 9 

efficacy 

efektyvumas 

(%) 11–28.05 

Number of eggs of C. pyri 

C. pyri kiaušinėlių skaičius 

Lirosect ® 2 EC 92 150 69 - 41 19 - 

Mitac ® 20 EC 68 181 90 - 71 37 - 

Control 49 155 171 - 165 39 - 

Kontrolė 



Lirosect ® 2 EC - 48.46 a 78.51 a - 38.42 a 0 a 41.35 a 

Mitac ® 20 EC - 15.85 b 62.07 b - 18.24 b 0 a 24.04 b 



Тhe efficacy of tested insecticides against the C. pyri eggs varied throughout the 

period of study on the average between 41.4 and 38.3 % for Abamectin and between 

32.9 and 24 % for Amitraz. 

Discussion. The control of the second summer generation of C. pyri eggs and 

larvae is very important in the seasonal programme for control of pear psylla in pear 

growing areas of Bulgaria. A field study conducted in a commercial pear orchard 

showed that the tested product Lirosect ® 2 EC (Abamectin) gave promising results 

in control of pear psylla, Cacopsylla pyri. This insecticide reduced the populations 

of C. pyri to an extent comparable with that the standard insecticide Mitac ® 20 EC 

(Amitraz), at the rates used. Two consecutive applications of Abamectin after bloom, 

on young nymphs of the second generation provide a significant control of summer 

populations of pear psylla. 

Abamectin demonstrated a high biological efficacy against pear psylla eggs and 

newly hatched nymphs, similar to that of Amitraz. Its characteristic features are quick 

initial toxicity and a relatively short persistence (not more than 7–10 days). The product 

exhibited a strong deterring effect on oviposition by summer females and hence 

caused a reduction of the total number of laid eggs. This compound applied twice 

against C. pyri eggs gave better results than the standard compound Amitraz. As far as 

Abamectin is concerned, the results obtained in this study are in agreement with those 

reported earlier by the other authors, under different conditions (Buès, Boudinhon, 

2002; Marcic, et al., 2002; Kocourek, Stará, 2006). 

7

At the concentration of 0.12 % (Lirosect ® 2 EC), the highest biological activity of 

the product was recorded against the young (1st and 2nd) nymphal stages (up to 95 % 

mortality) in comparison with the other biological stages of the pest. The insecticide 

was less active against the older 3–5-instar nymphs, causing only 35 % mortality of 

them, at the same concentration. 

So, Abamectin may be used in control of pear psylla, instead of Amitraz. 

Considering its selectivity, it is less harmful to beneficial arthropods. Therefore, it 

should be especially useful as a component of the integrated pest management (IPM) 

in pear orchards. 

Conclusions. 1. The insecto-acaricide Abamectin shows a high biological activity 

to C. psylla eggs and nymphs and may be successfully used in the seasonal control 

programme against pear psylla. 

2. Abamectin does not kill summer adult forms of pear psylla, albeit causes significant 

reduction in summer densities of pear psylla eggs and nymphs. 

3. Abamectin demonstrates a quick initial effect and relatively short persistence – 

retained only for 7–10 days. 

4. Two treatments with Abamectin at a dose of 240 g a. i. per ha, applied after 

bloom – against newly hatched nymphs of the second generation, should be applied 

for control of pear psylla. 

Gauta 2009 06 30 

Parengta spausdinti 2009 08 11 

References 

1. Arnaoudov V. 2001. Efficacy of some pesticides for control of pear psylla, Psylla 

pyri L. in Bulgaria. 9 th International Conference of Horticulture, Lednice 2001, 

1: 15–19. 

2. Berrada S., Nguyen T. X., Merzoug D., Fournier D. 1995. Selection for 

Monocrotophos resistance in pear psylla, Cacopsylla pyri (L.) (Hom, Psyllidae). 

Journal of Applied Entomology, 119(7): 507–510. 

3. Buès R., Boudinhon L. 2002. Insecticide resistance in the pear psylla. Acta 

Horticulturae (ISHS), 596: 567–570. 

4. Buès R. , Boudinhon L. , Toubon J. F. , Faivre D’Arcier F. 1999. Geographic and 

seasonal variability of resistance to insecticides in Cacopsylla pyri, L. (Hom, 

Psyllidae). Journal of Applied Entomology, 123(5): 289–297. 

5. Buès, R. , Boudinhon , L., Toubon J. F. 2003. Resistance of pear psylla Cacopsylla 

pyri L. (Hom; Psyllidae) to deltametrin and synergism with piperonil butoxide. 

Journal of Applied Entomology, 127(5): 305–312. 

6. Buès, R., Toubon J. F. , Boudinhon L. 2000. Genetic analysis of resistance to 

azinphosmethyl in the pear psylla Cacopsylla pyri. Entomologia Experimentalis 

et Applicata, 96(2): 159–166. 

8

7. Burts E. C., Evereti C. 1983. Effectiveness of a soft pesticide program on pear 

pests. Journal of Economic Entomology, 76(4): 936–941. 

8. Burts E. C., Beers E. H. 1994. Controlling pear psylla with fenoxycarb in western 

North America. OILB/SROP Bulletin, 17(2): 39–42. 

9. Harizanov A. 1966. Chemical experiments for control of the psyllids in the fruit 

crops. Horticultural and Viticultural Science, 3(2): 143–149. (in Bulgarian). 

10. Harizanov A. 1982. The pear psyllas. Plant Protection Newsletter, 4: 23–27. (in 

Bulgarian). 

11. Henderson C. F., Tilton E. W. 1955. Tests with acaricides against the brown 

wheat mite. Journal Economic Entomology, 48(2): 157–161. 

12. Kocourek F., Stará J. 2006. Journal of Fruit and Ornamental Plant Research, 14 

(Suppl. 3): 167–174. 

13. Marcic D., Peric P., Prijovic M., Ogurlic I., Andric G. 2002. Chemical control of 

Cacopsylla pyri L. in Serbian pear orchards using biorational insecticides. Acta 

Horticulturae (ISHS), 800: 941–946. 

14. Nguen T. X., Berrada, S. 1994. Toxicité relative de certains acaricides sur 

Anthocoris nemoralis, prédateur de psylles. OILB/SROP Bullletin, 17(2): 

53–56. 

15. Steel, R., Torrie, J. H. 1980. Principles and Procedures of Statistics. McGraw- 

Hill, Inc., New York. 

SODININKYSTĖ IR DARŽININKYSTĖ. MOKSLO DARBAI. 2009. 28(3). 

Kriaušių blakučių naikinimas Abamektinu Bulgarijoje 

V. Arnaudov, H. Kutinkova 

Santrauka 

Atsižvelgiant į kriaušinių blakučių (Cacopsylla pyri L.) atsparumą įprastiniams 

Bulgarijoje naudojamiems insekticidams, atliktas tyrimas, siekiant pagerinti šio kenkėjo 

naikinimo sistemą. Bandymai atlikti 2007–2008 metais Plodinovo regione, pietų-vidurio 

Bulgarijoje su ‘Buttira Precoce Morettini’ir ‘Beurre Hardy’ veislių kriaušėmis. Abamektino 

veiklioji medžiaga, kuri laikoma viena iš efektyviausių, nekenksminga naudingajai faunai, 

kurios efektyvumas palygintas su įprastai naudojama insekto-akaricido Amitrazės veikliąja 

medžiaga. Abamektino panaudojimas po žydėjimo gerokai sumažino vasarines kriaušinių 

blakučių populiacijas. Po žydėjimo ant jaunų antros kartos nimfų Abamektiną reikia purkšti 

du kartus po 240 g/ha. Šie purškimai nesunaikina vasarinių suaugusių kenkėjų, tačiau gerokai 

sumažina kriaušinių blakučių vasarinių kiaušinėlių ir nimfų gausumą. Abamektiną 

galima rekomenduoti naudoti integruotose kriaušių kenkėjų kontrolės programose. 

Reikšminiai žodžiai: Abamektinas, Amitrazas, Cacopsylla pyri, efektyvumas, kiaušinėliai, 

naudinga fauna, nimfos, suaugėliai, vasarinės populiacijos. 

9




Possibilities of integrated management of onion 

downy mildew 

Gunita Bimsteine 1 , Līga Lepse 2 , Biruta Bankina 1 

1 

Institute of Soil and Plant Sciences, Latvia University of Agriculture, 

Liela Street 2, Jelgava, LV-3004, e-mail Biruta.Bankina@llu.lv 

2 

Pūre Horticultural Research Center, Abavas Street 2, Pūre, Tukums district, 

LV-3124 Latvia, e-mail liga.lepse@puresdis.lv 

Onion downy mildew caused by Peronospora destructor is one of the most important 

onion diseases that require fungicide application. The aim of the research was to find out 

methods of integrated control of downy mildew in onion. 

Investigation was carried out in Pūre Horticultural Research Center in 2008. Three 

onion hybrids were included in the investigation: ‘Safrane’ F 1 

, ‘Hypark’ F 1 

and ‘Alonso’ F 1 

. 

There were investigated three plant protection variants: 1) fungicide applied according to the 

DACOM Plant Plus decision support system; 2) fungicide used according to spraying scheme; 

3) no fungicide was used. Fungicide with active ingredients Metalaxyl and Mankoceb were 

used in the trials. 

The first symptoms of the disease were observed only on 30 July. Differences in development 

of the disease between hybrids were detected. Severity of downy mildew achieved 1.6 

% (‘Alonso’ F 1 

), 3.1 % (‘Hypark’ F 1 

) and 4.5 % (‘Safrane’ F 1 

). 

Technical effectiveness of fungicide application fluctuated depending on varieties and 

spraying variant: 18.8–93.5 % for DACOM Plant Plus and 62.5–83.9 % for schematic spraying. 

The most effective disease control was achieved by application of DACOM programme. 

Further investigations are necessary to obtain consistent results. 

Key words: forecast, fungicides, Peronospora destructor, spraying. 

Introduction. Onion downy mildew caused by Peronospora destructor (Berk.) 

Casp. is an economically important disease causing losses both in the yield and quality 

of onion (Allium cepa L.). Infection in onion causes early defoliation, reduced size 

and poor storage ability of bulbs (Peter et al., 2004, Surviliene et al., 2008). 

The disease symptoms vary with the type of infection. Systemic infection occurs 

when plants are grown from infected bulbs, but local infection partly is caused 

by air-borne conidia. Onion bulbs with systemic infection are damaged faster and 

during moist weather all the leaf surface on infected plants is covered by grayish 

violet sporulation of P. destructor. In case of local infection, oval to cylindrical spots 

11

(3–30 mm in size) slightly paler than the rest of foliage are apparent on the leaves. 

Older leaves are attacked first and infection spreads to other leaves and plants (Peter 

et al., 2004; Palti, 1989). 

Without fungicide treatment economically significant onion production would not 

be possible. Onion downy mildew is a disease, which is effectively controlled with 

fungicide application (Whiteman, Beresford, 1998). Planting time, resistant cultivars, 

soil drainage and healthy seed or bulb material are also included in the management 

of this disease. The fungicide efficiency depends on the time of application and developmental 

stage of the disease. It is very important to notice the first symptoms of the 

disease in sufficient plant protection system (Buloviene, Surviliene, 2006; Whiteman, 

Beresford, 1998). 

Development of mildew epidemics depends primarily on moisture, but temperature 

and light also are important factors, which influence different stages of P. destructor 

(Palti, 1989, Buloviene, Surviliene, 2006). Cool temperatures (10–12 °C), moderate 

relative humidity and low solar irradiance are more favorable conditions for spore 

survival. Sporangia of P. destructor disperse mainly during the morning and early 

afternoon and they can survive in the field for several hours until the conditions are 

more suitable for infection, such as those at night (Palti, 1989; Bashi, Aylor, 1983). 

Knowledge of the relationships between weather factors and pathogen sporulation is 

important for developing predictive models of downy mildew epidemics (Palti, 1989; 

Buloviene, Surviliene, 2006). 

Different forecast models (ZWIREPO, DOWNCAST, ONIMIL and others) were 

made to improve schemes of fungicide application (Friedrich et al., 2003). Dutch farmers 

used weather based Decision support systems against diseases of field vegetables 

since the middle of the 1980 (Bouma, 2004). The Agri Yield Management system 

developed by DACOM provides growers around the world with practical solutions 

for profitable and sustainable agriculture. By combining sensor technology, internet 

and scientific knowledge, growers can continuously monitor and fine-tune their production 

process throughout the growing season (http://www.dacom.nl). It was found 

to be usable to introduce DACOM Plant Plus decision support system in the onion 

growing system in Latvia. Evaluation of the system efficiency under Latvia conditions 

in integrated plant protection system becomes interesting. 

Therefore in 2008 investigation was carried out attaining to find out the most 

effective methods of integrated control of downy mildew in onion. 

Object, methods and conditions. Investigations were carried out in Pūre Horticultural 

Research Center in 2008. Three onion hybrids were included in the investigation: 

‘Safrane’ F 1 


and ‘Alonso’ F 1 

. There were investigated three plant 

protection variants: 1) fungicide treatment was performed according to the DACOM 

Plant Plus decision support system; 2) fungicide used according to experts’ estimation 

based on experience; 3) no fungicide was used. 

Investigation was arranged in 4 replications, each plot – 10 m 2 . Onion seeds were 

sown by a precise sowing-machine “Robin Stanhay” in loamy soil on 23 April, in 

three-row beds on plane surface. Cultivation that followed was performed according 

to agro-ecological requirements of onions. 

12

Peronospora destructor control was performed with the following treatments: 

Variant 1 – Ridomil Gold MZ 68 WG (metalaxyl) 2.5 kg ha -1 on 08.07 and 14.07, 

and Penncozeb 75 DG (mankoceb) 2 kg ha -1 on 29.07. 

Variant 2 – Ridomil Gold MZ 68 WG (metalaxyl) 2.5 kg ha -1 on 14.07 and 

Penncozeb 75 DG (mankoceb) 2 kg ha -1 on 29.07. 

Onion hybrids used in the trial were characterized as follows: 

‘Safrane’ F 1 

– vegetation period in the investigation was observed shorter than it 

was given in the hybrid description – 135 days. Leaves mid-waxed, grey-green and 

slightly hanging down, average height of foliage 80 cm; 

‘Hypark’ F 1 

– vegetation period 150 days. Leaves mid-strong waxed, grey-green, 

almost upstanding, average height of foliage 78 cm; 

‘Alonso’ F 1 

– vegetation period 130 days. Leaves mid-strong waxed, green, almost 

upstanding, average height of foliage 73 cm. Resistance to Peronospora destructor is 

indicated in the description of hybrid. 

The severity of downy mildew was evaluated each week and was scored using 

5 point system: 0 – no disease symptoms; 1 – some spots of disease; 2 – damaged 

1/3 of a plant; 3 – damaged 1/2 of a plant; 4 – damaged 2/3 of a plant; 5 – all leaves 

damaged. 

Onion bulbs were harvested on the 3 rd decade of August. Yield was weighted in the 

field directly after harvest, dried in a plastic tunnel, cleaned and placed for storage. 

Meteorological data were recorded by “Lufft” automatic meteorological station. 

Air temperature, precipitation, air humidity, solar irradiation, wind and dry leave 

measurements were recorded. 

ANOVA procedures were used for experimental data processing. 

Results. First symptoms of downy mildew on the leaves of plants were noticed 

on 15 July in hybrid ‘Safrane’, but sharp development of this disease was observed 

after 30 July. Differences of downy mildew development were observed depending 

on hybrids (Fig. 1). The most susceptible hybrid was ‘Safrane’ – the severity of disease 

achieved 4.5 points. 

13

Fig. 1. Development of disease depending on fungicide application scheme 

1 pav. Ligos eiga priklausomai nuo fungicidų naudojimo schemos 

The first fungicide treatment was performed on 8 July according to DACOM 

Plant Plus decision support system, but on experience-based spraying – a week later. 

At this time disease symptoms were not observed. In total, during vegetation period 

DACOM Plant Pus recommended three applications of fungicides (Fig. 1). Treatments 

of fungicides significantly reduced the severity of downy mildew in planting, but 

efficiency was depending on hybrids as well. 

Hybrid and fungicide application scheme influenced yield. Onion yield was 

significantly higher, if scheme of DACOM Plant Plus was used (Fig. 2). Routine or 

experience-based variant did not give sufficient results. 

14

Fig. 2. Onion yield depending on hybrid and schemes of fungicide application 

2 pav. Svogūnų derliaus priklausomybė nuo hibridų ir fungicidų naudojimo schemos 

Discussion. Development of downy mildew has strong correlation with weather 

conditions. The first decade of June in 2008 was extremely dry (amount of precipitation 

only 0.1 mm), relative humidity (RH) exceeded 90 % only in some cases, but 

mostly fluctuated from 50–80 %. The most important factor for development of onion 

downy mildew is high RH, the greatest number of sporangia was produced at 100 % 

(Gilles et al., 2004), but the peak dispersal of spores coincided with drying of the leaves. 

Infection is not possible if leaves are dry (Hildebrand, Sutton, 1982). Abundant 

rainfall (60 mm) was observed in the second decade of June, but RH > 95 % only 

in July. These observations explained sharp development of the disease at the end of 

July. 

Our investigations confirm characterization of varieties according to which 

‘Alonso’ is more resistant comparing to ‘Safrane’. 

It is difficult to explain the obtained data about disease development and obtained 

yield. Significant difference between hybrids ‘Alonso’ and ‘Hypark’ in downy mildew 

severity was not observed at the end of vegetation period (12.08) depending on scheme 

of fungicide application. Visible effect of three applications of fungicide (recommended 

by DACOM Plant Plus) was noted only for hybrid ‘Safrane’. These results were 

different comparing with the other investigations. Buloviene and Surviliene observed 

high biological efficacy (74.38 % to 89.36 %) of all the used fungicides (Buloviene, 

Surviliene, 2006). 

Nevertheless, in all cases significant increase in yield was determined in trials 

with three applications of fungicide. Importance of the necessity to control the first 

symptoms of the disease (could be latent), as described by other authors (Buloviene, 

Surviliene, 2006; Battilani et al., 1996), is one of possible reasons of efficiency of 

DACOM Plant Plus recommendations. 

Conclutions. Results of one-year investigations confirmed possible effectiveness 

of DACOM Plant Plus decision support system in improvement of downy mildew 

control. Investigations are continued in 2009. 

15

Acknowledgements. Financial support from the Ministry of Agriculture of Latvia 

is gratefully acknowledged. 

Gauta 2009 06 22 


References 

1. Battilani P., Rossi V., Racca P., Giosue S. 1996. ONIMIL, a forecaster for primaru 

infection of downy mildew of onion. Bulletin OEPP/EPPO, 26: 567–576. 

2. Bashi E., Aylor D. E. 1985. Survival of detached sporangia of Peronospora destructor 

and Peronospora tabacina. Phytopathology, 73(8): 1135–1139. 

3. Bouma E. 2004. Decision support systems used in the Netherlands for reduction 

in the input of active substances in agriculture. EPPO Bulletin, 33(3): 461–466. 

4. Bulovienė V., Survilienė E. 2006. Effect of environmental conditions and inoculum 

concentration on sporulation of Peronospora destructor. Agronomy 

Research, 4: 147–150. 

5. Friedrich S., Leinhos G. M. E., Lopmeier F. -J. 2003. Development of ZWIREPO, 

a model forecasting sporulation and infection periods of onion downy mildew 

based on meteorological data. European Journal of Plant Pathology, 109: 

35–45. 

6. Gilles T., Phelps K., Clarkson J. P., Kennedy R. 2004. Development of 

MILIONCAST, an improved model for predicting downy mildew sporulation 

on onions. Plant disease, 88(7): 695–702. 

7. Hildebrand P. D., Sutton J. C. 1982. Weather variables in relation to and epidemic 

of onion downy mildew. Phytopathology, 72(2): 219–224. 

8. Palti J. 1989. Epidemiology, prediction and control of onion downy mildew caused 

by Peronospora destructor. Phytoparasitica, 17(1): 31–48. 

9. Peter T. N., Spencer-Phillips, Michael J., Jeger Y. 2004. Advances in downy 

mildew research. Kluwer Academic Publishers. 2: 81–89. 

10. Survilienė E., Valiuškaitė A., Raudonis L. 2008. The effect of fungicides on 

the development downy mildew of onions. Žemdirbystė – Agriculture, 95(3): 

171–179. 

11. Whiteman S. A., Beresford R. M. 1998. Evaluation of onion downy mildew 

disease risk in New Zealand using meteorological forecasting criteria. Proc. 51 st 

N. Z. Plant Protection Conference, 117–122. 

16


Svogūnų netikrosios miltligės integruotos kontrolės galimybės 

G. Bimsteine, L. Lepse, B. Bankina 

Santrauka 

Svogūnų netikroji miltligė, sukeliama Peronospora destructor, yra viena iš svarbiausių 

svogūnų ligų, nuo kurios būtina naudoti fungicidus. Šio tyrimo tikslas ir buvo surasti svogūnų 

netikrosios miltligės integruotos kontrolės būdus. Tyrimai atlikti Pūre sodininkystės 

tyrimų centre 2008 metais. Tyrime panaudoti trys svogūnų hibridai: ‘Safrane’ F 1 


ir ‘Alonso’ F 1 

. Tirti trys augalų apsaugos variantai: 1) fungicidai naudoti pagal „DACOM 

Plant Plus“ sistemą; 2) fungicidai naudoti pagal purškimo schemą; 3) fungicidai nenaudoti. 

Bandymuose naudoti fungicidai su veikliosiomis medžiagomis metalaksilu ir mankocebu. 

Pirmieji ligos simptomai pastebėti liepos 30 dieną. Skirtingiems hibridams liga vystėsi nevienodai. 

‘Alonso’ F 1 

netikrosios miltligės intensyvumas siekė 1,6 %, ‘Hypark’ F 1 

– 3,1 %, 

o ‘Safrane’ F 1 

– 4,5 %. Techninis fungicido naudojimo veiksmingumas priklausė nuo veislių 

ir purškimo varianto: purškiant pagal „DACOM Plant Plus“ sistemą, jis siekė 18,8–93,5 %, 

o schematinio purškimo atveju – 62,5–83,9 %. Veiksmingiausiai kelias ligai buvo užkirstas 

taikant DACOM programą. Siekiant išsamesnių rezultatų reikalingi tolimesni tyrimai. 

Reikšminiai žodžiai: fungicidai, Peronospora destructor, prognozės, purškimas. 

17




Interspecific relation peculiarities between soil and 

phytophatogenic fungi 

Danguolė Bridžiuvienė, Jūratė Repečkienė 

Institute of Botany, Žaliųjų ežerų st. 49, LT-08406 Vilnius, Lithuania, 

e-mail danguole.bridziuviene@botanika.lt 

Ecological methods of biological control of plant diseases became more and more popular 

in recent years. The aim of the investigation was to select soil fungi applicable for protection 

from causative agents of cultural plant diseases. Antagonistic activity of fungi was studied by 

pairing method using cultural blocs, cultural liquid and volatile metabolites. 43 fungal strains 

were examined against Alternaria brassicicola, A. radicina, A. tenuissima, Cladosporium 

cucumerinum, C. tenuissimum, Colletotrichum linicola, Fusarium culmorum, F. oxysporum, 

F. solani and Phoma sp. It was found that 36 of them showed varying degree of antagonistic 

activity against treated strains. Most strains suppressed the growth of Cladosporium genus 

phytopathogens and the growth of Fusarium genus strains was stopped rarely. Some of screened 

strains excreted fungicidal compounds to cultural liquid (for example, Trichoderma 

virens and Penicillium sp.) and some acted through their volatile metabolites (Arthrinium 

sphaerospermum and Trichoderma virens). The optimization of cultivation conditions as 

well as the choice of duration of cultivation is important factor to fungi ability to produce 

fungicidal compounds. 

Key words: fungi, fungicidal action, fungistatic action, plant pathogens. 

Introduction. Sustainable agriculture should apply environment-friendly strategies 

to avoid pollution of soil, water and products by chemicals. Consequently, 

in plant protection field the main attention has been concentrated on the biological 

control of plant pathogens in recent years. 

It is probable that biological control acts rarely through a single mechanism. 

Biological control agents may simply operate by occupying the ecological niche of 

the target organism so preventing its establishment and reducing its population. In 

this way antagonistic fungal species displace pathogens. Production of inhibitory to 

pathogens substances, including soluble, nonvolatile and volatile components, as well 

as extracellular enzymes are other ways of biological control. An inhibiting action of 

antagonists led to the reduction of phytopathogen conidia production and suppression 

of growth (Calistru et al., 1997; Schoeman et al., 1999; Kalko et al., 2001). 

Not all soils are rich in fungi with antagonistic abilities. It was found that introduction 

of several indigenous Trichoderma strains into the soil considerably affected 

19

the species composition. These changes influenced the phytosanitary state of soils 

and reduced the infectious lodging of conifer seedlings (Yakimenko, Grodnitskaya, 

2000). 

It was noticed that fungi isolated from the same environment as pathogens are 

more effective. For biological control the native strains should be used better and their 

population should be enriched in soil. Many studies of biocontrol agents have focused 

on Trichoderma species but the attempt to find more species with antagonistic abilities 

are made as well. Grapes inoculated with Botrytis cinerea were protected from disease 

by Trichoderma viride 81.2 % and by T. harzianum – 81.8 % (Machowicz-Stefaniak, 

1998). Active strains against Fusarium wilt diseases were found among Penicillium, 

Trichoderma, Gliocladium and Nigrospora genera fungi (Srinon et al., 2006). 

The study of interspecific relations between soil fungi and plant pathogens may 

turn a background for biocontrol technologies. The aim of the investigation was to 

screen soil fungal strains with antagonistic properties against plant pathogens. 

Object, methods and conditions. For primary antagonistic activity screening, 

43 fungal strains isolated from soil were used. As target fungi 10 phytopathogens 

were chosen: Alternaria brassicicola (Schwein.) Wiltshire 0722; A. radicina Meier, 

Drechsler et E. D. Eddy EŽ-25; A. tenuissima (Kunze) Wiltshire U-H-0; Cladosporium 

cucumerinum Ellis et Arthur 0721; C. tenuissimum Cooke 0679; Colletotrichum 

linicola Pethyrb. et Laff. U-L-07; Fusarium culmorum (W.G.Sm.) Sacc. 0691; 

F. oxysporum Schltdl. 0720; F. solani (Mart.) Appel et Wollenw. 0711 and Phoma sp. 

B-G-04. 

Tests for the biotic interaction among the fungi were carried out on malt agar in 

Petri dishes. The test fungi discs (1 cm in diameter) were cut from 7-day-old colonies 

and placed on malt agar medium, which was inoculated with phytopathogen spore suspension. 

Plates were incubated at the temperature of 26 °C. After 3 days of incubation, 

the diameter of fungicidal or fungistatic zones around agar discs was measured. The 

width of the resulting inhibitory zone was used as a measure of antagonistic activity 

(Билай, 1982). 

For detection of antagonistic effect of their culture filtrates, soil test-fungi were 

incubated in liquid Czapek medium for 7 days. Mycelium was removed by filtration. 

5-, 7- and 12-day culture filtrates (0.1 ml) were placed in hole made in agar inoculated 

with target-fungi. Plates without the filtrate served as control. Plates were incubated 

at the temperature of 26 °C. After 3 days of incubation the width of fungicidal or 

fungistatic zones around holes was measured (Билай, 1982). 

Detection of volatile antibiotics was performed by placing an inverted culture of 

a susceptible test-culture over that of the potential antagonist. Any reduction in the 

growth rate relative to control (expressed in %) by measuring colony diameter after 

4, 6 and 8 days was estimated. Some modified Czapek agar mediums were used for 

estimation of medium composition influence on volatile antibiotic production (Wheatly 

et al., 1997). The carbon source sucrose in standard medium was replaced by 2 % of 

glucose and in another variant in latter medium nitrogen source NaNO 3 

was replaced 

by 5 g of peptone. 

20

Results. Fungi isolated from various soils and plant remnants were selected for 

the primary screening of their antagonistic activity. In total 43 strains belonging to 15 

different genera were studied and 36 of them showed varying degree of antagonistic 

activity to phytopathogenic fungi belonging to genera Alternaria, Cladosporium, 

Colletotrichum, Fusarium and Phoma. The data of soil fungi interaction with various 

genera of phytopathogens are given in Table 1. 

Table 1. Number of soil fungal strains affecting the growth of phytopathogens 

from different genera 

1 lentelė. Tirtų dirvožemio mikromicetų padermių, veikiančių įvairių genčių fitopatogeninius 

mikromicetus, skaičius 

Genera of soil 

fungi 

Dirvožemio 

mikromicetai 

Number 

of tested 

strains 

Tirtų padermių 

skaičius 

Number of active strains against phytopathogens 

Aktyvių padermių skaičius 

Cladosporium Fusarium 

(2 strains (3 strains 

2 padermės) 3 padermės) 

Alternaria 

(2 strains 

2 padermės) 

21 

others 

(2 strains) 

kitos (2 padermės) 

Acremonium 6 2* (1)** 2 (1) (4) 1 (2) 

Arthrinium 5 (2) 2 (1) (5) (5) 

Chaetomium 4 4 3 2 (2) 4 

Cylindrocarpon 1 1 1 1 (1) (1) 

Clonostachys 1 0 0 0 1 

Cunninghamella 1 0 0 0 

Humicola 1 1 0 (1) (1) 

Mariannaea 1 1 1 1 2 

Mortierella 2 2 0 2 (2) 2 

Oidiodendron 1 1 2(1) (1) (2) 

Paecilomyces 3 1 0 2 (2) (2) 

Penicillium 8 4 (4) 7 (5) 6 (7) 6 (2) 

Talaromyces 2 1 (2) 2 (1) 1 (2) 2 (2) 

Trichoderma 5 2 (1) 4 (1) 2 (4) 2 (10 

Umbelopsis 2 (1) 1 (1) 1 (1) 1 (1) 

In total / Iš viso 43 21 (10) 27 (11) 18 (32) 20 (13) 

* – fungicidal effect / fungicidinis poveikis 

** – fungistatic effect / fungistatinis poveikis 

The biggest number of antagonists suppressed the growth of Cladosporium genus 

fungi – 27 strains (62.8 %) showed fungicidal activity and fungistatic activity was 

estimated mostly against Fusarium culmorum, F. oxysporum and F. solani (32 strains 

tested or 74.4 %). 

The greatest number of active strains was found among Chaetomium, Penicillium 

and Trichoderma species. It should be noted, that Chaetomium elatum Kunze 5OA; 

8OA and Ch. globosum Kunze EŽ-27 affected the phytopathogens not only by stopping 

their growth around the discs (due to extra-cellular metabolites), but by overgrowth 

of the target fungi as well (due to their competitive abilities). Therefore, the inhibitory 

zones around Chaetomium discs reached 5–8 mm in pairing with Alternaria, 

Fusarium, Phoma and even 6–11 mm in pairing with Colletotrichum, Cladosporium

genera strains. Among Penicillium the strains P. simplicissimum (Oudem.) Thom EŽ- 

25 and Penicillium sp. 0541 were more active as compared with other strains studied, 

especially against Alternaria and Cladosporium (diameter of sterile zones reached 

3–15 mm). Trichoderma virens (J. H. Mills, Giddens et A. A. Foster) Arx S29-1, 

T. hamatum (Bonord.) Bainier S37-1 and T. viride Pers S29-2 stopped the growth of 

the mentioned phytopathogens in 8–20 mm. 

Most active 18 strains were selected for further studies on ability to release 

antibiotics to nutrient medium. Four phytopathogens of different genus were chosen 

as target-fungi. The cultural filtrates of 15 strains affected fungistatically and/or fungicidally 

the growth of Alternaria radicina and Fusarium solani, 14 – Cladosporium 

cucumerinum and 12 – F. culmorum (Table 2). 

The obtained data showed that the inhibitory action of cultural filtrates differed in 

time ant specificity. The action of fungal cultural filtrates was mostly fungistatic and the 

width of growth suppression zone varied from 1 mm when, for example, Penicillium 

simplicissimum 5-day cultural filtrate was used against Fusarium culmorum, to 15 mm 

when Trichoderma virens S37-1 5-day cultural filtrate was used against Alternaria radicina. 

The growth of A. radicina most strongly was inhibited by 7-day cultural filtrate 

of Penicillium sp. 0541 (fungicidal zone 18 mm), Cladosporium cucumerinum – by 

7-day cultural filtrate of Gliocladium sp. Cr-3d (fungicidal zone – 8 mm), Fusarium 

culmorum – by 5-day cultural filtrate of Acremonium roseum (Oudem.) W. Gams 

U-5-60 (fungicidal zone 5 mm) and F. solani – by 5-day cultural filtrate of Penicillium sp. 

0541 (fungicidal zone 5 mm). 

Table 2. Antagonistic action of fungal cultural filtrates towards plant pathogen 

fungi (active strain number after 5, 7 and 12 cultivation days) 

2 lentelė. Mikromicetų kultūrinio skysčio antagonistinis poveikis fitopatogeniniams 

grybams (aktyvių padermių skaičius po 5, 7 ir 12 augimo parų) 

Phytopathogenic fungi 

Growth days 

Fitopatogeniniai grybai 

Auginimo paros Alternaria 

radicina 

Cladosporium 

cucumerinum 

Fusarium 

culmorum 

Fusarium 

solani 

5 4* (8)** 4 (9) 2 (5) 1 (6) 

7 6 (8) 8 (5) 0 (8) 3 (10) 

12 4 (8) 1 (8) 1 (8) 1 (12) 

* – fungicidal effect / fungicidinis poveikis 

** – fungistatic effect / fungistatinis poveikis 

Antibiotic effect of fungal cultural filtrates depended on cultivation timescale 

and changed in the course (Fig.). Some of them decreased gradually. The cultural 

filtrate of Chaetomium elatum 8OA showed the strongest fungicidal effect towards 

Alternaria radicina on 5 th cultivation day and made up 9 mm width inhibitory zone. 

On 7 th day it declined to 6 mm zone and on 12 th day reached only 3 mm. The latter 

antagonist showed towards Cladosporium cucumerinum a dual effect (fungistatic and 

fungicidal) at the same time. 

22

Fig. Dynamics of antagonistic activity of Chaetomium elatum 8OA 

culture filtrate on Alternaria radicina and Cladosporium cucumerinum 

Pav. Chaetomium elatum 8OA kultūros filtrato antagonistinio poveikio Alternaria radicina ir 

Cladosporium cucumerinum dinamika 

The action of Paecilomyces sp. S. p-20 cultural filtrate decreased as well, and 

even turned from fungicidal on 5 th day (5 mm inhibitory zone) to fungistatic on 

12 th day (5 mm suppression zone). The antagonistic (fungistatic) effect of Penicillium 

griseofulvum Dierckx 0457 towards Alternaria radicina, on the contrary, increased in 

the course of cultivation and made up 2 mm suppression zone on 5 th and later reached 

5 mm suppression zone. Mariannaea elegans G. Arnaud L14-3 has no antibiotic activity 

towards Alternaria radicina on 5 th cultivation day, but his cultural filtrate on 7 th 

day showed fungistatic effect (13 mm zone) and even fungicidal effect (7 mm zone) 

on 12 th day. Nevertheless, in most cases the highest antibiotic effect appeared on 7 th 

cultivation day. 

The action of volatile metabolites, produced by Arthrinium sphaerospermum 

Fuckel Pl-5/10, Mortierella alpina Peyronel 3OB, Penicillium simplicissimum EŽ-25 

and Trichoderma hamatum S37-1 towards the growth of plant pathogens was studied 

(Table 3). The most effective producer of volatile metabolites was T. hamatum S37-1. 

For example, the growth of Alternaria brassicicola was suppressed in 18–28 %, 

compared with control, (relative colony width was 82–72 %, respectively) under the 

action of Penicillium simplicissimum EŽ-25, but even in 73–86 % under the action 

of T. hamatum S37-1 (relative colony width was 27–14 %, respectively). The volatile 

metabolites of Arthrinium sphaerospermum Pl-5/10 effectively suppressed the growth 

of Alternaria brassicicola (relative size of colonies comparing to control after 8 days 

was 24 %), Colletotrichum linicola (55 %), Fusarium culmorum (62 %) and Phoma sp. 

(67 %). Mortierella alpina was more effective against Alternaria tenuissima, but 

stimulated the growth of some phytopathogenic strains (Cladosporium tenuissimum and 

Fusarium culmorum). The volatile metabolites of Penicillium simplicissimum EŽ-25 

were less effective (suppressed the growth of various strains 1–32 %). It was noticed 

that action of volatile metabolites was the greatest after 8 days growth, but in some 

cases it reached maximum just after 6 days (Arthrinium sphaerospermum Pl-5/10 and 

Mortierella alpina 3OB against Cladosporium and Fusarium genera strains). 

23

Table 3. Relative colony diameter (%) of phytopathogenic fungi under action of 

volatile metabolite produced on malt extract medium after 4, 6 and 8 cultivation 

days 

3 lentelė. Santykinis fitopatogeninių mikromicetų kolonijų, tirtų veikiant mikromicetų 

lakiaisiais metabolitais po 4, 6 ir 8 kultivavimo parų, skersmuo (%) 

Phytopathogenic fungi 

Fitopatogeniniai grybai 

Arthrinium 

sphaerospermum 

Pl-5/10 

Mortierella 

alpine 

3OB 

Penicillium Trichoderma 

simplicissimum hamatum 

EŽ-25 S37-1 

4 6 8 4 6 8 4 6 8 4 6 8 

Alternaria brassicicola 36 31 24 89 87 78 82 87 72 27 17 14 

Alternaria radicina 100 100 80 100 79 67 96 100 100 71 50 39 

Alternaria tenuissima 80 75 70 88 54 46 89 95 95 44 27 23 

Cladosporium cucumerinum 77 67 70 94 100 88 81 82 73 25 15 15 

Cladosporium tenuissimum 81 66 72 117 96 113 69 71 79 59 37 33 

Coletotrichum linicola 83 76 55 130 108 120 79 93 99 87 54 50 

Fusarium culmorum 80 75 62 120 125 138 88 81 75 92 75 63 

Fusarium oxysporum 92 89 100 99 89 100 90 88 81 56 50 50 

Fusarium solani 97 110 100 112 106 100 98 98 100 78 50 56 

Phoma sp. 90 78 67 125 121 128 82 80 68 75 61 51 

The cultivation of Arthrinium sphaerospermum Pl-5/10 and Mortierella alpina 

3OB on different media showed the volatile metabolite production dependence on 

medium composition (Table 4). 

Table 4. The effect of volatile metabolites of test-fungi Arthrinium sphaerospermum 

Pl-5/10 and Mortierella alpina 3OB grown on modified Czapek media 

towards plant pathogens (relative colony diameter after 7 cultivation days, %) 

4 lentelė. Mikromicetų Arthrinium sphaerospermum Pl-5/10 ir Mortierella alpina 3OB, 

augintų ant modifikuotų Čapeko terpių, lakiųjų metabolitų poveikis fitopatogenams (santykinis 

kolonijų skersmuo po 7 kultivavimo parų, %) 

Phytopathogenic 

fungi 

Fitopatogeniai 

grybai 

Arthrinium sphaerospermum Pl-5/10 

czapek czapek with czapek with 

with 2 % 2 % 2 % glucose 

glucose sucrose and peptone 

čapekas su čapekas su čapekas su 

2 % 2 % 2 % gliukozės 

gliukozės sacharozės ir peptonu 

czapek 

with 2 % 

glucose 

čapekas su 

2 % 

gliukozės 

Mortierella alpina 3OB 

czapek 

with 2 % 

sucrose 

čapekas su 

2 % 

sacharozės 

czapek with 

2 % glucose 

and peptone 

čapekas su 

2 % gliukozės 

ir peptonu 

1 2 3 4 5 6 7 

Alternaria 82 86 78 89 87 83 

brasicicola 

Alternaria 87 90 83 72 79 78 

radicina 

Alternaria 85 82 78 73 54 98 

tenuissima 

Cladosporium 

cucumerinum 

62 86 81 100 100 100 

24



1 2 3 4 5 6 7 

Cladosporium 70 81 78 91 96 63 

tenuissimum 

Colletotrichum 

86 82 80 88 108 84 

linicola 

Fusarium 78 83 68 98 125 78 

culmorum 

Fusarium 100 100 62 100 89 100 

oxysporum 

Fusarium 100 100 100 78 106 59 

solani 

Phoma sp. 76 076 56 57 121 77 

Glucose as a sole carbon source and peptone as nitrogen source (instead of 

NaNO 3 

) in modified Czapek medium favored the production of volatile metabolites 

of Arthrinium sphaerospermum Pl-5/10. The latter antagonist suppressed the growth 

of plant pathogens 17–38 % (the relative colony diameter was 83–62 %, except 

Fusarium solani). The volatile metabolites of A. sphaerospermum Pl-5/10 produced 

on Czapek medium with glucose suppressed most heavily the growth of Cladosporium 

genus pathogens and on Czapek with glucose and peptone – Phoma sp., Fusarium 

oxysporum and F. culmorum. 

Alike tendency was observed in Mortierella alpina 3OB case. Growing on 

Czapek with glucose and peptone the suppression of plant pathogens reached 16–41 % 

(except Cladosporium cucumerinum and Fusarium oxysporum). The best inhibition 

by M. alpina 3OB volatile metabolites on Czapek with glucose was noticed towards 

Phoma sp., on Czapek with sucrose – towards Alternaria tenuissima and on Czapek 

with glucose and peptone – towards Fusarium solani and Cladosporium tenuissimum. 

Furthermore this test-fungi growing on standard Czapek medium (with sucrose) stimulated 

the development of Colletotrichum linicola, Fusarium culmorum, F. solani 

and Phoma sp. 

Discussion. Species of Trichoderma and their biopreparations are mostly used 

against a number of fungal pathogens of agricultural importance (including the genera 

of Sclerotinia, Phytophthora and Botrytis) (Schoeman et al., 1999; Pięta et al., 

2003; Stankevičienė, Snieškienė, 2003). However, the data about antagonistic features 

of other typical soil fungi was presented (Chung Soohee, Kim Sang-Dal, 2005; 

Soytong et al., 2006). Consequently, it is supposed that a wide spectrum of fungi for 

various plant protections under certain conditions (in greenhouses, for field cultures 

and houseplants) could be used. It is difficult to find the strains active against all plant 

pathogens; therefore, the selection of active fungi against certain pathogens is very 

important. Our results of pairing tests on malt agar medium showed that fungal strains 

from Chaetomium, Mariannaea, Penicillium, Talaromyces and Trichoderma genera 

could inhibit the growth of all plant pathogens studied and other studied strains act 

selectively. For example, Mortierella genus strains suppressed Alternaria, Fusarium 

and Phoma but failed to affect the pathogens from Cladosporium genus. 

25

The tests with cultural filtrates and cultural discs of test fungi actions on plant 

pathogens showed different results. When Mortierella alpina 3OB discs were used they 

inhibited Fusarium solani growth, but its cultural filtrate showed no effect. The same 

tendency was established in case with Mariannaea elegans L14-3 and Trichoderma 

virens S29-1 cultural filtrates towards Alternaria radicina. These results confirm the 

idea of existence of different ways of interaction between fungal species. Some microbial 

antibiotics are nonvolatile and tend to diffuse through the liquid phase, in contrast 

to volatile metabolites, which spread as vapors (Schoeman et al., 1999). On the other 

hand, the same action of Penicillium sp. 0541 towards Fusarium solani or Alternaria 

radicina was noticed, notwithstanding what discs or cultural filtrate was used. These 

results coincide with A. M. Abdel-Sater (2001) data obtained, which showed the similar 

antagonistic activity in solid culture as well as of culture filtrates. 

The results obtained with inverted test-cultures over potential antagonist elucidated 

the strains that produce active volatile antibiotics. Such was Trichoderma hamatum 

S37-1, which inhibited the growth of Fusarium oxysporum even to 50 %, though its 

cultural filtrate showed only fungistatic action. 

Great dependence of antibiotic production on cultivation time and medium composition 

was estimated. It was found out that different fungal strains produce antibiotic 

metabolites most intensively not at the same time. For instance, cultural filtrate of 

Chaetomium elatum 8OA accumulated after 7 cultivation days and Paecilomyces sp. 

SP-20 stopped most powerfully Alternaria radicina after 3 days though after 12 days 

only fungistatic effect was fixed. 

Much data presented on direct correlation between degree of inhibition of fungi 

growth and nutrient medium composition (Engelkes et al., 1997; Rosenzwig, Stotzky, 

1980). Zh. I. Pavlovskaja et al. (1998) estimated that Trichoderma lignorum and 

Gliocladium catenulatum produce volatile metabolites against Fusarium sambucinum 

mostly on malt agar, whey and glucose. Our investigations with Arthrinium sphaerospermum 

Pl-5/10 and Mortierella alpina 3OB showed that the latter strains produce 

volatile metabolites mostly on modified Czapek medium with a sole carbon source 

glucose and nitrogen source – peptone. 

The obtained results suggest that some fungi might be promising as biocontrol 

agents. The great species inhibition selectivity and antibiotic production (non-volatile 

and volatile) dependence on cultivation conditions requires exhaustive scientific 

inquiry. The ecological significance shouldn’t be emitted and vegetative experiments 

and fungal complex action need to be studied, as well. 

Conclusions. 1. The ability to produce antibiotics was characteristic for 36 from 

43 tested fungal strains. Antifungal activity varied between different species isolates 

as well as between strains within the same species. 

2. The most active strains belonged to Chaetomium, Mariannaea, Penicillium, 

Talaromyces and Trichoderma genera. They mostly inhibited the growth of 

Cladosporium and Alternaria but were less effective against Fusarium. 

26

3. The action of nonvolatile and volatile metabolites and great dependence on cultivation 

conditions was noticed. Arthrinium sphaerospermum Pl-5/10 and Mortierella 

alpina 3OB strains produced volatile metabolites mostly on modified Czapek medium 

with a sole carbon source glucose and nitrogen source – peptone after 8 cultivation 

days. 

Gauta 2009 06 22 


References 

1. Abdel-Sater A. M. 2001. Antagonistic interaction between fungal pathogen 

and leaf surface fungi of onion (Allium cepa L.). Pakistan Journal of Biological 

Sciences, 4 (7): 838–842. 

2. Calistru C., McLean M., Berjak P. 1997. In vitro studies on the potential for biological 

control of Aspergillus flavus and Fusarium moniliforme by Trichoderma 

species – a study on the production of extracellular metabolites by Trichoderma 

species. Mycopathologia, 137(2): 115–124. 

3. Chung S., Kim S.-D. 2005. Biological control of phytopathogenic fungi by bacillus 

amyloliquefaciens 7079; suppression rates are better than popular chemical 

fungicides. Journal of microbiology and biotechnology, 15(5): 1 011–1 021. 

4. Engelkes C. A., Nuclo R. L., Fravel D. R. 1997. Effect of carbon, nitrogen and 

C : N ratio on growth, sporulation and biocontrol efficiency of Talaromyces flavus. 

Phytopathology, 5: 500–505. 

5. Kalko G. V., Vorobyov N. L., Lagutina T. M., Novikova I. I. 2001. Inhibition 

of the phytopathogenic fungus Fusarium oxysporum by microbe antagonists in 

peat. Mikologya i fitopatologya, 35(3): 68–75. 

6. Machowicz–Stefaniak Z. 1998. Antagonistic activity of epiphytic fungi from 

grape-vine against Botrytis cinerea Pers. Phytopathology Polonica, 16: 45–52. 

7. Pavlovskaja Zh. I., Mikhailova R. V., Lobanok A. G., Moroz I. V., Kobzarova V. S. 

1998. Antagonistic effect of Trichoderma lignorum (Tode) Harz OM 534 and 

Gliocladium catenulatum Gilman et Abbot 453 on Fusarium sambucinum 

Fuckel. Mikologya i fitopatologya, 32(3): 41–46. 

8. Pięta D., Pastucha A., Patkowska E. 2003. The use of antagonistic microorganisms 

in biological control of bean diseases. Horticulture and Vegetable growing, 

22(3): 401–405. 

9. Rosenzweig W. D., Stotzky G. 1980. Influence of Environmental factors on antagonism 

of fungi by bacteria in soil: nutrient levels. Applied and Environmental 

Microbiology, 39(2): 354–360. 

10. Schoeman M. W., Webber J. F., Dickinson D. J. 1999. The development of ideas 

in biological control applied to forest products. International Biodeterioration 

and Biodegradation, 43: 109–123. 

27

11. Soytong K., Srinon W., Rattanacherdchai K., Kanokmedhakul S., Kanokmedhakul 

K. 2006. Application of antagonistic fungi to control anthracnose disease 

of grape. Journal of Agricultural Technology, 1: 33–41. 

12. Srinon W., Chuncheen K., Jirattiwarutkul K., Soytong K., Kanokmedhaku S. 

2006. Efficacies of antagonistic fungi against Fusarium wilt disease of cucumber 

and tomato and the assay of its enzyme activity. Journal of Agricultural 

Technology, 2(2): 191–201. 

13. Stankevičienė A., Snieškienė V., 2003. Trichoderma viride against some of pink 

rot and wilt agents. Horticulture and Vegetable growing, 22(3): 395–399. 

14. Wheatly R., Hackett Ch., Bruce A., Kundzewicz A. 1997. Effect of substrate composition 

on production of volatile organic compounds from Trichoderma spp. inhibitory 

to wood decay fungi. International Biodeterioration and Biodegradation, 

39: 199–205. 

15. Yakimenko E. E., Grodnitskaya I. D. 2000. Effect of Trichoderma fungi on the 

soil micromycetes that cause infectious conifer seedling lodging in Siberian tree 

nurseries. Microbiology, 69(6): 850–854. 

16. Билай В. И. (ред.) 1982. Методы экспериментальной микологии. Науковa 

думка, Киев. 


Dirvožemio ir fitopatogeninių mikromicetų tarprūšinės sąveikos ypatumai 

D. Bridžiuvienė, J. Repečkienė 

Santrauka 

Augalų ligų ekologiški biologinės kontrolės metodai šiuo metu sulaukia vis daugiau dėmesio. 

Tyrimų tikslas buvo atrinkti dirvožemyje paplitusias mikromicetų padermes, pasižyminčias 

antagonistinėmis savybėmis prieš augalų ligų sukėlėjus. Antagonistinis mikromicetų aktyvumas 

buvo tirtas poravimo metodu naudojant kultūrų agaro blokus, kultūrinį skystį ir lakiuosius 

metabolitus. Tirtas 43 dirvožemio mikromicetų padermių aktyvumas Alternaria brassicicola, 

A. radicina, A. tenuissima, Cladosporium cucumerinum, C. tenuissimum, Colletotrichum lini, 

Fusarium culmorum, F. oxysporum, F. solani ir Phoma sp. atžvilgiu. Iš jų 36 padermės rodė 

didesnį ar mažesnį poveikį fitopatogeninėms grybų padermėms. Dauguma tirtų padermių 

stabdė Cladosporium genties fitopatogenų augimą, o Fusarium genties grybus dažniau veikė 

fungistatiškai. Kai kurios padermės (pvz., Trichoderma virens, Penicillium sp.) veikė išskirdamos 

antibiotines medžiagas į terpę, o kitos (pvz., Trichoderma virens ir Penicillium sp.) – lakiųjų 

metabolitų pagalba. Fungicidinių medžiagų gamybos intensyvumui turėjo įtakos mitybinės 

terpės sudėtis ir auginimo trukmė. Mikromicetų gebėjimui aktyviai gaminti fungicidinius 

junginius yra svarbus auginimo sąlygų optimizavimas bei kultivavimo trukmės parinkimas. 

Reikšminiai žodžiai: fitopatogenai, fungicidinis poveikis, fungistatinis poveikis, mikromicetai. 

28




Influence of boron fertilizer and meteorological 

conditions on red beet infection with scab and 

productivity 

Ona Bundinienė 

Lithuanian Institute of Horticulture, Kauno 30, LT-54333 Babtai, Kaunas distr., 

Lithuania, e-mail o.bundiniene@lsdi.lt 

Investigations of the additional red beet fertilization with boron fertilizers through 

leaves were carried out at the Lithuanian Institute of Horticulture, on Calc(ar)i-Epihypogleyc 

Luvisols – LVg-p-w-cc) of sandy loam on light loam in 2006–2007. There was little amount of 

humus and nitrogen in the soil, big amount of phosphorus, calcium and magnesium, average 

and big amount of potassium, average and big amount of boron; it was alkaline. There was 

investigated the influence of various boron fertilizers and meteorological conditions on scab 

prevalence in root-crops of different red beet cultivars and hybrids. 

In 2007 scab prevalence and disease intensity both in red beet hybrids and cultivars was 

2–3 times smaller than in 2006. The increase of temperature stimulated scab prevalence (for 

red beet ‘Boro’ F 1 

r = 0.88; for red beet ‘Kamuoliai 2’ r = 0.76) and increased intensity (correspondingly 

r = 0.88 and 0.85), and the increase of precipitation decreased scab prevalence 

and intensity (scab prevalence correspondingly r = -0.90 and -0.79, intensity r = -0.90 and 

-0.87). 

In both years of investigation root-crops of red beet cultivar ‘Kamuoliai 2’ were more 

infected by scab than red beet ‘Boro’ F 1 

. 

Boron fertilizers positively influenced the yield of red beet hybrids and cultivars and 

decreased scab prevalence and intensity (according to the average data of 2006–2007, on rootcrops 

of red beet cultivar ‘Boro’ F 1 

correspondingly 14.2 % and 15.0 %, on root-crops of red 

beet cultivar ‘Kamuoliai 2’ – 23.8 % and 6.3 %). Fertilizer Boramin Ca was the most effective 

to red beet. Economical efficiency of this fertilizer was correspondingly 2.6 % and 7.4 %. 

The increase of scab prevalence and intensity decreased red beet standard yield. The 

influence on root-crops of cultivar ‘Boro’ F 1 

was strong (correspondingly r = -0.91 and 

r = -0.95), and on root-crops of cultivar ‘Kamuoliai 2’ – average (correspondingly r = -0.44 

and r = -0.43). 

Key words: boron fertilizer, cultivar, hybrid, intensity, prevalence, productivity, red 

beet, scab, . 

29

Introduction. The most frequent nutrient deficiency encountered in cultivated 

beets is boron deficiency. Boron deficiency results in stunted plants, the deformation 

and death of the growing point, and slow growth (Nottingham, 2004, http://ourworld. 

compuserve.com/). When 10 t vegetable yield is grown, 23.7 kg of boron is obtained 

from the soil (Анспок, 1990). Boron deficiency occur in light-textured acid soils 

in humid regions because of boron tendency to leach, and in heavy-textured soils 

with high pH because boron is readily adsorbed under these conditions (Eisler, 2000; 

Gupta, 2007). The decrease of soil acidity creates conditions for bigger necessity of 

boron fertilizers, because soil boron agility decreases and it doesn’t get into the plants 

(Томсон, Троу, 1982). At high relative air humidity boron applied to the cotyledons 

was transported to hypocotyls and roots, whereas at low relative humidity no translocation 

of boron was detectable (Eichert, Goldback, 2006). There was no loss in yield 

of tops, roots or sugar from inadequate boron supply when soil boron exceeded 0.50 

mg B kg of soil (Christenson, Draycott, 2006). Boron fertilizers increase sugar beet 

yield 20.2 %, the amount of sugar – 0.3–0.6 % and the incidence of diseases of heart 

roots – 12.5–75.4 % (Анспок, 1990). 

Boron was found to reduce the severity of many diseases because of the function, 

which boron has on cell wall structure, plant membranes and plant metabolism 

(Dordas, 2008), and its deficiency caused some diseases (Томсон, Троу, 1982). Boron 

deficient beets had brown tops and roots were rough, scabby, and off colour (Gupta, 

Cutcliffe, 1985). 

Common scabies (Streptomyces scabies (Thaxter) Waksman and Henrici) are 

most seen on mature tuber or root-crop and consist of circular or angular lesions on the 

surface and rarely they may appear raised or penetrate to a few millimeters. Common 

scab gives an overall scruffy and unmarketable yield (Parry, 1990; Poljak et al., 2009; 

Koike et al., 2006). Streptomycetes are abundant in soils, rather than seed-borne and 

are controlled through agronomic rather than regulatory means by ensuring that soil 

moisture is maintained at field capacity by irrigating during the critical 4–6 week infection 

period and following tuber initiation (Elphinstone, 2007; Loria et al., 2006). The 

disease is seasonal and tends to most severe in light, freely drained alkaline soils after 

periods of dry summer weather (air temperature above 20 °C humidity – 50–70 %) 

and fertilization with fresh manure (Repšienė, Mineikienė, 2006; Ražukas et al., 2003; 

Elphinstone, 2007; Olanya et al., 2006; Parry, 1990; Waterer, 2002). Nutrient management 

and time of fertilization can decrease the severity of incidence of common scabies 

(Lambert et al., 2005; Davies et al.; Pavlista, 2005; Klikocka, 2009). 

T h e a i m o f t h e s t u d y was to investigate the influence of various boron 

fertilizers and meteorological conditions on scab prevalence and intensity in the rootcrop 

of various red beet cultivars and hybrids. 

Object, methods and conditions. Investigations were carried out at the Lithuanian 

Institute of Horticulture, on the calcaric epihypoghleyic luvisol of sandy loam on 

light loam (IDg8-k / Calc(ar)i- Epihypogleyc Luvisols – LVg-p-w-cc) in 2006–2007. 

Soil ploughing layer was 20–25 cm in thickness. There was little amount of humus 

(1.53–1.74 %) and nitrogen (38.2–60.5 kg ha -1 of soil) in the soil, very big amount 

of phosphorus (335–401 mg kg -1 of soil), calcium (6400–10850 mg kg -1 of soil) and 

30

magnesium (1 280–2 880 mg kg -1 of soil), average and big amount of potassium (191 

229 mg kg -1 of soil), average and big amount of boron (0.80–1.23 mg kg -1 of soil); it 

was alkaline (pH KCl 

7.2–7.6). 

Preplant – occupied fallow. Soil was cultivated and crop supervised according 

to the recommendations accepted at the LIH. There were grown red beet cultivars 

‘Boro’ F 1 

and ‘Kamuoliai 2’ on flat surface. Sowing scheme 62 + 8 cm. seed rate – 500 

thousand unit ha -1 of germinable seeds. 

Before red beet sowing, there was scattered N90P120K180 and additionally – N 30 

when red beet had 4–6 leaves. There was fertilized with ammonium saltpetre, granulated 

superphosphate and potassium magnesia. Boron fertilizers were applied for thee times: 

1) when red beet had 6–10 leaves; 2) after 12–14 days; 3) at root-crop formation, i. e. 

at the end of July (2006 – 06 21; 07 04; 07 17; 2007 06 27; 07 10; 07 19). For solution 

preparation 500 l ha -1 of water were used; its concentration was 0.2–0.3 %. 

S c h e m e o f t r i a l / Bandymo schema: 

Factor A / Veiksnys A – red beet hybrid and cultivar / burokėlių hibridas ir veislė: 

1. ‘Boro’ F 1 

2. ‘Kamuoliai 2’ 

F actor B / veiksnys B – different boron fertilizers / įvairios boro trąšos 

(B / F – background fertilization / foninis tręšimas N 90 + 30 

P 120 

K 180 

): 

1. Without boron / be boro. 

2. Boric acid / Boro rūgštis (17.2 % B). 

3. Tradebore / Tradeboras (15.4 % B). 

4. Tradebore Mo / Tradeboras Mo (15.4 % B and / ir Mo). 

5. Lyderis® Bor / Lyderis® Bor (10.5 % B, 0.01 % Zn, 0.007 % Cu, 

0.016 % Mn, 0.05 % Fe). 

6. Boramin Ca / Boramin Ca (6.5 % free amino acids / laisvųjų amino rūgščių, 

10.4 % CaO, 0.27 % B). 

Experiment was carried out in four replications. Fields lied out in random design. 

Record plot area – 4.8 m 2 . 

Red beet scabbiness was evaluated twice per vegetation: at the end of July and 

during harvest gathering. There were pulled up sex plants in each replication (24 plants 

per variant). After external inspection their root-crops injured by scab were evaluated 

in scores. The amount of injured root-crops (1), disease intensity (2) and economical 

efficiency of the applied means (3) was calculated in percents according to the 

formulas (Waller et al., 1998): 

P = n / N × 100, (1) 

here: P – injured root-crops (%), 

n – the number of scab injured root-crops, 

N – the number of inspected root-crops. 

R = Σ(a × b) × 100 / AK, (2) 

here: R – disease intensity (%), 

a – the number of root-crops injured in equal score, 

b – score of injury, 

A – the number of inspected root-crops, 

31

K – the highest score of injury (0–3), 

Σ – the sum of root-crops injured in equal score and 

multiplications of score values. 

Y = b – a / a × 100, (3) 

here: Y – economical efficiency of the applied means (%), 

a – yield obtained without applying the means, 

b – yield obtained without applying the means. 

Red beet yield was gathered when they reached technical maturity. 

M e t e o r o l o g i c a l c o n d i t i o n s. Thermal and irrigational conditions during 

red beet vegetation were characterized by monthly average air temperature, monthly 

precipitation sum, multiannual average and G. Selianinov hydrothermal coefficient – 

HTK (Table 1). 

HTK = Σp / 0.1 Σt, (4) 

here: Σt – period when the sum of active temperatures was bigger than 10° C; 

Σp – the sum of precipitation during the period, which average temperature 

> 10° C. 

When HTK ≥ 1.6 – excessive humidity, HTK 1–1.5 – optimal humidity, HTK 0.9– 

0.8 – slight draught, HTK 0.7–0.6 – average draught, HTK 0.5–0.4 – big draught, 

HTK ≤ 0.4 – very big draught (Bukantis, 2004, http://www. hkk.gf.vu.lt/). 

Meteorological conditions during the year of investigation were different: in 2006 

temperature of vegetation period 1.4 °C exceeded the multiannual average and was 

2.6 °C higher than in 2007, and in 2007 it was 1.2 °C lower than the average multiannual 

parameters (Table 1). In 2006 air temperature in May was alike multiannual one, 

and temperature parameters of all the other months exceeded the multiannual ones. 

In 2007 air temperature in all the vegetation months, except August, was lower than 

the average multiannual parameters (Table 1). 

Table 1. Meteorological conditions 

1 lentelė. Meteorologinės sąlygos 

Month 

Mėnuo 

Years 

Metai 

Long-term 

average 

Daugiamečiai 

vidurkiai 

The data of Kaunas meteorological station 

Kauno meteorologinės stoties duomenys 

Years 

Metai 

Long-term 

average 

Daugiamečiai 

vidurkiai 

Years 

Metai 

2006 2007 2006 2007 2006 2007 

Long-term 

average 

Daugiamečiai 

vidurkiai 

average air temperature precipitation sum 

vidutinė oro temperatūra (°C) kritulių suma (mm) 

HTK 

1 2 3 4 5 6 7 8 9 10 

Gegužė 12.6 11.2 12.3 74.0 104.4 50.7 1.89 2.23 1.33 

May 

Birželis 16.3 15.1 15.9 13.8 72.2 71.2 0.33 1.59 1.49 

June 

Liepa 

July 

19,3 15,2 17,3 30,2 173,6 75,3 0,50 3,68 1,40 

32



1 2 3 4 5 6 7 8 9 10 

Rugpjūtis 17.5 16.6 16.7 173.4 42.8 78.4 3.20 0.83 1.51 

August 

Rugsėjis 14.5 10.6 12.1 83.0 57.8 58.7 1.90 1.82 1.62 

September 

Spalis 9.7 5.4 7,1 47.0 51.2 50.5 1.69* - - 

October 

Average (sum) 15.0 12.4 13.6 70.2 83.7 64.1 1.56 2.03 1.47 

Vidutinė, suma 

* Note: In 2006 only first decade of October 

* Pastaba: 2006 metais tik pirmoji spalio dekada 

During both years of investigation precipitation rate exceeded the multiannual 

average, but rainy months and the amount of precipitation were different. In 2006 

June, July and October were dry, and in August, September and October of 2007 

precipitation rate was smaller than the multiannual average. The evaluation of the 

conditions during vegetation period according to the hydrothermal coefficient showed 

that agrometeorological conditions for red beet growing were as follows: in May and 

September of both the years of investigation – the excess of humidity, in June and 

July 2006 and in August 2007 – draught. Excessive humidity was in August 2006 and 

July 2007 (Table 1). 

Data significance was evaluated by the method of two-factorial dispersion 

analysis, using the program ANOVA, dependence – using the program STAT_ENG 

(Tarakanovas, Raudonius, 2003). 

Results. Meteorological conditions, cultivars of the grown red beet and fertilization 

with boron fertilizers influenced red beet root-crop infection by scab. In 2007 

scab prevalence and disease intensity in red beet hybrids and cultivars was 2–3 times 

smaller than these in 2006. Scab infection and disease intensity of red beet cultivar 

‘Kamuoliai 2’ in both years of investigation in July and in autumn at harvesting time 

was bigger than these of red beet cultivar ‘Boro’ F 1 

(Tables 2, 3). 

In 2006 at the end of July scab prevalence from boron fertilizers on root-crops 

of red beet cultivar ‘Boro’ F 1 

decreased 6.7 %, on cultivar ‘Kamuoliai 2’ – 32.5 %, in 

2007 – correspondingly 10.8 % amd 25.9 %. Disease intensity in red beet of cultivar 


on the average slightly increased, in ‘Kamuoliai 2’ – 13.4 % decreased, 

and in 2007 in red beet of hybrids and cultivars decreased correspondingly 12.5 % 

and 13.4 % (Table 2). The smallest scab prevalence and disease intensity in 2006, 

when the year was hot and less humid than 2007, on root-crops of red beet cultivar 


was after application of Lyderis®Bor, and on root-crops of red beet cultivar 

‘Kamuoliai 2’ – after application of boron fertilizers Boramin Ca. In cool and humid 

2007, Lyderis®Bor was the most effective both to hybrids and cultivars. 

Increasing temperature of June-July increased scab prevalence on red beet 

rootstocks (for red beet cultivar ‘Boro’ F 1 

r = 0.96; for red beet cultivar ‘Kamuoliai 

2’ r = 0.84) and intensity (correspondingly r = 0.95; r = 0.83). When precipitation 

rate increased, scab prevalence and intensity decreased (correspondingly prevalence 

r = -0.96 and r = -0.82, intensity r = -0.94 and r = -0.82). 

33

Table 2. Spread of common scabies on red beet root-crop in July 

2 lentelė. Paprastųjų rauplių plitimas ant burokėlių šakniavaisių liepos mėnesį 

Hybrid / cultivar 

(factor A) 

Hibridas / veislė 

(veiksnys A) 

Years 

Metai 

Fertilization with boron (factors B)* 

Tręšimas boru (veiksnys B) 

1 2 3 4 5 6 

34 

Babtai, 2006–2007 

LSD 05 

B 

R 05 

B 

Disease prevalence 

Ligos išplitimas (%) 


2006 79.2 79.2 79.2 66.7 62.5 75.0 12.1 

‘Kamuoliai 2’ 2006 95.8 87.5 62.5 70.8 54.2 41.7 


2007 25.0 29.25 12.5 20.8 0 8.3 8.2 

‘Kamuoliai 2’ 2007 41.7 29.2 8.4 25.0 0 16.7 

LSD 05 

A 2006 5.4 

R 05 

A 2007 3.7 

LSD 05 

A × B 

R 05 

A × B 

2006 18.0 

2007 12.14 

Disease intensity 

Ligos intensyvumas (%) 


2006 60.4 70.9 66.7 60.4 54.2 75.0 10.7 

‘Kamuoliai 2’ 2006 64.6 72.9 43.8 52.1 50.0 37.5 


2007 25.0 29.2 8.3 16.7 0 8.3 8.5 

‘Kamuoliai 2’ 2007 37.5 29.2 8.4 20.8 0 12.5 

LSD 05 

AB 2006 4.8 

R 05 

A 2007 3.8 

LSD 05 

A × B 

R 05 

A × B 

2006 15.9 

2007 12.6 

* Note: Variants of fertilization are shown in the methodical part 

* Pastaba: Tręšimo variantai pateikti metodinėje dalyje 

August of 2006 was hot and humid (HTK 3.2 – clear excess), and August of 2007 – 

cool and dry (HTK 0.83 – slight drought). After application of boron fertilizers, disease 

prevalence on root-crops of red beet cultivar ‘Boro’ F 1 

in 2006 decreased 17.5 %, intensity 

– 19.2 %, on root-crops of red beet cultivar ‘Kamuoliai 2’ correspondingly – 31.5 % 

and 7.5 %, in 2007 correspondingly 10.8 % and 13.1 %, 15.9 % and 5.0 % (Table 3). 

Therefore, in the year favourable to scab prevalence boron fertilizers were more effective 

in red beet cultivar ‘Kamuoliai 2’. The least scab prevalence and intensity on 

hybrid and cultivar root-crops in 2006 was observed applying fertilizer Boramin Ca, 

and in 2007 – on root-crops of cultivar ‘Boro’ F 1 

fertilizing with Lyderis®Bor and on 

root-crops of cultivar ‘Kamuoliai 2’ fertilizing with Tradebor Mo. 

When air temperature increased during vegetation period, scab prevalence (for 

red beet cultivar ‘Boro’ F 1 

r = 0.88; for red beet cultivar ‘Kamuoliai 2’ r = 0.76) and 

intensity (correspondingly r = 0.88 and r = 0.85) increased also. The increasing amount 

of precipitation, on the contrary, decreased scab prevalence and intensity (scab prevalence 

correspondingly r = -0.90 and r = -0.79; intensity correspondingly r = -0.90 

and -0.87).

Table 3. Spread of common scabies on red beet root-crop during harvesting 

3 lentelė. Paprastųjų rauplių paplitimas ant burokėlių šakniavaisių derliaus nuėmimo 

metu 

Hybrid / cultivar 

(factor A) 

Hibridas / veislė 

(veiksnys A) 

Years 

Metai 

Fertilization with boron (factors B)* 

Tręšimas boru (veiksnys B) 

1 2 3 4 5 6 

35 

Babtai, 2006–2007 

LSD 05 

B 

R 05 

B 

Disease prevalence 

Ligos išplitimas (%) 


2006 66.7 58.3 54.2 41.7 50.0 41.7 13.0 

‘Kamuoliai 2’ 2006 87.5 58.3 45.8 62.5 70.8 41.7 


2007 25.0 25.0 16.7 20.8 0 8.3 11.4 

‘Kamuoliai 2’ 2007 41.7 29.2 29.2 12.5 20.8 37.5 

LSD 05 

A 2006 5.8 

R 05 

A 2007 5.1 

LSD 05 

A × B 

R 05 

A × B 

2006 19.3 

2007 16.9 

Disease intensity 

Ligos intensyvumas (%) 


2006 66.7 45.8 54.2 45.8 50.0 41.7 11.9 

‘Kamuoliai 2’ 2006 56.3 41.7 43.8 56.3 64.6 37.5 


2007 25.0 25.0 16.7 20.8 0 8,3 9.1 

‘Kamuoliai 2’ 2007 26.4 22.9 29.2 12.5 16.0 26.4 

LSD 05 

A 2006 5.3 

R 05 

A 2007 4.1 

LSD 05 

A × B 

R 05 

A × B 

2006 17.7 

2007 13.5 

* Note: Variants of fertilization are shown in the methodical part 

* Pastaba: Tręšimo variantai pateikti metodinėje dalyje 

According to the average data of 2006–2007 investigations, additional fertilization 

through leaves with boron more influences the yield of red beet cultivar ‘Kamuoliai 2’. 

The standard red beet cultivar ‘Kamuoliai 2’ yield after fertilization increased on the 

average 1.1 t ha -1 (in 2006 – 0.3, in 2007 – 1.7 t ha -1 ), and this one of cultivar ‘Boro’ F 1 

increased only in 2007 (Fig.). 

Root-crop yield of red beet cultivar ‘Kamuoliai 2’ during the both years of investigation 

mostly (in 2006 – 1.9 t ha -1 , in 2007 – 3.1 t ha -1 ) increased after fertilization 

with Boramin Ca, in comparison with the yield of red beet grown without boron 

fertilizers. Economical efficiency of these boron fertilizers comprised correspondingly 

in 2006 4.9 %, in 2007 9.9 %; according to the average data of both years – 7.4 %. 

The yield of red beet cultivar ‘Boro’ F 1 

in 2006 mostly (only 0.4 t ha -1 ) increased after 

fertilization with Boramin Ca, and in 2007 – after fertilization with Boramin Ca and 

Lyderis®Bor (correspondingly 2.0 and 2.1 t ha -1 ). Economical efficiency of these fertilizers 

was correspondingly in 2006 1.2 %, in 2007 – 4.0 and 4.2 %; according to the

average data of both years 2.6 % (only Boramin Ca). Probably applying Boramin Ca, 

the main influence did calcium and free amino acids present in the composition of 

the fertilizer. 

Fig. Influence of boron fertilizers on standard yield of red beet 

Pav. Boro trąšų įtaka raudonųjų burokėlių standartiniam derliui 

The increase of scab prevalence and intensity decreased red beet standard yield. 

The influence on the yield of cultivar ‘Boro’ F 1 

was strong (correspondingly r = -0.91 

and r = -0.95), and on the yield of cultivar ‘Kamuoliai 2’ – average (correspondingly 

r = -0.44 and r = -0.43). 

Discussion. When it is warm (over 20 °C) and dry (humidity 50–70 %) during 

vegetation, the conditions for scab prevalence are very favourable (Ražukas et al., 

2003; Davies et al., 1976; Schoneveld, 1974). It was established in light loamy Albeluvisol 

of Western Lithuania that hydrothermal coefficient slightly influenced scab 

prevalence (Repšienė, Mineikienė, 2006). The data of our investigations carried out 

at the LIH showed that the temperature increase increased and precipitation increase 

– decreased scab prevalence and intensity. 

Potato resistance to common scab disease depends on genetic oneness and earliness 

of potato cultivars group (Ražukas ir kt., 2003; Klikocka, 2009). Trial results 

in Croatia showed that ‘Desire’ had significant higher percent of infected tubers with 

higher intensity of infection than ‘Courage’. In wet season, there was no evident on 

scabies infection and referred that lime addition increased scab incidence (Poljak 

et al., 2009). Scab lesions table beet and potato can cause considerable reductions in 

quality, resulting in loss of marketable yield (Koike et al., 2006; Ražukas et al., 2003). 

‘Avon Early’, ‘Elsoms’ 257 and ‘Bikor’ were the most resistant ‘globe’ beets, while 

‘New Globe’ and ‘Little Ball’ – most susceptible. ‘Boltardy’ and ‘Crimson Globe’, the 

cultivars widely grown in the fen, were among the more susceptible. The significance 

of this noticeable effect is not yet known, but it may reflect cultivar differences in the 

36

proportion of ‘root’ above and below ground (Lapwood et al., 1976). Our investigations 

showed that scab infection in root crops of red beet cultivar ‘Kamuoliai 2’ in both years 

of investigations was bigger than this one in red beet cultivar ‘Boro’ F 1 

. 

The application of fertilizers with S, Mg and B decreased the tuber infection rate 

and intensity of Streptomyces scabies, increasing potato tuber yield (Klikocka, 2009). 

According to the average data of 2006–2007, at the end of July boron fertilizers applied 

in our investigations 29.2 % decreased scab prevalence on root-crops of red beet cultivar 

‘Kamuoliai 2’, intensity – 18.3 %, in the autumn – 23.8 % and 6.3 %; on root-crops of 

red beet cultivar ‘Boro’ F 1 

– at the end of July correspondingly 8.8 % and 3.7 %, in the 

autumn – 14.2 and 15.0 %. Fertilizer Boramin Ca was the most effective to red beet. 

Economical efficiency of this fertilizer was correspondingly 2.6 % and 7.4 %. 

There was no effect of fertilizer material, application rate, or their interaction on the 

dry weight of alfalfa (Byers et al., 2001). Dried mass yield of sunflower was influenced 

by boron addition and dose 1.0 mg dm -3 provided the best yield and borax was more 

efficient (Marchetti et at., 2001). The data in the calcareous to a boron deficient soils of 

Pakistan showed that maximum sunflower biomass was produced with 1.0 mg kg -1 B, 

and application of ≥ 2.0 mg kg -1 proved toxic, resulting in drastic yield suppressions. 

Boron requirement can vary and depend on cultivar, plant age and plant part (Rashid, 

Rafique, 2005). A weekly foliar spray with boron (300 mg L -1 B) increased tomato 

marketable yield and fruit quality, reducing shoulder check incidence by 50 % compared 

to untreated plants (Huang, Snapp, 2009). In acid Oxisols of Brazil, boron application 

significantly increased common bean yield in only the first crop (Fageria et at., 2007). 

The foliar pulverization with boron brought benefit to the culture of beet, when boric 

acid or commercial product Supa bor® in concentration of 0.046 % was used (Saude 

et al., 2006, www.abhorticultura.com.). Boron and phosphorus did not affect yield of 

table beet, but boron application reduced incidence of root canker (Hemphill, 1982). 

Our investigations showed that boron fertilizers positively influenced red beet yield 

both of hybrids and cultivars. 

Conclusions. 1. The increase of temperature increased scab prevalence (for red 

beet ‘Boro’ F 1 

r = 0.88; for red beet ‘Kamuoliai 2’ r = 0.76) and intensity (correspondingly 

r = 0.88 and 0.85). The increase of precipitation decreased scab prevalence 

and intensity (scab prevalence correspondingly r = -0.90 and -0.79, intensity correspondingly 

r = -0.90 and -0.87). 

2. Scab infection and disease intensity in root crops of red beet cultivar 

‘Kamuoliai 2’ in both years of investigations was bigger than this one in red beet 

cultivar ‘Boro’ F 1 

. 

3. Boron fertilizers positively influenced red beet yield both of hybrids and cultivars. 

4. According to the average data of 2006–2007, at the end of July the applied 

boron fertilizers 23.8 % and 6.3 % decreased scab prevalence on root-crops of red beet 

cultivar ‘Kamuoliai 2’, on root-crops of red beet cultivar ‘Boro’ F 1 

– correspondingly 

14.2 and 15.0 %. 

37

5. Scab prevalence and intensity increase decreased red beet standard yield. The 

influence on the yield of cultivar ‘Boro’ F 1 

was strong (correspondingly r = -0.91 

and r = -0.95), and on the yield of cultivar ‘Kamuoliai 2’ – average (correspondingly 

r = -0.44 and r = -0.43). 

Gauta 2009 06 30 


References 

1. Byers D. E., Mikkelsen R. L., Cox F. R. 2001.Greenhouse evaluation of four 

boron fertilizer materials. Journal of plant nutrition, 24(4/5): 717–725. 

2. Bukantis A. 2004. Taikomoji meteorologija_2. Agrometeorologija. http://www. 

hkk.gf.vu.lt/ 

3. Christenson D. R., Draycott A. P. 2006. Nutrition – phosphorus, sulphur, potassium, 

sodium, calcium, magnesium and micronutrients – liming and nutrients 

deficiencies. In: A. P. Draycott (eds.), Sugar beet, Blackwell Publishing, Oxford, 

185–219. 

4. Davies J. R., McDole R. E., Callihan R. H. 1976. Fertilizer effects on common 

scab of potato and the relation of calcium and phosphate-phosphorus. 

Phytopathology, 66: 1 236–1 241. 

5. Dordas C. 2008. Role of nutrients in controlling plant diseases in sustainable 

agriculture. A review. Agronomy for Sustainable Development, 28: 33–46. 

6. Eichert T., Goldback H. E. 2006. Ambient air humidity affects phloem mobility 

of foliar-applied boron. Acta Hoticulturae, 700: 67–70. 

7. Eisler R. 2000. Handbook of chemical risk assessment. Metalloids, radiation, 

cumulative, index to chemicals and species, 3: 1 567–1 613. 

8. Elphinstone J. G. 2007. The canon of potato science: 11 bacterial pathogens. 

Potato Research, 50: 247–249. 

9. Fageria N. K., Baligar V. C., Zobel R. W. 2007. Yield, nutrient uptake, and soil 

chemical properties as influenced by liming and boron application in common 

bean in a no-tillage system. Communications in Soil Science and Plant Analysis, 

38(11&12): 1 637–1 653. 

10. Gupta U. C., Cutcliffe J. A. 1985. Boron nutrition of carrots and table beets 

grown in boron deficient soil. Communications in Soil Science and Plant 

Analysis, 16(5): 509–516. 

11. Gupta U. C. 2007. Boron. In: Handbook of Plant Nutrition. A. V. Barker, D. J. 

Pilbeam (eds.). CRC Press, 241–249. 

12. Hemphill D. D., Weber M. S., Jackson T. L. 1982. Table beet yield and boron 

deficiency as influenced by lime, nitrogen, and boron. Soil Science Society 

American Journal, 46: 1 190–1 192. 

38

13. Huang J., Snapp S. S. 2009. Potassium and boron nutrition enhance fruit quality 

in Midwest fresh market tomatoes. Communications in Soil Science and Plant 

Analysis, 40(11&12): 1 937–1 952. 

14. Klikocka H. 2009. Influence of NPK fertilization enriched with S, Mg, and 

micronutrients contained in Liquid fertilizer INSOL 7 on potato tubers yield 

(Solanom tuberosum L.) and infestation of tubers with Streptomyces Scabies and 

Rhizoctonia Solani. Journal of Elementology, 14(2): 271–288. 

15. Koike S. T., Gladders P., Paulus A. O. 2006. Vegetable diseases: color handbook. 

Academic Press. 

16. Lambert D. H., Powelson M. L., Stevenson W. R. 2005. Nutritional interactions 

influencing diseases of potato. American Journal of Potato Research, 82: 

309–319. 

17. Lapwood D. H., Adams M. J., Crisp A. F. 1976. The Susceptibility of Red Beet 

Cultivars to Streptomyces Scab. Plant Pathology, 25: 31–33. 

18. Loria R., Kers J. A., Joshi M. 2006. Evolution of plant pathogenicity in 

Streptomyces. Annual Review of Phytopathology, 44: 469–487. 

19. Marchetti M, E., Motomya W. R., Fabricio A. C, Noveino J. O. 2001. Sunflower, 

Heliantus annuus, response to sources and levels of boron. Maringa, 2(5): 

1 107–1 110. 

20. Nottingham S. 2004. Beetroot. http://ourworld.compuserve.com/homepages/ 

Stephen_Nottingham/ beetroot1.htm August 2004 SFN. 

21. Olanya O. M., Lambert D. H., Porter G. A. 2006. Effects of pests and soil management 

systems on potato diseases. American Journal of Potato research, 83(5): 

397–408. 

22. Parry D. W. 1990. Plant pathology in agriculture. New York, 385: 290–303. 

23. Pavlista A. D. 2005. Early-season applications of sulfur fertilizers increase potato 

yield and reduce tuber defects. Agronomy Journal, 97: 599–603. 

24. Poljak M., Horvat T., Nemet D., Buturac I. 2009. Effect of sustainable agricultural 

practices on control of potato scab in Croatia. Acta Horticulturae, 830(2): 

469–476. 

25. Rashid A., Rafique E. 2005. Internal boron requirement on young sunflower 

plants: proposed diagnostic criteria. Communications in Soil Science and Plant 

Analysis, 36(15&16): 2 113–2 119. 

26. Ražukas A., Čeponienė S., Repečkienė J. 2003. Potato cultivars resistance to common 

scab Streptomyces scabies. Sodininkystė ir daržininkystė, 22(3): 354–361. 

27. Repšienė R., Mineikienė E. V. 2006. Meteorologinių sąlygų ir skirtingų 

žemdirbystės sistemų įtaka bulvių ‘Mirta’ gumbų ligotumui bei derlingumui. 

Žemės ūkio mokslai, 3: 16–25. 

28. Saude I. B., Gracano D. B. C., Reis M. A., Cardoso V. P., Macieira G. A. A., da 

Fonseca F. H. A., Salgado P. J. A., Yuri J. E. 2006. Influence of boron sources 

and concentrations in the beet yield. //http. www.abhorticultura.com.br/.../trabalhos/ev_1/a344_t614_com. 

39

29. Tarakanovas P., Raudonius S. 2003. Agronominių tyrimų duomenų statistinė 

analizė, taikant kompiuterines programas ANOVA, STAT, SPLIT-PLOT iš paketo 

SELEKCIJA ir IRRISTAT. Akademija, Kėdainių r. 

30. Waller J. M., Ritchie B. J., Holderness M. 1998. IMI Technical Handbook No. 3: 

Plant Clinic Handbook. IMI, Bakeham Lane, Egham, Surrey TW20 9TY, UK, 

94. 

31. Waterer D. 2003. Impact of high soil pH on potato yields and grade losses to 

common scab. Canadian Journal Plant Science, 82: 583–586. 

32. Анспок П. И. 1990. Микроудобрения. Ленинград. 

33. Томсон Л. М., Троу Ф. Р. 1982. Почвы и их плодородие. Москва. 


Boro trąšų ir meteorologinių sąlygų įtaka burokėlių užsikrėtimui 

rauplėmis ir derlingumui 

O. Bundinienė 

Summary 

Burokėlių papildomo tręšimo boro trąšomis per lapus tyrimai atlikti Lietuvos sodininkystės 

ir daržininkystės institute 2006–2007 m. priesmėlio ant lengvo priemolio karbonatingajame 

sekliai glėjiškame išplautžemyje (IDg8-k / Calc(ar)i- Epihypogleyc Luvisols – LVg-p-w-cc). 

Dirvožemis buvo mažo humusingumo ir azotingumo, didelio fosforingumo, kalcingumo ir 

magningumo, vidutinio kalingumo ir kalingas, vidutinio ir didelio boringumo, šarmiškas. Tirta 

įvairių boro trąšų ir meteorologinių sąlygų įtaka rauplių išplitimui skirtingų burokėlių veislių 

ir hibridų šakniavaisiuose. 

2007 m. rauplių paplitimas ir ligos intensyvumas tiek hibrido, tiek veislės burokėliuose buvo 

2–3 kartus mažesnis negu 2006 m. Temperatūros didėjimas skatino rauplių plitimą (‘Boro’ F 1 

burokėliams r = 0,88; ‘Kamuoliai 2’ veislės burokėliams r = 0,76) ir stiprino intensyvumą 

(atitinkamai r = 0,88 ir 0,85), o kritulių kiekio didėjimas rauplių išplitimą ir intensyvumą mažino 

(išplitimas atitinkamai r = -0,90 ir -0,79, intensyvumas r = -0,90 ir -0,87). 

Abiem tyrimo metais ‘Kamuoliai 2’ veislės burokėlių šakniavaisiai buvo labiau užsikrėtę 

rauplėmis negu ‘Boro’ F 1 

burokėliai. 

Boro trąšos darė teigiamą įtaką tiek hibrido, tiek veislės burokėlių derliui ir mažino rauplių 

išplitimą bei intensyvumą (vidutiniais 2006–2007 metų duomenimis, ant ‘Boro’ F 1 

burokėlių 

šakniavaisių atitinkamai 14,2 ir 15,0 %, ant ‘Kamuoliai 2’ veislės burokėlių šakniavaisių – 23,8 % 

ir 6,3 %). Efektyviausios burokėliams buvo Boramin Ca trąšos. Ūkinis šių trąšų efektyvumas 

buvo atitinkamai 2,6 % ir 7,4 %. 

Rauplių išplitimo ir intensyvumo didėjimas mažino burokėlių standartinį derlių. ‘Boro’ F 1 

šakniavaisių derliui įtaka buvo stipri (atitinkamai r = -0,91 ir r = -0, 95), ‘Kamuoliai 2’ veislės 

burokėliams – vidutinė (atitinkamai r = - 0,44 ir r = - 0,43). 

Reikšminiai žodžiai: boro trąšos, derlingumas, hibridas, intensyvumas, išplitimas, 

raudonieji burokėliai, rauplės, veislė. 

40




Toxicity of insecticides to predatory mite 

Phytoseuilus persimilis in cucumber 

Laisvūnė Duchovskienė, Laimutis Raudonis, Rasa Karklelienė, 

Roma Starkutė 

Investigations were carried out with greenhouse cucumbers at the Lithuanian Institute of 

Horticulture in 2004–2005. The impact of three applied insecticides Envidor 240 SC (a. i. spirodiclofen 

240 g l -1 ) at the rates of 0.03 and 0.05 %, NeemAzal-T/S (a. i. 1 % azadirachtin A) 

at the rate of 0.5 % and Agri-50 (a. i. 28 % see weeds) at the rate of 0.3 % were determined 

on predatory mite Phytoseiulus persimilis. There were found significant differences between 

the number of predatory mites after application of Envidor 240 SC, NeemAzal-T/S and in 

untreated plants. Envidor 240 SC (0.05 % and 0.03 %) was from moderately toxic (mortality 

ranged from 50.98–66.67 %) 3 days after treatment to slightly toxic (25.0–48.75 % mortality) 

7 days after treatment. Envidor 240 SC was non-toxic 14 days after treatment. Agri-50 and 

NeemAzal-T/S were only slightly toxic (28.89–33.33 %) 3 days after treatment in 2005. Based 

on the results, NeemAzal-T/S and Agri-50 may be a useful part of Integrated Pest Management 

(IPM) programs. 

Key words: Agri-50, cucumber, Envidor, NeemAzal, Phytoseiulus persimilis. 

Introduction. Tetranychus urticae is one of the most serious pests in a number 

of agricultural systems. Its outbreaks are often a consequence of repeated and nonselective 

pesticide applications, which enhance pesticide resistance (Helle, Sabelis, 

1985). Resistance of T. urticae to numerous acaricides has developed difficulties controlling 

outbreaks of these pests (Carbonaro et al., 1986). Phytoseiulus persimilis can 

be affective as one of tools in the integrated pest management program for controlling 

T. urticae (Cote et al., 2002; Kim, Yoo, 2002). Despite successful suppression of 

T. urticae, limitations to the effectiveness of P. persimilis arise under certain conditions. 

The optimum conditions for rapid population development of P. persimilis are 

temperature of 27 °C and relative humidity (RH) of 60 % to 85 % (Stenseth, 1979). 

The effect of chemical classes like organophosphates, pyrethroids, organochlorines 

and carbamates on P. persimilis is from most to least harmful (Pratt, Croft, 2000). 

After threshold levels of T. urticae are surpassed, release of predators combined with 

compatible acaricide is more effective than using chemical or biological control tactics 

alone (Trumble, Morse, 1993). 

Object, methods and conditions. Investigations were carried out with greenhouse 

cucumbers at the Lithuanian Institute of Horticulture in 2004–2005. The general 

EPPO standards were used for investigations (Anon, 1997). 

Envidor 240 SC (a. i. spirodiclofen 240 g l -1 ) at the rates of 0.03 % and 0.05 %, 

41

NeemAzal-T/S (a. i. 1 % azadirachtin A) at the rate of 0.5 % and algae extracts Agri-50 

(a. i. 28 % see weeds) at the rate of 0.3 % water spraying solution was applied. The 

plot size for tests was 5 m 2 . Four replications were made in a randomized block design. 

20 leaves per plot were observed to determine the number of P. persimilis for evaluation 

the efficiency of tested products. Phytoseiulus persimilis adults were obtained 

from Biobest (Belgium). Six predators per square meter were released after arrival. 

The cucumber plants were naturally infected by Tetranychus urticae. Insecticide was 

applied 10 days after P. persimilis introduction. Predators were recorded as follows: 

24 h before and 3, 7 and 14 days after treatment. The counts of mortality of adults 

and larvae were corrected by Abbott’s formula (1925). We used quantitative toxicity 

categories from International Organization for Biological Control for assessment of 

pesticide toxicity to predatory and phytophagous mites in field trials: nontoxic (< 25 % 

mortality), slightly toxic (25–50 %), moderately toxic (51–75 %), very toxic (> 75 %) 

(Hassan et al., 1985). 

The number of predatory mites among treatments was compared with a single 

factor analysis of variance (ANOVA). Specific differences were identified with 

Duncan’s multiple range test. 

Results. Predatory mites were numerous in 2004 comparing with 2005. Decrease 

of P. persimilis number from exposure to residues of Envidor 240 SC was mostly 

significantly greater as observed in control comparing with NeemAzal-T/S and 

Agri-50 (especially Envidor 240 SC at the rate of 0.05 %) after application (Table 

1). 

Table 1. Number of predatory mites Phytoseiulus persimilis after application of 

insecticides 

1 lentelė. Grobuoniškų erkių Phytoseiulus persimilis gausumas po purškimo insekticidais 

Babtai, 2004–2005 

Mean number of Phytoseiulus persimilis, unit/ leaf 

Vidutinis Phytoseiulus persimilis skaičius ant lapo 

Treatments 

before 

days after application 

Variantai 

application 

dienos po purškimo 

prieš purškimą 3 7 14 

1 2 3 4 5 

2004 

Untreated 

4.4 ab 5.1 c 5.6 b 3.2 a 

Nepurkšta 

Envidor 240 SC 0.05 % 5.6 ab 2.3 a 4.2 ab 2.7 a 

Envidor 240 SC 0.03 % 5.2 ab 2.5 a 3.8 ab 3.1 a 

NeemAzal-T/S 1 % EC 0.5 % 4.2 ab 4.1 abc 5.4 ab 6.2 b 

AGRI-50 0.3 % 5.7 b 4.8 bc 5.0 ab 4.1 a 

42



1 2 3 4 5 

2005 

Untreated 

3.0 abc 4.5 c 8.0 c 3.4 abc 

Nepurkšta 

Envidor 240 SC 0.05 % 2.0 a 1.5 a 4.1 a 2.6 a 

Envidor 240 SC 0.03 % 2.5 abc 2.0 ab 4.7 a 3.3 abc 

NeemAzal-T/S 1 % EC 0.5 % 3.2 bc 3.0 bcd 7.2 bc 4.4 bc 

AGRI-50 0.3 % 3.5 c 3.2 bcd 6.3 abc 4.8 c 

Note: Means followed by the same letter are not different significantly (P = 0.05) according to 

Duncan’s multiple range test 

Pastaba: Reikšmės, pažymėtos tomis pačiomis raidėmis, pagal Dunkano kriterijų (P = 0,05) iš 

esmės nesiskiria. 

There were not found statistical differences of number of predatory mites treated 

with Envidor 240 SC 0.03 and 0.05 %. The number of P. persimilis treated with Agri-50 

did not differ significantly comparing with NeemAzal-T/S. Decrease of P. persimilis 

was insignificant after treatment with Agri-50, NeemAzal-T/S and in untreated plots. 

Small decrease of predatory mites comparing with untreated plots was observed only 

3 days after spraying. A number of mites began to increase after 7 days, but after 14 

days in all the treatments number of P. persimilis decreased. 

Envidor 240 SC (0.05 % and 0.03 %) was from moderately toxic (mortality ranged 

from 50.98–66.67 %) to slightly toxic (25.0–48.75 % mortality) (Table 2). 

Table 2. Residue toxicity to Phytoseiulus persimilis after application of insecticides 

2 lentelė. Išliekamasis toksiškumas Phytoseiulus persimilis po purškimo insekticidais 

Treaments 

Variantai 

43 

Babtai, 2004–2005 

Mortality of Phytoseiulus persimilis 

Phytoseiulus persimilis žuvimas (%) 

after application 

po purškimo 

3 d. 7 d. 14 d. 

2004 2005 2004 2005 2004 2005 

Envidor 240 SC 0.05 54.90 66.67 25.00 48.75 15.62 23.53 

Envidor 240 SC 0.03 50.98 55.55 32.14 41.25 3.12 2.94 

NeemAzal-T/S 1% EC 0.5% 19.61 33.33 3.57 10.00 -93.75 -29.41 

AGRI-50 5.88 28.89 10.71 21.25 -28.75 -41.18 

Envidor 240 SC was non-toxic only 14 days after treatment. Three days after spraying 

Agri-50 and NeemAzal-T/S were only slightly toxic (28.89–33.33 %) in 2005. 

Discussion. Numerous field studies using P. persimilis alone and P. persimilis 

combined with compatible acaricides demonstrate that adequate control can be 

achieved with correct combination of acaricides and this predatory mite (Osborne,

Patitt, 1985, Cashion et al., 1994). The level of compatibility will usually depend on 

the post application interval (Cote et al., 2002). We measured toxicity of acaricides 

to commercially available P. persimilis. We did not consider side effects, which can 

occur from acaricide residues like other authors (Oomen et al., 1991). 

Decrease of P. persimilis number from exposure to residues of Envidor 240 SC 

was significantly greater than this observed in untreated plots 3 days and 7 days after 

spraying in 2005. There were not found statistical differences of number of predatory 

mites treated with different rates of Envidor 240 SC and mortality of P. persimilis 

was similar. The toxic effect of pesticides on mites depends on the chemistry of pesticides, 

their rates, microclimatic conditions and development stages of mites (Pratt 

and Croft, 2000; Papaioannou-Souliotis et al., 2000; Stenseth, 1979). It was found that 

Envidor 240 SC was non-toxic to predatory mites in strawberry under field conditions 

(Raudonis, 2006). In our treatment Envidor 240 SC (0.05 and 0.03 %) was from 

moderately toxic (after 3 days) to slightly toxic (after 7 days) to P. persimilis. The 

mortality ranged from 25.0–66.67 %. Envidor 240 SC (both rates) was non-toxic and 

had not long-lasting effect 14 days after spraying. 

It was found that Neem products are active for only a short period, but all neem 

products may not be equally compatible with P. persimilis (Cote et al., 2002). Direct 

application of neem formulation containing 80 % neem oil at a rate of 3 % was highly 

toxic to P. persimilis (Papaioannou-Souliotis et al., 2000). NeemAzal-T/S containing 

triterpenoid azadirachtin has a number of useful properties for insect control due to its 

repellency, feeding and oviposition deterrence, insect growth regulator. This product 

is considered as safe to the environment and reduces the rate of pest development 

(Schmutterer, 1990; Pavela, Holy, 2003). On the other hand, Azadirachtin was the 

least toxic from tested insecticides to other predatory mite Neoseiulus californicus 

(Castagnoli et al., 2005). Nevertheless, in our treatment NeemAzal-T/S was only 

slightly toxic 3 days after treatment. Neem products may be a useful part of Integrated 

Pest Management (IPM) programs, however, its short residual toxicity may not suppress 

large population of two spotted spider mite (Cote et al., 2002). The number of 

P. persimilis on leaves treated with Agri-50 did not essentially differ comparing with 

NeemAzal-T/S and both products did not statistically reduce the number of P. persimilis 

comparing with unsprayed plots. Proper insecticide selection may create favorable 

conditions for combination predator release and the use of chemical products. 

Conclusions. Envidor 240 SC (0.05 and 0.03 %) was moderately toxic (mortality 

of P. persimilis ranged from 50.98 to 66.67 %) three days after treatment and slightly 

toxic (mortality 25.0–48.75 %) seven days after treatment. Only fourteen days after 

treatment Envidor 240 SC was non-toxic. 

NeemAzal-T/S and Agri-50 were slightly toxic (mortality of P. persimilis ranged 

from 28.89 to 33.33 %) only three days after treatment. Based on the results, NeemAzal- 

T/S and Agri-50 could be a used for Integrated Pest Management (IPM) programs. 

Gauta 2009 06 30 


44

References 

1. Abbott W. S. 1925. A method for computing the effectiveness of an insecticide. 

Journal of Economic Entomology, 18: 265–267. 

2. Anon. EPPO Standards. 1997. Guidelines for the efficacy evaluation of plant protection 

products. Insecticides & Acaricides. Editor European and Mediterranean 

Pl. Prot. Org. Paris, 3: 231. 

3. Carbonaro M. A., Moreland D. E., Edg V. E., Matoyama N., Rock G. C., 

Dauterman W.C. 1986. Studies of the mechanisms of cyhexatin resistance in the 

two-spotted spider mite Tetranychus urticae (Acari: Tetranychidae). Journal of 

Economic Entomology, 79: 579–580. 

4. Cashion G. J., Bixler H., Price J. F. 1994. Nursery IPM trials using predatory 

mites. Proceedings of the Florida State Horticultural Society, 107: 220–222. 

5. Cote K. W., Lewis E. E., Schultz P. 2002. Compatibility of acaricide residues with 

Phytoseiulus persimilis and their effect on Tetranychus urticae. HortScience, 

37(6): 906–909. 

6. Castagnoli M., Liguori M., Simoni S., Duco C. 2005. Toxicity of some insecticides 

to Tetranychus urticae, Neoseiulus californicus and Tydeus californicus. 

BioControl, 50: 611–622. 

7. Hassan E., Oomen, P. A., Overmeer W. P. J., Plevoets P., Reboulet J. N., 

Rieckmann W., Samsoe-Petersen L., Shires S. W., Staubli A., Stevensen J., 

Tuset J. J., Vanwetswinkel G., Zon A. Q. 1985. Standard methods to the test of 

side effects of pesticides on natural enemies of insects and mites developed by 

the IOBC/WPRS. Bulletin OEPP/EPPO, 15: 214–255. 

8. Helle W., Sabelis M. W. 1985. Spider Mites. Their Biology, natural Enemies and 

Control. Elsevier, Amsterdam, Oxford, New York, Tokio, 1 A: 391–395. 

9. Kim S. S., Yoo S. S. 2002. Comparative toxicity of some acaricides to the predatory 

mite, Phytoseiulus persimilis and the two-spotted spider mite Tetranychus 

urticae. BioControl, 47 (5): 563–573. 

10. Oomen P. A., Romeijn G., Wiegers G. L. 1991. Side effects of 100 acaricides on 

the predatory mite Phytoseiulus persimilis, collected and evaluated according 

to the EPPO guideline. Bulletin European and Mediterranean Plant Protection 

organization (OEPP/EPPO), 21: 701–712. 

11. Osborne L. S., Petitt F. L. 1985. Insecticidal soap and the predatory mite 

Phytoseiulus persimilis (Acari: Phytoseiidae), used in management of the twospotted 

spider mite (Acari: Tetranychidae) on greenhouse grown foliage plants. 


12. Papaioannou-Souliotis P., Markouiannaki-Printziou D., Zoaki-Malissiova D. 

2000. Side effects of Neemark (Azadirachta indica A. Juss) and two new vegetable 

oil formulations on Tetranychus urticae Koch. and its predator Phytoseiulus 

persimilis Athias-Henriot. Bollettino di Zoologia Agraria e di Bachicoltura, 

32: 25. 

45

13. Pavela R., Holy K. 2003. Effect of azadirachtin on larvae of Lymantria dispar, 

Spodoptera litoralis and Mamestra brassicae. Sodininkystė ir daržininkystė, 

22(3): 434–441. 

14. Pratt P. D., Croft B. A. 2000. Toxicity of pesticides registered for use in landscape 

nurseries to the acarine biological control agent, Neoseiulus fallacis. Journal 

of Environmental Horticulture, 18: 197–201. 

15. Raudonis L. 2006. Comparative toxicity of spirodiclofen and lambdacihalotrin 

to Tetranychus urticae, Tarsonemus pallidus and predatory mite Amblyseius 

andersoni in a strawberry site under field conditions. Agronomy Research, 4: 

317–326. 

16. Schmutterer H. 1990. Properties and potential of natural pesticides from neem 

tree, Azadirachta indica. Annual Review of Entomology, 35: 271–297. 

17. Stenseth C. 1979. Effect of temperature and humidity on the development of 

Phytoseiulus persimilis and its ability to regulate populations of Tetranychus 

urticae (Acarina: Phytoseiidae, Tetrancyhidae). Entomophaga, 249: 311–317. 

18. Trumble J. T., Morse J. P. 1993. Economics of integrating the predaceous mite 

Phytoseuilus persimilis (Acarina: Phytoseiidae) with pesticides in strawberries. 



Insekticidų toksiškumas grobuoniškoms erkėms 

Phytoseuilus persimilis agurkuose 

L. Duchovskienė, L. Raudonis, R. Karklelienė, R. Starkutė 

Santrauka 

Tyrimai atlikti Lietuvos sodininkystės ir daržininkystės institute 2004–2005 metais šiltnamyje 

augintuose agurkuose. Tirtas trijų insekticidų: Envidor 240 SC (v. m. spirodiklofenas 240 g l -1 ) 

0,03 ir 0,05 %, Nimazal-T/S (v. m. 1% azadirachtinas A) 0,5 % ir Agri-50 (v. m. 28 % jūros žolė) 

0,3 % poveikis grobuoniškoms erkėms Phytoseiulus persimilis. Buvo rasti esminiai skirtumai 

tarp grobuoniškų erkių skaičiaus po purškimo Envidor 240 SC, Nimazal T/S ir nepurkštų augalų. 

Envidor 240 SC (0,05 % ir 0,03 %) buvo nuo vidutiniškai toksiško (P. persimilis mirtingumas 

kito nuo 50,98 iki 66,67 %) praėjus 3 dienoms po purškimo iki silpnai toksiško (P. persimilis 

mirtingumas kito nuo 25,0 iki 48,75 %) praėjus 7 dienoms po purškimo. Envidor 240 SC (0,05 % 

ir 0,03 %) po 14 dienų buvo netoksiškas. Agri-50 ir Nimazal T/S 2005 metais buvo silpnai 

toksiški (28,89–33,33 %) praėjus 3 dienoms po purškimo. Remiantis tyrimų rezultatais, Nimazal- 

T/S ir Agri-50 gali būti sėkmingai naudojami integruotos augalų apsaugos (IAA) programose. 

Reikšminiai žodžiai: Agri-50, agurkai, Envidor, Nimazal-T/S, Phytoseiulus persimilis. 

46




Effect of Abamectin on two-spotted spider mite and 

leaf miner flies in greenhouse cucumbers 

Laisvūnė Duchovskienė, Elena Survilienė 


Lithuania, e-mail l.raudonis@lsdi.lt 

Effect of Abamectin 18 g l -1 on two-spotted spider mite (Tetranychus urticae Koch.) and 

leaf miner flies (Liriomyza strigata, Liriomyza bryoniae) was studied in greenhouse cucumbers 

in 2005–2006. Efficiency of Abamectin 18 g l -1 0.12 % and 0.1 % against two-spotted spider 

mite was respectively 92.24–100 % and 82.8–99.2 %. Efficiency of insecticide Abamectin 

18 g l -1 0.075 % and 0.05 % against two-spotted spider mite was respectively 81.0–100 % 

and 72.4–94.0 %. Efficiency of Abamectin 18 g l -1 0.12 % and 0.1 % against leaf miner flies 

was respectively 63.6–80.3 % and 60.4–75.26 %. Efficiency of insecticide Abamectin 18 g l -1 

0.075 % and 0.05 % against leaf miner flies was respectively 35.9–54.4 % and 30.7–49.6 %. 

Azadirachtin A 10 g l -1 and Spirodiclofen 240 g l -1 were less effective against two-spotted 

spider mite and leaf miner flies. 

Key words: Abamectin, Azadirachtin A, cucumber, Liriomyza bryoniae, Liriomyza 

strigata, Spirodiclofen, Tetranychus urticae. 

Introduction. Tetranychus urticae is major pest on field crops, glasshouse crops, 

horticultural crops, ornamentals and fruit trees (Van de Vrie et al., 1972). It is one of 

the most serious pests in a number of agricultural systems; its outbreaks are often 

a consequence of repeated and non-selective pesticide applications, which enhance 

pesticide resistance (Helle, Sabelis, 1985). However, spider mite does not attack all 

plants to the same degree because of differences in nutritive and toxic constituents 

(Boom et al., 2003). Moreover, it can easily adapt to plant varieties that have been 

especially selected for resistance, as was shown for e. g. in tomato, broccoli and 

cucumber (Fry, 1989). Two-spotted spider mite feeds on leaves causing damage in 

chlorophyll and producing white spots that may become more or less coherent with 

time (Nachman, Zemek, 2002). Leafmining flies Liriomyza strigata, Liriomyza bryoniae 

are frequent species in Lithuanian glasshouses and around them (Ostrauskas 

et al., 2003, Ostrauskas et al., 2005). Adult leaf miner flies puncture plant leaves for 

feeding and oviposition. Larvae reduce the photosynthetic activity of the leaves by 

mining (Parrella et al., 1985). The decreased leaf productivity by T. urticae and leaf 

miner fly larvae feeding caused biomass reduction and altered the pattern of dry matter 

partitioning in the plant; damaged plants accumulated more dry matter in leaves, 

47

and partitioning of dry matter to fruits was hindered (Park, Lee, 2005; Parrella et al., 

1985). 

Thus experiments performed in 2005–2006 were designed to clarify how active ingredient 

Abamectin affect two-spotted spider mite and leaf miner flies in cucumber. 

Object, methods and conditions. The investigations were carried out in greenhouse 

cucumber during growing seasons of 2005–2006 in Lithuania as efficiency trials 

of Abamectin 18 g l -1 against two-spotted mite and leaf miner flies. Greenhouse was 

covered with double film, the sides of greenhouse – with plastic. The general principles 

of investigations were in accord with EPPO standards (Anon, 1997). 

The trial was carried out according to trial plan as follows (Table 1). 

Table 1. Trial plan 

1 lentelė. Bandymo planas 

Treatment 

Variantas 

Untreated 

Nepurkšta 

Abamectin 18 g l -1 



Abamectin 18 g l-1 

Azadirachtin A 10 g l -1 

(standard / standartas) 

Spirodiclofen 240 g l -1 


Trade name 

Registruoto preparato pavadinimas 

Vertimex 1.8 EC 




NeemAzal-T/S 

Envidor 240 SC 

Rate l per ha 

Norma l/ha (%) 

1.2 (0.12 %) 

1.0 (0.1 %) 

0.75 (0.075 %) 

0.5 (0.05 %) 

5.0 (0.5 %) 

0.5 (0.05 %) 

The size of plot for tested plants was 10 m 2 . Four replications were made in randomized 

block design. To determine efficiency of the examined insecticide 10 plants 

per plot were observed. After first observation leaves with leaf miner flies mines was 

removed. Mines were identified according to Pakalniškis et al. (2005). Pests were 

recorded as follows: 24 h before and 3, 7, 9 and 14 days after treatment. Biological 

efficiency of insecticides in pest control was presented as percentage of pest mortality. 

Statistical analysis of results was done. 

Microclimate conditions in greenhouses at the time of treatments were favourable 

for insecticide applications (Table 2). 

Efficacy of abemectin against two-spotted spider mites was calculated: x = 100 

(1 – A b/B a) (x – efficacy of insecticide, %, A – number of mites per leaf before spraying 

in untreated plot, B – number of mites per leaf before spraying in treated plot, 

a – number of mites per leaf after spraying in untreated plot, b – number of mites per 

leaf after spraying in treated plot). 

The number of two-spotted spider mites and damaged leaves of leaf miner flies 

was compared among treatments in this study with single factor analysis of variance 

(ANOVA). Specific differences were identified with Duncan’s multiple range test. 

48

Table 2. Circumstances during application 

2 lentelė. Sąlygos purškimo metu 

Date of application 

Purškimo data 

2005-07-20 2006-06-23 

Code of plant growth stage during application (BBCH scale) 

Augalo augimo tarpsnis (pagal BBCH skalę)* 

Temperature on the date of application 

Temperatūra (°C) 

Relative air humidity on the date of application 

73–74 

18 

81 

51–52 

18 

80 

Santykinė oro drėgmė, % 

* Growth stages are described according to BBCH scale 

* Augalų augimo tarpsniai apibūdinami pagal BBCH skalę (Meier, 1997) 

Results. Trial data shows that insecticide Abamectin 18 g l -1 , at rates of 0.12, 

0.10, 0.075 % spraying solution was highly effective against two-spotted spider mites 

in 2005. There were not found any statistical differences of number of two-spotted 

spider mites between Abamectin 18 g l -1 0.12 % and Abamectin 18 g l -1 0.05 % other 

3 and 7 days after treatments, but 9 days after 1st treatment it was found statistical 

differences between them (Table 3). 

The number of two-spotted spider mites was significant less among Abamectin 

18 g l -1 0.12 % and standards Azadirachtin A 10 g l -1 0.5 % 3 days after 1st treatment 

and Abamectin 18 g l -1 0.12 % and Azadirachtin A 10 g l -1 and Spirodiclofen 240 g l -1 

0.05 % 9 days after 1 st treatment. 

Table 3. Abundance of two-spotted spider mite (Tetranychus urticae) on cucumbers 

after first application 

3 lentelė. Paprastųjų voratinklinių erkių (Tetranychus urticae) gausumas agurkuose po 

pirmojo purškimo 

Treatments 

Variantas 

before 

treatment 

prieš 

purškimą 

Number of mites per leaf 

Erkių skaičius ant lapo 

3 days after 7 days after 

treatment treatment 

praėjus 3 dienoms praėjus 7 dienoms 

po purškimo po purškimo 

Babtai, 2005–2006 

9 days after 

treatment 

praėjus 9 dienoms 

po purškimo 

1 2 3 4 5 

2005 

Untreated 

40.25 58.0 c 62.5 b 59.25 e 

Nepurkšta 

Abamectin 0.12 % 31.75 4.5 a 1.0 a 0 a 

Abamectin 0.10 % 34.25 10.0 ab 1.75 a 0.5 abc 

Abamectin 0.075 % 33.75 11.0 ab 3.25 a 0 ab 

Abamectin 0.05 % 28.50 12.75 ab 2.75 a 10.5 d 

Azadirachtin A 0.5 % 27.0 21.0 b 6.50 a 10.0 cd 

Spirodiclofen 0.05 % 31.50 23.5 b 5.75 a 9.25 bcd 

49



1 2 3 4 5 

2006 

Untreated 

34.25 38.0 d 64.50 c 89.50 c 

Nepurkšta 

Abamectin 0.12 % 37.00 2.50 a 0.75 a 1.25 a 

Abamectin 0.10 % 36.50 4.0 ab 2.25 ab 2.0 a 

Abamectin 0.075 % 33.75 8.25 abc 3.0 ab 5.0 ab 

Abamectin 0.05 % 37.50 11.50 abc 4.25 ab 6.0 ab 

Azadirachtin 0.5 % 38.0 17.75 c 12.25 b 8.75 b 

Spirodiclofen 0.05 % 35.50 15.0 bc 7.50 ab 8.25 b 

Note: Means followed by the same letter are not different significantly (P = 0.05) according 

to Duncan’s multiple range test 


esmės nesiskiria 

The abundance of two-spotted spider mites in 2006 grew not so fast as in 2005. 

There were not found any statistical differences of the number of two-spotted spider 

mites between Abamectin 18 g l -1 0.12 % and Abamectin 18 g l -1 (0.10, 0.075, 0.05%) 

3, 5, 7 and 14 days after treatments. The number of two-spotted spider mites was 

significant less among Abamectin 18 g l -1 (0.12, 0.1, 0.075, 0.05 %) and standards 

Azadirachtin A 10 g l -1 0.5 % and Spirodiclofen 240 g l -1 0.05 % 3 days after 1 st treatment. 

There were found statistical differences of number among Abamectin 18 g l -1 

0.12 % and standards Azadirachtin A 10 g l -1 0.5 % and Spirodiclofen 240 g l -1 0.05 % 

3, 7 and 14 days after 1st treatment and Abamectin 0.1 % and Azadirachtin A 10 g l -1 

14 days after 1st treatment. 

Abamectin 18 g l -1 0.12 % was very efficient against two-spotted spider mite: 99.1, 

98.4 and 100 % 5, 7 and 9 days after 1st treatment in 2005 (Table 4). An efficiency of 

Abamectin 18 g l -1 (0.1 and 0.075 %) after 1st treatment against two-spotted spider 

mite was over 90 %. Effect of Abamectin 18 g l -1 0.05 % against mites was lower. 

Abamectin 18 g l -1 0.12 % was very efficient against two-spotted spider mite: 93.9, 

98.9, 98.7 and 98.1 % – 3, 7, 14 and 21 days after 1st treatment in 2006. Efficiency 

of Abamectin 18 g l -1 (0.1 and 0.075 %) after 1st treatment against two-spotted spider 

mite was over 90 %. 

There were found statistical differences of number of mines in leaf between all doses 

of Abamectin 1.8 g l -1 and untreated plants 7, 9 and 10 days after treatment (Table 5). 

The number of mines in leaf was significant less in Abamectin 1.8 g l -1 (0.12 and 

0.1 %) treated plants compare with Azadirachtin A 50 g l -1 and Spirodiclofen 240 g l -1 

treated plants 7, 9 days after treatment in 2005. The number of mines was significant 

less in Abamectin 0.12 % treated plants than Azadirachtin A and Spirodiclofen treated 

plants after 7 and 14 days after treatment in 2006. There were not found any statistical 

differences of number of mines in leaf between Abamectin all doses, except the highest 

and lowest doses after 9 days after treatment in 2005. 

50

Table 4. Efficiency of insecticide Abamectin applied against two-spotted spider 

mite (Tetranychus urticae) after first application 

4 lentelė. Insekticido abamektino efektyvumas nuo paprastųjų voratinklinių erkių 

(Tetranychus urticae) po pirmojo purškimo 

Treatments 

Variantas 

3 days after treatment 

praėjus 3 d. po purškimo 

Biological efficiency 




Babtai, 2005–2006 



2005 

Untreated 

- - - 

Nepurkšta 

Abamectin 0.12 % 92.24 98.40 100 

Abamectin 0.10 % 82.76 97.20 99.16 

Abamectin 0.075 % 81.03 94.80 100 

Abamectin 0.05 % 78.02 95.60 82.28 

Azadirachtin 0.5 % 63.79 89.60 83.12 

Spirodiclofen 0.05 % 59.48 90.80 84.39 

2006 

Untreated 

- - - 

Nepurkšta 

Abamectin 0.12 % 93.91 98.93 98.71 

Abamectin 0.10 % 90.12 96.73 87.40 

Abamectin 0.075 % 77.97 95.28 81.45 

Abamectin 0.05 % 72.36 93.98 72.40 

Azadirachtin 0.5 % 57.90 82.88 61.49 

Spirodiclofen 0.05 % 61.92 88.78 60.59 

Table 5. Damaged leaves of miner fly larvae on cucumbers after the first application 

5 lentelė. Minamusių lervų pažeisti lapai po pirmojo purškimo 

Treatments 

Variantas 

before 7 days after 

treatment treatment 

prieš praėjus 7 d. 

purškimą po purškimo 

Number of mines in leaf 

Minų skaičius lape 

2005 2006 

9 days after 

treatment 

praėjus 9 d. 

po purškimo 

before 

treatment 

prieš 

purškimą 

Babtai, 2005–2006 

7 days after 

treatment 

praėjus 7 d. 

po purškimo 

14 days after 

treatmen 

praėjus 14 d. 

po purškimo 

1 2 3 4 5 6 7 

Untreated 

0.50 0.42 d 0.45 f 0.35 0.27 d 0.30 d 

Nepurkšta 

Abamectin 0.12 % 0.55 0.12 a 0.12 a 0.32 0.05 a 0.10 a 

51

Abamectin 0.10 % 0.42 0.12 a 0.15 ab 0.37 0.07 ab 0.12 ab 

Abamectin 0.075 % 0.45 0.20 ab 0.22 abc 0.27 0.10 ab 0.15 abcd 

Abamectin 0.05 % 0.52 0.22 ab 0.25 bcd 0.25 0.12 ab 0.15 abcd 

Azadirachtin A 0.5 % 0.45 0.32 bcd 0.37 def 0.30 0.20 bcd 0.25 bcd 

Spirodiclofen 0.05 % 0.45 0.30 bcd 0.35 cdef 0.32 0.20 bcd 0.22 abcd 

Note: Means followed by the same letter are not different significantly (P = 0.05) according 

to Duncan’s multiple range test 


esmės nesiskiria. 

Abamectin 18 g l -1 0.12 % effectively reduced the number of mines in leaves 

(Table 6). Efficiency of Abamectin 18 g l -1 (0.1, 0.075 and 0.05 %) was lower. Efficiency 

of Azadirachtin A 50 g l -1 and Spirodiclofen 240 g l -1 was low. The lowest efficiency 

of almost all treated active ingredients was 14 days after treatment. 

Table 6. Efficiency of insecticide Abamectin against miner flies after the first 

application 

6 lentelė. Insekticido abamektino efektyvumas nuo minamusių po pirmojo purškimo 

Treatments 

Variantai 

7 days after 

treatment 

praėjus 

7 dienoms 

po purškimo 



2005 2006 

52 

9 days after 

treatment 

praėjus 

9 dienoms 

po purškimo 

7 days after 

treatment 

praėjus 

7 dienoms 

po purškimo 

Babtai, 2005–2006 

14 days after 

treatment 

praėjus 

14 dienų 

po purškimo 

Untreated 

- - - - 

Nepurkšta 

Abamectin 0.12 % 71.43 75.80 80.30 63.60 

Abamectin 0.10 % 65.99 60.40 75.26 62.20 

Abamectin 0.075 % 47.09 54.40 51.39 35.90 

Abamectin 0.05 % 46.63 46.60 48.15 30.70 

Azadirachtin 0.5 % 15.35 8.50 13.53 3.40 

Spirodiclofen 240 SC 0.05 % 28.58 13.40 18.80 19.80 

It was no observed negative direct effect on crop, beneficial fauna or pest resistance 

of insecticide Abamectin 18 g l -1 . 

Discussion. The number of various acaricides and insecticides were tested against 

two-spotted spider mite, Tetranychus urticae Koch. in different countries (Herron 

and Rophail, 1998; Nauen et al., 2001; Castagnoli et al., 2005). Three active ingredients 

from different chemical classification were tested in this study. The higher rates 

of Abamectin 1.8 g l -1 (0.12, 0.1, 0.075 %) effectively protected cucumbers against 

mites 14 days after treatment. Lowest rate (0.05 %) of Abamectin less affected mites, 

but statistically this effect differed only 9 days after first application in 2005. Only 

the highest rates of Abamectin (0.12, 0.1 %) efficiently reduced damage of miner fly

larvae. Azadirachtin A 10 g l -1 and Spirodiclofen 240 g l -1 were less effective against 

two-spotted spider mite and not enough effective against miner flies. Spirodiclofen 

240 g l -1 (0.5 l/ha) was less effective too, when in strawberry this insecticide (0.4 l/ha) 

was high efficient against two-spotted spider mite (Raudonis et al., 2005). 

According to Ochoa and Carballo (1993) Abamectin applied at the commercially 

recommended dosage, showed a satisfactory level of control of the eggs and larvae of 

Liriomyza huidobrensis. Lower dosages were less effective, like in our study. Abamectin 

provides residual activity in the field because of its translaminar action and rapid penetration 

of leaf tissue (Dybas, 1989), maybe this proposition explain why this active 

ingredient is more effective against leaf miner flies than Azadirachtin and Spirodiclofen. 

One of the new methods according to Luksienė et al. (2005) results in exposition of 

insects to the bait, containing hematoporphyrin dimethyl ether and following irradiation 

with visible light resulted in total killing of Liriomyza bryoniae adults. 

The ability of T. urticae to develop resistance to several acaricides has caused 

problems in many countries involved in agricultural production during the past 40 years 

(Herron, Rophail, 1993; Hinomoto, Takafuji, 1995; Stumpf, Nauen, 2001). Because of 

resistance problems it is more difficult to control leaf miner flies worldwide (Parrella 

et al., 1985). After 7–8 years of widespread commercial use of abamectin in pear 

orchards all T. urticae populations collected from pear showed low to moderate levels 

of resistance (Beers et al., 1998). The percentage of resistant mites to Abamectin 

decreased to levels equal or lower than 15 % in six months (Sato et al., 2005). Among 

T. urticae populations no resistance against abamectin was found in a population than 

had been subjected to fewer than 6 applications per year (Campos et al., 1995). In our 

experiment there was no observed negative direct effect on crop, beneficial fauna or 

pest resistance of insecticide Abamectin 18 g l -1 . Trumble and Morse (1993) reported 

that the best economic returns were generated by abamectin against T. urticae in 

combination with Phytoseiulus persimilis. This indicates that the chemical application 

of two-spotted spider mite can be successfully integrated with biological control. 

Abamectin is not only highly effective but it comes from biological source, which can 

be used in an environmentally friendly manner (have relatively low toxicity to many 

non-target arthropods) in integrated pest management programs (Dybas, 1989). 

Conclusions. 1. Efficiency of insecticide Abamectin 18 g l -1 0.12 % against twospotted 

spider mite was 92.24–100 %, against leaf miner flies it was 63.6–80.3 %. 

Abamectin 18 g l -1 0.12 % was effective against two-spotted spider mite till 14 days, 

against leaf miner flies – 9 days. 

2. Efficiency of insecticide Abamectin 18 g l -1 0.1 % against two-spotted spider 

mite was 82.8–99.2 %, against leaf miner flies – 60.4–75.26 %. Abamectin 18 g l -1 

0.10 % was effective against two-spotted spider mite till 14 days, against leaf miner 

flies – 7 days. 

3. Efficiency of insecticide Abamectin 18 g l-1 0.075 % against two-spotted spider 

mite was 81.0–100 %, against leaf miner flies – 35.9–54.4 %. Abamectin 18 g l -1 


flies was not enough effective. 

53

4. Efficiency of insecticide Abamectin 18 g l -1 0.05 % against two-spotted spider 

mite was 72.4–94.0 %, against leaf miner flies – 30.7–49.6 %. Abamectin 18 g l -1 


flies was not enough effective. 

Gauta 2009 06 30 


References 

1. Anon. EPPO Standards. 1997. Guidelines for the efficacy evaluation of plant protection 

products. Insecticides & Acaricides. Editor European and Mediterranean 

Plant Protection Organization, Paris. 

2. Beers E. H., Riedl H., Dunley J. E. 1998. Resistance to Abamectin and reversion 

to susceptibility to Fenbutatinoxide in spider-mite (Acari: Tetranychidae) 

populations in the Pacific-Northwest. Journal of Economic Entomology, 91: 

352–360. 

3. Boom C. E. M., Beek T. A., Dicke M. 2003. Differences among plant species 

in acceptance by the spider mite Tetranychus urticae Koch. Journal of Applied 

Entomology, 127(3): 177–183. 

4. Campos F., Krupa D. A., Dybas R. A. 1995. Susceptibility of population of 

two-spotted spider mite (Acari: Tetranychidae) populations in California to 

Abamectin. Journal of Economic Entomology, 88(2): 225–231. 

5. Castagnoli M., Liguori M., Simoni S., Duso C. 2005.Toxicity of some insecticides 

to Tetranychus urticae, Neoseiulus californicus and Tydeus californicus. 

Biocontrol, 50: 611–622. 

6. Dybas R. A. 1989. Abamectin use in crop protection. In Champbell W. C. (ed.) 

Ivermectin and abamectin. Springer, NY, USA. 

7. Fry J. D. 2000. Evolutionary adaptation to host plants in a laboratory population 

of the phytophagous mite Tetranychus urticae Koch. Oecologia, 81: 559–565. 

8. Helle W., Sabelis M. W. 1985. Spider Mites. Their Biology, natural Enemies and 

Control. Elsevier, Amsterdam, Oxford, New York, Tokyo, 1 A: 391–395. 

9. Herron G. A., Rophail J. 1998. Tebufenpyrad (Pyranica®) resistance detected 

in two-spotted spider mite Tetranychus urticae Koch. (Acarina: Tetranychidae) 

from apples in Western Australia, Experimental and Applied Acarology, 22: 

633–641. 

10. Hinomoto N., Takafuji A. 1995. Genetic changes in the population structure of 

the two-spotted spider mite, Tetranychus urticae Koch. (Acari: Tetranychidae) 

on vinyl-house strawberries. Applied Entomology and Zoology, 30: 521–528. 

11. Z. Luksienė, V. Buda, S. Radziutė. 2005. Effects of visible-light-activated hematoporphyrin 

dimethyl ether on the survival of leafminer Liriomyza bryoniae. 

Ekologija, 3: 17–21. 

54

12. Meier U. 1997. Growth stages of Mono- and Dicotyledonous plants. BBCH 

Monograph. Berlin: Blackwell Wissenschafts-Verlag. 

13. Nachman G., Zemek R. 2002. Interactions in a tritrophic acarine predator-prey 

metapopulation system III: Effects of Tetranychus urticae (Acari: Tetranychidae) 

on host plant condition. Experimental and Applied Acarology, 25: 27–42. 

14. Nauen R., Stumpf N., Elbert A., Zebitz C. P. W., Kraus W. 2001. Acaricide toxicity 

and resistance in larvae of different strains of Tetranychus urticae and Panonychus 

ulmi (Acari: Tetranychidae). Pest Management Science, 57: 253–261. 

15. Ochoa P., Carrballo M. 1993. Effect of various insecticides on Liriomyza huidobrencis 

(Diptera: Agromyzidae) and its parasitoid Diglyphus isaea Walker 

(Hymenoptera: Eulophidae). Manejo Integrado de Plagas (Costa Rica), 26: 

8–12. 

16. Ostraukas H., Pakalniškis S., Taluntytė L. 2005. Dipte-rous miners collected in 

greenhouse areas in Lithuania. Ekologija, 2:22-29. 

17. Ostraukas H., Pakalniškis S., Taluntytė L. 2003. The species composition of plant 

miningdipterous (Insecta: Diptera) of greenhouse surroundings in Lithuania. 

Ekologija, 3: 3–11. 

18. Pakalniškis S., Ostrauskas H., Taluntytė L. 2005. Dvisparniai vabzdžiai šiltnamio 

ir daržo kultūrinių augalų minuotojai. Vilnius. 

19. Park Y.-L., Lee J.-H. 2005. Impact of two-spotted spider mite (Acari: 

Tetranychidae) on growth and productivity of glasshouse cucumbers. Journal of 

Economic Entomology. 98: 457–463. 

20. Parrella M. P., Jones V. P., Youngman R. R., Lebeck L. M. 1985. Effect of leaf 

mining and leaf stippling of Liriomyza spp. on photosynthetic rates of chrysanthemum. 

Annals Entomological Society of America, 78: 90–93. 

21. Raudonis L., Valiuškaitė A., Survilienė E. 2005. Effects of Spirodiclofen on 

the Two-spotted Spider Mite, Tetranychus urticae (Acari: Tetranychidae) in 

Strawberries. Sodininkystė ir daržininkystė, 24(2): 43–53. 

22. Sato M. E., Silva M. Z., Raga A., Souza Filho M. F. 2005. Abamectin resistance 

in Tetranychus urticae Koch. (Acari: Tetranychidae): selection, cross-resistance 

and stability of resistance. Neotropical Entomology, 34(6): 991–998. 

23. Stumpf N., Nauen R. 2001. Cross resistance, inheritance, and biochemistry 

of mitochondrial electron transport inhibitor-acaricide resistance in Tetranychus 

urticae (Acari: Tetranychidae). Journal of Economic Entomology, 94: 

1 577–1 583. 

24. Trumble J. T., Morse J. P. 1993. Economics of integrating the predaceous mite 

Phytoseiulus persimilis (Acari: Phytoseiidae) with pesticides in strawberries. 

Horticultural Entomology, 86: 879–885. 

25. Van de Vrie M., McMurtry J. A., Huffaker C. B. 1972. Biology, ecology and pest 

status, and host-plant relationships of Tetranychids. Hilgardia, 41: 343–432. 

55


Abamektino poveikis paprastosioms voratinklinėms erkėms ir 

minamusėms šiltnamio agurkuose 

L. Duchovskienė 

Santrauka 

2005–2006 m. tirtas abamektino 18 g l -1 poveikis paprastosioms voratinklinėms erkėms 

(Tetranychus urticae Koch.) ir minamusėms (Liriomyza strigata, Liriomyza bryoniae) šiltnamio 

agurkuose. Abamektino 18 g l -1 0,12 % ir 0,1 % koncentracijų efektyvumas nuo paprastųjų 

voratinklinių erkių buvo atitinkamai 92,24–100 % ir 82,8–99,2 %. Abamektino 18 g l -1 0,075 % 

ir 0,05 % koncentracijų efektyvumas nuo paprastųjų voratinklinių erkių buvo atitinkamai 

81,0–100 % ir 72,4–94,0 %. Abamektino 18 g l -1 0,12 % ir 0,1 % koncentracijų efektyvumas 

nuo minamusių buvo atitinkamai 63,6–80,3 % ir 60,4–75,26 %. Abamektino 18 g l -1 0,075 % ir 

0,05 % koncentracijų efektyvumas nuo minamusių buvo atitinkamai 35,9–54,4 % ir 30,7–49,6 

%. Azadirachtino A 10 g l -1 0,5 % koncentracijos ir spirodiklofeno 240 g l -1 0,05 % koncentracijos 

efektyvumas nuo paprastųjų voratinklinių erkių ir minamusių buvo mažesnis. 

Reikšminiai žodžiai: abamektinas, agurkai, azidirachtinas A, Liriomyza bryoniae, 

Liriomyza strigata, spirodiklofenas, Tetranychus urticae. 

56




Different fungicide combinations against apple scab 

helping to avoid fungus resistance 

Maija Eihe, Regina Rancane, Liga Vilka 

Latvian Plant Protection Research Center, Lielvardes iela 36/38, Riga, LV 1006, 

Latvia, e-mail regina.rancane@laapc.lv 

IOBC guidelines for integrated fruit production prescribe the use of forecasting systems 

in direct plant protection. In Latvia, LPPRC, model RIMpro for apple scab (Venturia inaequalis) 

control was tested from 2003. Following to FRAC guidelines, in order to reduce the 

risk of fungus resistance developing, from 2007 efficacy of fungicide mixtures (Chorus, a. 

i. cyprodinil + Dithane NT, a. i. mancoceb; Effector, a. i. dithianon + Candit, a. i krezoxymmethyl) 

and alternately curative or strobilurine – protective fungicide use was tested. In all 

cases the first protective application before scab ascospores discharge was carried out with 

Cu product Champion 50. In case of emergency Effector was used during secondary scab 

infection period. 

Fungicides registered in Latvia for apple scab control were effective used as mixture 

of protective/curative or strobilurine products, no alternately, except strobilurine Candit (Qo 

inhibitor), which was applied separately, because fungus resistance appeared in the 3rd season 

of Candit use. Efficacy of Candit/Effector mixture corresponded other treatments. Curative 

product Chorus did not lost its efficacy after 6 seasons of use, applied no more than 3 times 

per season. Nevertheless, further strategy of resistance preclusion has to be considered, what 

request minimal use of risky products separately. In all cases fungicide applications, even of 

Chorus/Dithane mixture, were more effective before infection. Weather forecast not always 

was precise; in such cases the number of necessary applications increased. Most frequently 

under Latvia conditions there are three or four severe scab infection periods during the total 

primary infection period, subsequently 3 or 4 fungicide applications were necessary in addition 

to the first Champion treatment. 

Key words: apple scab, fungicides, mixtures, resistance, risk groups, use in alternation. 

Introduction. Development of pathogenic fungi resistance to several fungicides 

has become a considerable problem in plant protection, including apple scab (Venturia 

inaequalis (Cooke) Wint.) control in apple orchards. Fungicide Resistance Action 

Committee (FRAC) has developed fungicide resistance management guidelines to 

prolong the effectiveness of risky fungicides. The problem relates mainly to site-specific 

products disrupting single metabolic processes of definite fungus. In control of 

apple scab the most harmful products are demethylation inhibitors in sterol biosynt- 

57

hesis (SBI, DMI) (Scheinpflug, 1988; Szkolnik, 1981), strobilurins – quinone outside 

inhibitors (QoI) (Broniarek-Niemec, Bielenin, 2008; Turechek, Köller, 2004), 

and anilinopirimidines (Dux et al., 2004; Köller, Wilcox et al., 2005). According 

to anti-resistance management strategies, risky products are used in mixtures or in 

alternation with protective low risk fungicides, e. g. mancozeb, also they are used in 

sufficiently high dosages, only when justified and mainly protectively (Köller, Parker 

et al., 2005; Köller, Wilcox, 2001, Staub, 1991). FRAC is developed fungicide Code 

List according to resistance risk level (FRAC Code List, 2007). 

In Latvia traditionally minimal amount of pesticides was used on horticultural 

crops. During the last decades, apple plantations quickly enlarged and growers intensified 

pesticide use to save yield quality. The choice of fungicides is poor. Therefore, 

recommendations of suitable anti-resistance plant protection strategy are important. 

From 2003 apple scab warning model RIMpro was tested in Latvian Plant 

Protection Research Centre for adoption under conditions of Latvia. In the first years 

only curative fungicide Chorus 75 WG (a. i. cyprodinil, anilino pyrimidine group) 

was used shortly after RIMpro warning about risky infection. During the last years, 

FRAC (Fungicide Resistance Action Committee) baselines and recommendations about 

strategies to prevent fungus resistance were considered: use of fungicide mixtures with 

different modes of action or their use in alternation, as much as possible not several 

times in turn, use of protective products before forecasted infection as well as all applications 

before infection. There was little experience of fungicide mixtures efficacy 

in apple scab control in Latvia up till now. In 2007 and 2008 field trial was arranged 

to compare efficacy of different fungicide combinations in scab control taking into 

account RIMpro indices of risky infection. 

Object, methods and conditions. Field trial was carried out in peasant farm 

“Kalnanoras”, Ikskile, Ogre district in 2007–2008. There was investigated apple cultivar 

‘Spartan’, on dwarf rootstock B 396, planted in 2000. Trial plot: 6 trees, 4 replicates 

in randomized blocks. Untreated control plots located out of blocks. In the trial 

the mostly used fungicides registered for apple orchards in Latvia and recommended 

mixtures were employed (Table 1). 

Table 1. Fungicides used in trial 

1 lentelė. Bandyme panaudoti fungicidai 

Trade name 

Prekinis 

pavadinimas 

Active 

ingredient 

Aktyvi sudedamoji 

dalis 

g kg -1 

Chemical 

group 

Cheminė 

grupė 

Mode of action 

Poveikio būdas 

FRAC Resistance 

code risk 

FRAC Atsparumo 

kodas pavojus 

1 2 3 4 5 6 7 

Champion 50 SP Copper hydroxide 770 Copper Protecting 

M1 Low 

Vario hidroksidas Varis Apsauginis 

Mažas 

Dithane NT WG Mancozeb 750 Dithiocarbamate 

Protecting 

Apsauginis 

M3 Low 

Mažas 

Effector WG Dithianone 700 Quinone Protecting 

Apsauginis 

M9 

Low 

Mažas 

58



1 2 3 4 5 6 7 

Chorus 75 WG Cyprodinil 750 Anilinopirimidine 

Systemic, post-infection 

Sisteminis, poinfekcinis 

9 Medium 

Vidutinis 

Candit WG Kresoxim-metyl 500 Strobilurin, 

QoI 

Locally systemic, 

mostly protecting 

Vietiškai sisteminis, 

daugiausia apsauginis 

11 High 

Didelis 

These are mostly all fungicides registered in Latvia for apple scab control with 

exception of Score 250 (a. i. difenoconazole, SBI, DMI), against which considerable 

fungus resistance was observed in trial orchard after 5 years of use. 

In anti-resistance management strategy the prophylaxis in advance of primary 

apple scab infection period at green tip stage of apple trees with protecting spray is 

recommended (Köller, Parker et al., 2005). In the trial this application was carried out 

with copper fungicide Champion 50 in all treatments. 

All products were used in maximal rates registered in Latvia: Champion 

4.0 (a. i. 3.08) kg ha -1 , Dithane 3.0 (a. i. 2.25), Effector 0.6 (a. i. 0.42), Chorus 

0.3 (a. i. 0.225), Candit 0.25 (a. i. 0.125) kg ha -1 . In mixture 75 % of each product was 

used. Water volume was 600 l ha -1 . The biggest attention was paid to primary scab 

infection control. If effectiveness of applications was insufficient, fungicide use was 

continued during secondary scab infection period. 

Real beginning, intensity and the end of ascospores release was determined on 

spore traps – glass slides placed on old apple leaf layer in trial orchard. Slides were 

collected after every rain and spores counted under microscope. Discharge of scab 

ascospores commonly is ending in the middle of June. 

In 2007 Champion was sprayed on 18.04. (GS 53, green tip) in all the treatments, 

except untreated control. The first scab ascospores were released on 20.04. 

In 2007 the trial was carried out using systemic fungicide Chorus (alone, no 

more than 3 times per primary infection period), mixture of Chorus and protective 

Dithane 1–2 days after RIMpro showed risky infection level (above 70 RIM), as well 

as Dithane and Chorus in alternation accordingly shortly before and after infection. 

Before or shortly after infection (24 hours) protective Efector, mixture of Efector and 

strobilurine Candit, as well as Effector and Candit (2 times) in alternation were used 

(Table 2). 

RIMpro shown the amount of efficient residues (more than 25 %) after each 

application was taken in account too. Chorus in trial territory up till now was used for 

6 years 2–3 times per season without symptoms of efficacy decrease, Candit – for 3 

years with efficacy decrease symptoms. 

59

Table 2. Fungicide combinations for control of primary apple scab infection, 

2007 

2 lentelė. Fungicidų deriniai pirminės obelų rauplių infekcijos kontrolei, 2007 m. 

Dates of critical infection 

periods 

Kritinių infekcijos 

laikotarpių datos 

RIM risk values per day in 

average 

Vidutinės RIM rizikos vertės per 

dieną 

Precipitation sum 

Kritulių suma (mm) 

Dates of fungicide applications 

Fungicido panaudojimo datos 

Apple growth stages (GS), 

BBCH 

Obelų augimo tarpsniai (AT) 

Treatments 

Variantai 

Control – untreated 

Kontrolė – neapdorota 

Chorus (Ch) after infection 

Chorus (Ch) po infekcijos 

Dithane (D) before, Chorus after 

infection 

Dithane (D) prieš, Chorus po 

infekcijos 

Dithane / Chorus mixture after 

infection 

Dithane / Chorus mišinys po infekcijos 

Effector (E) before forecasted or 

shortly after infection 

Effector (E) prieš numatytą infekciją 

arba tuoj po jos 

Effector / Candit mixture for 

3 times 

Effector / Candit mišinys 3 kartus 

Effector in turn with Candit (Ca) 

Effector pakaitomis su Candit (Ca) 

09–10.05 15.05 26–28.05 01.06 

832 337 652 117 

17.0 13.2 27.4 1.4 

27.04 10.05 16.05 24.05 28.05 30.05 

55 

(mouseear) 

(žiediniai 

pumpurai 

dar 

uždari) 

57 

(red 

buds) 

(raudoni 

pumpurai) 

59 

(ballon 

stage) 

(dauguma 

žiedų su 

vainiklapiais) 

65 

(full 

flowering) 

(visiškas 

žydėjimas) 

67 

(end of 

flowering) 

(žiedai 

vysta) 

69 

(end of 

flowering) 

(žydėjimo 

pabaiga) 

- - - - - - 

- Ch Ch - Ch - 

D Ch Ch - Ch - 

- D/Ch D/Ch - D/Ch - 

E E - E - E 

E/Ca E/Ca - E/Ca - E 

E Ca - Ca - E 

Weather conditions during May were rainy, especially in the 3rd 10-day period, 

when precipitation sum was 49.8 mm, 293 % from normal, and mean temperature 

60

19.4° C, 6.7 °C above normal, being the warmest period in Latvia during 84 years. 

Such conditions promoted primary scab infection. Rainy period started again from 

the middle of June and July and additional fungicide application during secondary 

infection period was necessary. Additional applications with Effector were carried out 

on 19.06 and 12.07 in the 7 th treatment where Candit alone 2 times was used, and on 

27.07 in all treatments. It is permitted to use the low resistance risk product Effector 

for 6 times per season in Latvia. 

In 2008 all fungicide combination schedules were repeated, except 7 th treatment, 

where clearly appeared efficacy loss; strobilurine Candit was used alone for 2 times. 

Following to FRAC recommendations, all fungicide treatments were carried out before 

forecasted precipitation and scab infection (Table 3). 

Table 3. Fungicide combinations for control of primary apple scab infection, 

2008 

3 lentelė. Fungicidų deriniai pirminės obelų rauplių infekcijos kontrolei, 2008 m. 

Dates of precipitation 

periods 

Kritulių laikotarpių datos 

Precipitation sum 

Kritulių suma (mm) 

Dates of fungicide applications 

Fungicido panaudojimo 

datos 

Apple growth stages 

(GS), BBCH 

Obelų augimo tarpsniai 

(AT) 

02–05.05 08, 12.05 15–18.05 11–20.06 

2.2 0.4 14.2 26.8 

29.04 09.05 13.05 10.06 

56 

(green buds / 

žali 

pumpurai) 

58 

(pink buds / 

rausvi 

pumpurai) 

61 

61 

(beginning of 

flowering / 

žydėjimo pradžia) 

71 

(fruits of 10 mm in 

diameter / 

10 mm skersmens 

vaisiai) 

Treatments 

Variantai 

Control – untreated 

- - - - 

Kontrolė – neapdorota 

Chorus (Ch) Ch - Ch Ch 

Dithane (D) in turn with D Ch - D 

Chorus 

Dithane (D) kartu su 

Chorus 

Dithane / Chorus mixture D/Ch D/Ch - D/Ch 

Dithane / Chorus mišinys 

Effector (E) E E - E 

Effector/ Candit mixture E/Ca E/Ca - E/Ca 

Effector/ Candit mišinys 

In 2008 Champion was sprayed on 16.04. (GS 53) in all treatments including 

untreated control for partly decrease of high scab infection level in control plots. The 

first scab ascospores were released on 17.04.

Weather conditions in May were comparably dry and cool; some days it rained, 

noteworthy that on 18.05 it rained 12.2 mm and the mean temperature in the 2 nd 10-day 

period was 10 °C. The 3 rd 10-day period of May and the 1 st of June were completely 

without precipitation. Rainy period lasted from June 11 th until the middle of July. 

Apple scab widely spread in unprotected areas. In treated plots no fungicide applications 

were carried out during secondary infection period. For weather forecast data 

of Latvian Environment, Geology & Meteorology Agency (LEGMA) and web page 

www.gismeteo.ru was employed. 

Apple growth stages are characterized according to BBCH scale. 

Scab incidence and extent level was assessed on 100 leaves and fruits per plot 

from the beginning of disease appearance in control treatment, before each fungicide 

application and further after rainy periods, on leaves – till the end of July, on fruits – until 

harvesting. Disease extent was evaluated according to % scale of damaged surface. 

The data were subjected to analysis of variance. Significant differences at the 

probability of 95 % are shown in the tables by letters. Data of control treatment are 

not included in processing. 

Results. In 2007 there were four noteworthy critical primary apple scab infection 

periods. At the end of April – beginning of May precipitation appeared later than 

forecasted, subsequently the first test treatment was carried out too early in schedules 

with planned fungicide application shortly before infection (see Table 2). As a result, 

according to the schedule of planned applications, before infection four treatments 

were carried out and after infection – three fungicide treatments during primary apple 

scab infection period. 

Five tested schedules significantly decreased scab infection level on the leaves 

without significant difference among them, but in the 7 th treatment, where 2 times 

strobilurine Candit was used, there was clearly visible the loss of effectiveness, what 

indicated fungus resistance development. After two additional Effector sprays on 

19.06 and 12.07, scab infection leveled with other treatments until the end of July 

(Table 4). 

Table 4. Apple scab incidence on apple leaves using different fungicide combinations, 

Latvia, 2007 

4 lentelė. Obelų rauplių paplitimas ant obelų lapų naudojant skirtingus fungicidų derinius, 

Latvija, 2007 m. 

Dates of assessments 

Apskaitų datos 

Treatments 

24.05 19.06 06.07 27.07 

Apdorojimai 

scab incidence (amount of infected leaves) 

rauplių paplitimas (pažeistų lapų kiekis) (%) 

1 2 3 4 5 

Untreated control 

0.2 25.3 38.7 55.2 

Neapdorota kontrolė 

All schedules, except treatment with Candit 

Visos schemos, išskyrus apdorojimą su Candit 

0 0.60 a 0.55 a 3.35 a 

62



1 2 3 4 5 

Candit for 2 times separately in alternation 0 2.75 b 5.00 b 5.00 a 

with Effector 

Candit 2 kartus atskirai, kaitaliojant su Effector 

LSD 95 

/ R 95 

- 1.52 2.12 4.22 

According to fruit infection extension, the same tendency appeared as on the leaves: 

after the schedule of two sprays with Candit alone there was significantly higher 

scab infection level in the beginning of July in comparison with other schedules. There 

was also tendency of higher fruit infection in the treatment with Dithane/Chorus mixture 

for 3 times after primary infection periods, though on leaves, as infection source, 

such tendency did not appear. Possibly mixture with Dithane could be used before 

primary infection period. After additional fungicide applications during secondary 

scab infection period until the harvest time fruit infection leveled without significant 

difference between treatments. There remained the tendency of higher infection in 

Dithane/Chorus mixture treatment. The decrease of infection in Candit treatment was 

due to dropping of more damaged fruits (Table 5). 

Table 5. Apple scab incidence on apple fruits using different fungicide combinations, 

Latvia, 2007 

5 lentelė. Obelų rauplių paplitimas ant obuolių naudojant skirtingus fungicidų derinius, 


Treatments 

Variantai 


Apskaitų datos 

19.06 06.07 27.07 24.09 

scab incidence (amount of infected fruits) 

rauplių paplitimas (pažeistų vaisių kiekis) (%) 

29.0 35.0 59.9 82.3 



All schedules, except treatment with Candit 0 1.62 a 3.75 a 6.0 a 

use and Dithane/Chorus mixture 

Visos schemos, išskyrus apdorojimą su Candit 

ir Dithane/Chorus mišinį 

Dithane/Chorus mixture for 3 times 

0 2.5 ab 6.25 b 8.0 a 

Dithane/Chorus mišinys 3 kartus 

Candit for 2 times separately in alternation 0 5.0 b 5.25 ab 3.75 a 

with Effector 

Candit 2 kartus atskirai, kaitaliojant su Effector 

LSD 95 

/ R 95 

- 3.09 2.24 4.63 

In the tables there are shown only more characteristic results of 10 assessments. 

The extent of damaged leaf surface showed the same tendency as the amount of damaged 

fruits. 

63

In 2008 there were 4 noteworthy precipitation periods during primary scab infection 

period, like in 2007. Applying all fungicides 1–4 days before forecasted rain 

period effectiveness of all treatments was similar to scab infection control on the 

leaves in total. There was some significant difference between treatments in separate 

assessments, but the tendency did not maintain during assessment period. At the end 

of July there was no significant difference of leaf infection level between treatments 

(Table 6). Decrease of leaf infection level on 11.07 in comparison with previous assessment 

was due to forming young uninfected leaves. 

Table 6. Apple scab incidence on apple leaves using different fungicide combinations 

for 3 times before forecasted primary infection, 2008, Latvia 

6 lentelė. Obelų rauplių paplitimas ant obelų lapų naudojant skirtingus fungicidų derinius 

po 3 kartus prieš numatytą pirminę infekciją, Latvija, 2008 m. 

Treatments 

Apdorojimai 

Dates of assessmentsApskaitų datos 

10.06 04.07 11.07 31.07 

Scab incidence (amount of infected leaves) 

rauplių paplitimas (pažeistų lapų kiekis) (%) 

11.8 27.5 40.75 35.0 



Chorus 75 separately 

0 3.50 a 3.50 a 4.75 a 

Chorus 75 atskirai 

Dithane NT for 2 times, Chorus 1 time 0 3.25 a 2.75 ab 5.25 a 

Dithane NT 2 kartus, Chorus 1 kartą 

Mixture of Dithane/ Chorus 

0 4.25 ab 1.75 b 5.75 a 

Dithane/Chorus mišinys 

Effector separately 

0 5.50 b 2.50 ab 5.75 a 

Effector atskirai 

Mixture of Effector/ Candit 

0 4.00 ab 1.75 b 3.50 a 

Effector/Candit mišinys 

LSD 95 

/ R 95 

- 1.65 1.44 2.81 

For control of fruit infection the use of Chorus alone during primary scab infection 

period was significantly less effective. Such results did not indicate the appearance of 

fungus resistance because this schedule effectively controlled leaf infection, but postactivity 

of systemic product was too short to protect fruits until harvest (Table 7). 

64

Table 7. Apple scab incidence on apple fruits using different fungicide combinations, 

2008, Latvia 

7 lentelė. Obelų rauplių paplitimas ant obuolių naudojant skirtingus fungicidų derinius, 


Treatments 

Variantai 


Įvertinimo datos 

11.07 31.07 05.09 30.09 

scab incidence (amount of infected fruits) 

rauplių paplitimas (pažeistų vaisių kiekis) (%) 

12.5 21.0 63.5 76.8 



Chorus 75 separately 

0 0.75 a 5.25 a 7.50 a 

Chorus 75 atskirai 

Dithane/Chorus in alternation and 

0 0.25 b 4.50 a 3.88 b 

Mixture of Effector/Candit 

Dithane/Chorus pakaitomis ir Effector/Candit 

mišinys 

Mixture of Dithane/Chorus and Effector 0 0.12 b 2.75 b 4.38 b 

separately 

Dithane/Chorus mišinys ir Effector pakaitomis 

LSD 95 

/ R 95 

- 0.49 1.65 1.80 

Discussion. The use of protecting and risky fungicide mixtures during the primary 

scab infection period, according to the results of several authors, is the most effective 

component in apple scab protection schedules. Mixture of anilinopyrimidine with 

mancozeb has been more effective in comparison with mancozeb alone and especially 

anilinopyrimidine alone that provided unacceptable level of scab control (Köller, 

Wilcox et al., 2005; Köller, Parker et al., 2005). Such resolution agreed with our research 

results, by comparison of effectiveness of Dithane/ Chorus mixture and Chorus 

alone. According to strobilurins, several trial results showed that before resistance 

emergence strobilurin products alone in higher registered dosage were more effective 

in comparison with strobin/ mancozeb mixture and mancozeb alone (Turechek, Köller, 

2004). However, if the symptoms of resistance are observed, strobilurins will have 

to be used only in mixture with another scab fungicide (Köller, Parker et al., 2005). 

Protecting product dithianon (Effector WG registered in Latvia) is recommended for 

the mixture with krezoxym-metyl products by manufacturer company BASF. Therefore, 

this mixture was tested in our trial and although the decreased effect of separately 

used krezoxym-metyl (Candit WG) appeared already in the trial orchard the use of 

Effector/Candit in protection schedule had similar effectiveness in comparison with 

other schedules. 

Conclusions. 1. The use of protecting, low resistance risk fungicide Dithane 

NT (a. i. mancozeb) 1–4 days before primary apple scab infection in alternation with 

systemic, medium risky product Chorus 75 (a. i. cyprodinil) shortly before or after 

infection, as well as the use of Dithane/ Chorus mixture at dosage of 75 % from full 

rate of each product before infection in resistance preventive protection schedule 

similarly effectively controlled apple scab infection on leaves and fruits. 

65

2. High resistance risk strobilurin fungicide Candit (a. i. krezoxym-metyl) showed 

clearly visible loss of effectiveness used the 4th season for 2 times separately in 

alternation with Effector, while the use of Effector/Candit mixture for 3 times shortly 

before or after infection at dosage of 75 % from full rate of each product was similarly 

effective in comparison with other resistance preventive protection schedules. 

3. Medium risky product Chorus 75, used the 7 th and 8th season for 3 times by 

turns during primary scab infection period was similarly effective in resistance preventive 

schedules controling apple leaf infection, but lost the effect of fruit infection 

control during secondary scab infection period. Such schedule is not recommended 

in integrated plant protection. 

4. Low risk protecting fungicide Effector (a. i. dithianone) used for 3 times by 

turns shortly before or after primary scab infection was similarly effective with other 

resistance preventive schedules, but such schedule is not recommended in integrated 

plant protection. 

5. Planning fungicide treatments before infection can increase the number of 

sprays if weather forecast is not precise. 

Gauta 2009 06 30 


References 

1. Broniarek-Niemec A., Bielenin A. 2008. Resistance of Venturia inaequalis 

to strobilurin and dodine fungicides in Polish apple orchards. Zemdirbyste- 

Agriculture, 95(3): 366–372. 

2. Dux H., Sierotzki H., Gisi U. 2004. Sensitivity of Venturia inaequalis populations 

to anilinopyrimidyne, DMI and QoI fungicides. Abstracts14th International 

Reinhardsbrunn Symposium “Modern fungicides and antifungal compounds”, 

25–29 April, 2004, Germany, 40–41. 

3. FRAC Code List 2007: Fungicides sorted by mode of action (including FRAC 

Code numbering) (http://www. Frac.info/frac/publication) 

4. Köller W., Wilcox W. F. 2001. Evidence for predisposition of fungicide-resistant 

isolates of Venturia inaequalis to a preferential selection for resistance to other 

fungicides. Phytopathology, 91: 776–781. 

5. Köller W., Parker D., Turechek W., Rosenberger D., Wilcox W., Caroll J., 

Agnello A., Reissig H. 2005. Fungicide resistance of apple scab: status Quo and 

management options. New York Fruit Quarterly, 13(1): 9–17. 

6. Köller W., Wilcox W. F., Parker D. M. 2005. Sensitivity of Venturia inaequalis 

populations to anilinopyrimidine fungicides and their contribution to scab management 

in New York. Plant Disease, 89 (4): 357–365. 

7. Scheinpflug H. 1988. Resistance management strategies for using DMI fungicides. 

In: Fungicide resistance in North America C. J. Delp, ed. APS Press, St. 

Paul, MN, 93–94. 

66

8. Staub T. 1991. Fungicide resistance: Practical experience with antiresistance 

strategies and the role of integrated use. Annual Review of Phytopathology, 29: 

421–442. 

9. Szkolnik M. 1981. Physical modes of action of sterol-inhibiting fungicides 

against apple diseases. Plant Disease, 65: 981–985. 

10. Turechek W. W., Köller W. 2004. Managing resistance of Venturia inaequalis 

to the strobilurin fungicides (www.plantmanegementnetwork.org/pub/php/ 

research/2004/strobilurin). 


Įvairūs fungicidų deriniai nuo obelų rauplių, padedantys išvengti 

rauplėgrybio atsparumo 

M. Eihe, R. Rancane, L. Vilka 

Santrauka 

Auginant vaisius integruotai, jų produkcijai taikomos IOBC nuorodos rekomenduoja 

augalų apsaugai naudoti prognozavimo sistemas. Latvijoje, Augalų apsaugos tyrimų centre, 

nuo 2003 metų buvo tirtas RIMpro modelis obelų rauplių (Venturia inaequalis) kontrolei. 

Pagal FRAC nuorodas, siekiant sumažinti rauplėgrybio atsparumo išsivystymo pavojų, nuo 

2007 metų buvo tirtas fungicidų mišinių (Chorus, a. i. cyprodinil + Dithane NT, a. i. mancoceb; 

Effector, a. i. dithianon + Candit, a. i. krezoxym-methyl) ir pakaitomis naudojamų 

gydomojo arba strobilurino – apsauginio fungicido veiksmingumas. Visais atvejais pirmasis 

apsauginis apdorojimas prieš rauplių askosporų pasirodymą buvo atliktas su Cu produktu 

Champion 50. Esant reikalui, antrinės rauplių infekcijos laikotarpiu naudotas Effector. 

Latvijoje registruoti fungicidai nuo obelų rauplių buvo veiksmingi naudojami ne pakaitomis, 

o kaip apsauginių / gydomųjų arba strobilurino produktų mišinys, išskyrus strobiluriną 

Candit (Qo inhibitorių), kuris buvo naudojamas atskirai, nes trečiuoju Candit naudojimo sezonu 

atsidaro atsparumas rauplėgrybiams. Candit / Effector mišinio veiksmingumas buvo toks pat 

kaip ir kitų fungicidų. Gydomasis produktas Chorus po šešių naudojimo sezonų neprarado savo 

veiksmingumo, jei buvo panaudotas ne daugiau kaip 3 kartus per sezoną. Vis dėlto reikia apsvarstyti 

tolimesnę strategiją, kaip užkirsti kelią atsparumo išsivystymui, t. y. kaip minimaliai naudoti 

pavojingus produktus atskirai. Visais atvejais fungicidai, net Chorus/Dithane mišinys, buvo 

veiksmingesni prieš infekciją. Orų prognozės ne visada pasitvirtindavo – tada fungicidus tekdavo 

naudoti dažniau. Latvijos sąlygomis bendru pirminės infekcijos laikotarpiu būna trys ar keturios 

stiprios infekcijos, taigi, be pirmojo apdorojimo su Champion, reikėjo naudoti dar 3–4 fungicidus. 

Reikšminiai žodžiai: atsparumas, fungicidai, mišiniai, naudojimas pakaitomis, obelų 

rauplės, rizikos grupės, . 

67




Influence of growth regulators on seed germination 

energy and biometrical parameters of vegetables 

Julė Jankauskienė, Elena Survilienė 


Lithuania, e-mail j.jankauskiene@lsdi.lt 

Influence of growth regulators on seed germination energy and biometrical parameters 

of vegetables was investigated at the Lithuanian Institute of Horticulture in 2007. The seeds of 

cucumber ‘Krukiai’ F1, red beet ‘Joniai’, radish ‘Babtų žara’, tomato ‘Arvaisa’ F1 were soaked 

in the solutions of growth regulators Biojodis, Biokal 01, Bioforce, Agronom effect, Inzar, 

Oksichumat, Penergetic p. Control – the seeds soaked in water. After soaking seeds were sown 

into multicell flats, in which plants were grown for 30 days in greenhouse. It was established 

seed germination energy and seedling biometrical measurements (plant height, weight, leaf 

number, leaf area) were carried out. Plant growth regulators Oksichumat, Agronom effect, 

Bioforce, Penergetic p positively influenced seed germination energy of radish, tomato and 

the growth and development of cucumber, red beet, tomato and radish seedlings. 

Key words: biometrical parameters, cucumber, germination energy, growth regulator, 

radish, red beet, seedlings, seed soak, tomato. 

Introduction. Plant growth regulators are widely applied in the integrated plant 

growing for seed soaking. In vegetable growing, growth regulators also became 

more popular: for seed soaking, inflorescences spraying, shoot and plant watering 

or spraying through leaves. Growth regulators improve seed germination power, increase 

yield, plants become resistant to diseases and unfavourable growth conditions, 

produce yield earlyer, the yield becomes more qualitative (Halter et al., 2005; Kadiri 

et al., 1997; Papadopoulos et al., 2006; Saglam et al., 2002; Гущина, Къдрев, 1987). 

There are a lot of various growth regulators: Ivin, Natrium gumat, Uetinas, Ambiol, 

Biozin, Gibersib, Epin, Silk, ЭБФ-5, Oxydat, Penergetic p. They are of natural orign 

or synthetic. Kadiri et at. (1997) investigated the influence of natural growth regulators 

Indole-3-acetic, Gibberellic acid and Coconut milk on plant height, yield and the 

amount of vitamin C in fruits. It was established that applying these growth regulators 

plants were higher and the yield increased (Kadiri et al., 1997). According to the 

data of investigations, growth regulator Atonic not only increased the yield of sweet 

pepper, but also improved seed quality (Panajotov, 1997). 

Plant growth regulators are being used in greenhouses too. It was established that 

spraying tomatoes with Ruvit for three times during vegetation they started earlyer 

69

produce yield, the early and total yield increased (Гущина, Къдрев, 1987). In greenhouse 

spaying plants with Kinetin, the yield becomes more abundant and fruits – bigger 

(Papadopoulos et al., 2006). According to the data of the other investigators, the 

application of various biostimulators on cucumber in greenhouses positively influences 

plant growth (Boehme et al., 2005). There was investigated the influence of growth 

regulators mixed into fertilizator solution on plant productivity (Bugbee, White, 1984; 

Lopez-Elias et al., 2005). Moreover, there was investigated the influence of different 

growth regulators and their mixtures on plant productivity (Brocklehurst et al., 1982). 

Besides, growth regulators are being widely used for vegetable and flower seed soaking 

(Brigard et al., 2006; Magnitskiy et al., 2006; Pasian, Bennett, 1999). 

The aim of the study – to establish the influence of various natural growth regulators 

on vegetable seed germination power and shoot development. 


Institute of Horticulture, in greenhouse covered with double polymeric film in 

2007. The object of investigation – vegetable seeds: cucumber ‘Krukiai’ F 1 

, red beet 

‘Joniai’, radish ‘Babtų žara’, and tomato ‘Arvaisa’ F 1 

. For seed soaking these organic 

praparations were used: Biojodis, Biokal 01, Bioforce, Agronom effect, Inzar, Oksichumat, 

Penergetic p. Control – seeds soaked in water. Biojodis – preparation made 

on the basis of biohumus water extract, enriched with biologically active iodine, 

biotransformators and microelements. Biokal 01– fertilizer made out of medicinal 

herbs (57 %), biohumus extracts (38 %), mineral water, essential oils, macro- and microelements 

(5 %). Oksichumat – 10 % solution of huminic acids obtained out of peat 

or brown carbon. Penergetic-p – bioactivator, which activates plant cells participating 

in metabolism. Bioforce – extract out of Ascophyllum nodosum sea herbs. Agronom 

effect – plant growth stimulator, in the composition of which there is not less than 

700 g kg -1 of huminic acids salt, also silicon salt and microelements. Inzar – growth 

regulator, in the composition of which there is not less than 100 g 1 -1 of Maidenhair 

tree (Ginkgo biloba) extract. 

P r e p a r a t i o n o f s o l u t i o n s f o r s e e d s o a k i n g a n d d u r a t i o n 

o f s e e d s o a k i n g. 0.5 g of Agronom effect preparation was dissolved in 2 l of 

water and seeds were soaked in this solution for 6 h. 1 ml of Oksichumat was purred 

into 1 l of water and seeds were soaked in this solution for 10 h. Biokal 01 solution was 

prepared as follows: 3 parts of Biokal 01 solution + 10 parts of water. In this solution 

seeds were soaked for 48 h. 2 g of Penergetic p were solved in 1 liter of water and in 

this solution seeds were soaked for 10 h. In Biojodis solution seeds were soaked for 

10 h. 0.8 ml of Bioforce was purred into 1 l of water and seeds were soaked in this 

solution for 8 h. 1 ml of Inzar was purred into 0.5 l of water and seeds were soaked 

in this solution for 8 h. 

There was established germination energy of seeds soaked in the investigated 

solutions. Seeds were put into Petri plates (10 seeds per each) and moistened with 

the prepared solutions. After five days there were calculated germinated seeds. Seed 

germination energy was expressed in percents. After soaking the remained seeds were 

dried. Multicell flats (cup volume – 60 ml, in the flat – 64 cups) were filled with the 

prepared peat substratum (deacidified pH 5.5–6.5with fertilizers) and vegetable seeds 

soaked in various solutions were sown in them. Plants were grown in flats for 3 weeks. 

Then there was measured plant height, leaf assimilation area; there were calculated 

70

leaves and plant fresh weight established (weighting them). Leaf assimilation area was 

measured with leaf area measurer Win DIAS (Delta-T Devices, England). There were 

measured 10 plants per each variant. Experiment was carried out in three replications. 

Data processed by statistical methods (Tarakanovas, Raudonius, 2003). 

Results. Plant growth regulators didn’t influence positively germination energy 

of cucumber ‘Krukiai’ F 1 

and tomato ‘Arvaisa’ F 1 

seeds (Table 1). Nevertheless, 

germination energy of radish seeds soaked in plant growth regulator solutions was 

better than this of seeds soaked in water. Radish seeds soaked in solutions Bioforce, 

Biokal, and Penergetic p were distinguished for faster germination energy than seeds 

soaked in water. 

Table 1. Germination energy of vegetable seeds soaked in growth regulator solutions 

(%) 

1 lentelė. Daržovių sėklų, mirkytų įvairiuose augimo reguliatoriuose, dygimo energija, 

% 

Growth regulator 

Augimo reguliatorius 

Cucumber 

Agurkai 

‘Krukiai’ F 1 

Red beet 

Raudonieji burokėliai 

‘Joniai’ 

Radish 

Ridikėliai 

‘Babtų žara’ 

Tomato 

Pomidorai 

‘Arvaisa’ F 1 

Water 

100 90 70 100 

Vanduo 

Agronom effect 100 100 70 100 

Biojodis 100 100 100 100 

Bioforce 90 20 70 100 

Biokal 01 100 20 90 100 

Inzar 100 10 90 90 

Oksichumat 

90 70 80 40 

Oksigumatas 

Penergetic p 100 90 90 40 

Cucumber, which seeds were soaked in solutions Agronom effect, Biojodis, 

Bioforce, Inzar and Penergetic p, developed slightly faster than these, which seeds 

were soaked in water. They already had third real leaf, while cucumber, which seeds 

were soaked in water, had only two real leaves. Growth regulators Agronom effect, 

Bioforce, Inzar and Penergetic p mostly influenced cucumber height, fresh weight 

and leaf assimilation area (Table 2). Cucumber shoots, which seeds were soaked in 

solution Agronom effect, were 33 % higher, their fresh weight was 43.4 % bigger, and 

leaf assimilation area was two times bigger than this of schoots, which seeds were 


Red beet, which seeds were soaked in solutions Agronom effect, Bioforce, Inzar 

and Penergetic p, were correspondingly 20.9 %, 32.9 %, 21.9 % and 14.2 % higher 

than shoots, which seeds were soaked in water (Table 2). The biggest fresh weight and 

leaf assimilation area was of shoots, which seeds were soaked in solutions Biojodis 

and Bioforce. It was correspondingly 31.9 and 10.8 %, 19.9 and 21.9 % bigger than 

this of shoots, which seeds were soaked in water. Red beet, which seeds were soaked 

in these solutions, developed faster: they already had third real leaf, while other shoots 

had only two real leaves. 

71

Table 2. Biometrical data of seedlings, which seeds were soaked in growth regulator 

solutions 

2 lentelė. Daržovių daigų, kurių sėklos mirkytos skirtinguose augalų augimo reguliatorių 

tirpaluose, biometrija 

Growth regulator 

Augalų augimo reguliatorius 

Plant height 

Augalo aukštis 

(cm) 

Fresh weight of plant 

Augalo žalioji masė 

(g) 

Leaf assimilation area 

Lapų asimiliacinis plotas 

(cm 2 ) 

1 2 3 4 

Cucumber 

Agurkai ‘Krukiai’ F 1 

Water 

12.11 c* 4.22 c 51.80 c 

Vanduo 

Agronom effect 16.17 e 6.05 f 103.97 f 

Biojodis 9.69 b 3.76 b 53.70 c 

Bioforce 14.30 a 5.74 e 87.36 e 

Biokal 01 9.05 b 2.33 a 41.97 b 

Inzar 12.75 c 5.19 d 98.50 f 

Oksichumat 

5.92 a 1.53 a 24.70 a 

Oksigumatas 

Penergetic p 13.50 c d 5.73 e 68.40 d 

Red beet 

Raudonieji burokėliai ‘Joniai’ 

Water 

12.75 b 1.66 c 35.08 d 

Vanduo 

Agronom effect 15.42 c 0.88 b 22.45 b 

Biojodis 13.05 b 2.19 e 38.88 de 

Bioforce 16.95 d 1.99 d 42.78 f 

Biokal 01 10.62 a 0.83 b 17.73 b 

Inzar 15.54 c 1.09 b 23.98 b 

Oksichumat 

9.67 a 0.59 a 13.51 a 

Oksigumatas 

Penergetic p 14.56 c 1.58 c 28.42 c 

Radish 

Ridikėliai ‘Babtų žara’ 

Water 

14.51 d 2.77 c 87.50 c 

Vanduo 

Agronom effect 14.49 cd 2.45 a 73.31 b 

Biojodis 14.27 cd 2.54 ab 80.83 c 

Bioforce 15.20 e 2.75 c 74.95 b 

Biokal 01 12.27 b 2.25 a 65.08 a 

Inzar 16.22 a 3.48 e 118.10 e 

Oksichumat 

13.65 c 3.25 d 126.60 e 

Oksigumatas 

Penergetic p 15.40 e 3.73 f 111.65 d 

72

1 2 3 4 

Tomato 

Pomidorai ‘Arvaisa’ F 1 

Water 

11.87 c 2.38 d 58.57 d 

Vanduo 

Agronom effect 14.85 f 2.42 d 57.01 d 

Biojodis 11.65 c 2.22 c 68.38 e 

Bioforce 13.40 e 2.48 de 57.85 d 

Biokal 01 7.80 b 1.06 b 42.81 b 

Inzar 12.75 d 2.45 e 42.09 b 

Oksichumat 

6.25 a 0.60 a 29.65 a 

Oksigumatas 

Penergetic p 12.42 d 2.11 c 51.69 c 

* Values indicated by the same letters within the columns are not statistically different at 

P ≤ 0.05 

* Ta pačia raide pažymėtos reikšmės statistiškai nesiskiria, kai P ≤ 0,05 

Radish, which seeds were soaked in the investigated plant growth regulator 

solutions, grew and developed equally in comparison with these, which seeds were 

soaked in water. Radish, which seeds were soaked in solutions Bioforce, Inzar and 

Penergetic p, were correspondingly 4.7 %, 11.8 % and 6.1 % higher than these, which 

seeds were soaked in water (Table 2). 

The biggest fresh weight was of the plants, which seeds were soaked in solution 

Penergetic p. The biggest rootcrop produces plants, which seeds were soaked in solutions 

Biojodis and Inzar (Fig.). 

It was correspondingly 15.1 % and 16.4 % bigger than this of plants, which seeds 

were soaked in water. The biggest leaf assimilation area was of plants, which seeds 

were soaked in solution Oksichumat. It was 44.7 % bigger than this of plants, which 

seeds were soaked in water. 

Agronom effect, Bioforce, Inzar, Penergetic p positively influenced shoot height 

and fresh weight of tomato ‘Arvaisa’ F 1 

, but the biggest leaf assimilation area was of 

tomato shoots, which seeds were soaked in Biojodis solution (Table 2). The height 

of shoots, which seeds were soaked in preparations Agronom effect, Bioforce, Inzar, 

was correspondingly 25.3 %, 13.1 %, 7.6 % and 4.8 % bigger than this of shoots, 

which seeds were soaked in water. Seed soaking in plant growth regulator solutions 

didn’t influence positively tomato leaf assimilation area: it was smaller than this of 

shoots, which seeds were soaked in water (with the exception of shoots, which seeds 

were soaked in Biojodis solution). Leaf assimilation area of shoots, which seeds were 

soaked in Biojodis solution, was 16.7 % bigger than this of shoots, which seeds were 


73

Fig. Root weight (g) of radish, which seeds were soaked in solution of growth regulator: 

1 – water; 2 – Agronom effect; 3 – Biojodis; 4 – Bioforce; 5 – Biokal 01; 6 – Inzar; 

7 – Oksichumat; 8 – Penergetic p. 

Pav. Ridikėlių, kurių sėklos mirkytos įvairiuose augalų 

augimo reguliatorių tirpaluose, šakniavaisio masė, g: 

1 – vanduo; 2 – Agronom effect; 3 – Biojodis; 4 – Bioforse; 

5 – Biokal 01; 6 – Inzar; 7 – Oksigumatas; 8 – Penergetic p. 

Discussion. In order vegetable seeds germinated as evenly and quickly as possible, 

it is necessary correspondingly to prepare them for the sowing. One of the 

methods of seed preparation for sowing is seed soaking. Seeds of some vegetables do 

not germinate for a long time and after soaking them in water they germinate sooner. 

In order to improve vegetable seed germination power and to increase yield, their 

seeds are soaked in plant growth regulator solutions. It was established that soaking 

celery seeds in growth regulators, i. e. Giberelin and others, they germinate sooner 

(Brocklehurst et al., 1982). Tomato ant sweet pepper seeds processed with plant 

growth regulators also germinate sooner and more evenly (Andreoli, Khan, 1999). 

According to the data of Lada and other scientists (2005), when carrot seeds were 

soaked in the solutions of Ambiol, Glycinebetaine and Bioprotect 2 growth regulators 

their germination power increased significantly. The data of our investigation showed 

that not all the investigated plant growth regulators positively influenced vegetable 

seed germination power. Out of the investigated growth regulators, growth regulator 

Bioforce had the the biggest positive influence on seed germination power. When 

seeds were soaked in it germination energy of red beet and raddish seeds increased 

10–30 %. 

Seed soaking in plant growth regulator solutions not only improves their germination 

power, but also shoots became more luxurant, had stronger roots. Plant growth 

74

egulators influence shoot biometrical parameters (Passian, Bennett, 1999). It was 

established that after seed soaking in growth regulator solution or their mixtures, seeds 

not only germinated sooner, but plant overground weight was bigger (Brocklehurst 

et al., 1982). According to the data of Ugur and Kavak (2007), tomato shoots, which 

seeds were soaked in the solutions PP 333 and CCC, were lower. Our investigations 

revealed that not all the growth regulators positively influenced shoot height, leaf 

assimilation area and fresh weight. Some growth regulators positively influenced 

plant height, but didn’t increase plant fresh weight. Agronom effect increased plant 

height and fresh weight of cucumber hybrid ‘Krukiai’ and tomato hybrid ‘Arvaisa’. 

The same regulator increased cucumber shoot leaf assimilation area. Plant growth 

regulator Bioforce increased plant height and fresh weight of red beet and tomato 

hybrid. Biojodis increased red beet plant height and fresh weight. 

Seed soaking in Agronom effect, Bioforce and Penergetic p solutions had the 

biggest influence on cucumber, red beet, tomato and raddish shoot growth and development. 

Conclusions. 1. Plant growth regulator Bioforce had the biggest positive influence 

on red beet and radich seed germination energy. After seed soaking in this regulator, 

germination energy of these vegetable seeds increased 10–30 %. 

2. Cucumber, red beet, tomato and radish seed soaking in Agronom effect, Bioforce 

and Penergetic p solutions had the biggest influence on the growth and development of 

these shoots: Agronom effect increased shoot height and fresh weight of cucumber and 

tomato hybrids and cucumber shoot leaf assimilation area, Bioforce increased shoot 

height and fresh weight of red beet and tomato, Penergetic p increased the height of 

cucumber, radish and red beet and fresh weight of radish was the biggest one. 

Acknowledgement. The research was supported by the Lithuanian State Science 

and Studies Foundation. 

Gauta 2009 06 30 


References 

1. Andreoli C., Khan A. 1999. Matriconditioning integrated with gibberellic acid to 

hasten seed germination and improve stand establishment of pepper and tomato. 

Pesquisa Agropecuária Brasileira, 34(10): 1 953–1 958. 

2. Boehme M., Schevtschenko J., Pinker I. 2005. Effect of biostimulators on growth 

of vegetables in hydroponical systems. Acta Horticulturae, 697: 337–344. 

3. Brigard J. P., Harkess R. L., Baldwin B. S. 2006. Tomato early seedling height 

control using a paclobutrazol seed soak. HortScience, 41(3): 768–772. 

4. Brocklehurst P. A., Rankin W. E. F., Thomas T. H. 1982. Stimulation of celery 

seed germination and seedling growth with combined ethephon, gibberellin and 

polyethylene glycol seed treatments. Plant Growth Regulation, 1(3): 195–202. 

75

5. Bugbee B., White J. W. 1984. Tomato growth as affected by root-zone temperature 

and the addition of gibberellic acid and kinetin to nutrient solutions. Journal 

of the American Society for Horticultural Science, 109: 121–125. 

6. Halter L., Habegger R., Schnitzler W. H. 2005. Gibberellic acid on artichokes 

(Cynara scolymus L.) cultivated in Germany to promote earliness and to increase 

productivity. Acta Horticulturae, 681: 75–82. 

7. Kadiri M., Mukhtar F., Agboola D. A. 1997. Responses of some Nigerian vegetables 

of plant growth regulator treatments. Revista de biologia tropical, 23–28. 

8. Lada R. R., Stiles A., Blake T. J. 2005. The effects of natural and synthetic seed 

preconditioning agents (SPAs) in hastening seedling emergence and enhancing 

yield and quality of processing carrots. Scientia Horticulturae, 106(1): 25–37. 

9. Lopez-Elias J., Salas M. C., Urrestarazu M. Application of indole-3-butyric acid 

by fertigation on pepper plants in soilless culture grown in a greenhouse. Acta 

Horticulturae, 697: 475–479. 

10. Magnitskiy S. V., Pasian C. C., Bennett M. A., Metzger J. D. 2006. Effects of 

soaking cucumber and tomato seeds in paclobutrazol solutions on fruit weight, 

fruit size, and paclobutrazol level in fruits. HortScience, 41 (6): 1 446–1 448. 

11. Panajotov N. D. 1997. The effect of plant growth regulator atonic on the yield 

and quality of the reproduced seeds of sweet pepper. Acta Horticulturae, 462: 

757–762. 

12. Papadopoulos A. P., Saha U., Hao X., Khosla S. 2006. Response of rockwool-grown 

greenhouse cucumber, tomato, and pepper to kinetin foliar sprays. 

HortTechnology, 16(3): 32–35. 

13. Pasian C. C., Bennett M. A. 1999. Seed coats as plant growth regulator carriers 

in bedding plant production. Acta Horticulturae, 504: 93–98. 

14. Saglam N., Gebologlu N., Yilmaz E., Brohi A. 2002. The effects of different 

plant growth regulators and foliar fertilizers on yield and quality of crisp lettuce, 

spinach and pole bean. Acta Horticulturae, 579: 619–623. 


analizė taikant kompiuterines programas ANOVA, STAT, SPLIT-PLOT iš paketo 

SELEKCIJA ir IRRISTAT. Akademija. Kėdainių r. 

16. Ugur A., Kavak S. 2007. The effects of PP 333 and CCC on seed germination 

and seedling height control of tomato. Acta Horticulturae, 729: 205–208. 

17. Гущина Л., Къдрев Е. 1987. Ускоряване на узряването и добива на ранни 

домати. Физиология растений, 13(2): 82–87. 

76


Augimo reguliatorių įtaka daržovių sėklų dygimo energijai ir 

daigų vystymuisi 

J. Jankauskienė, E. Survilienė 

Summary 

2007 metais Lietuvos sodininkystės ir daržininkystės institute tirta augalų augimo reguliatorių 

įtaka daržovių sėklų dygimo energijai ir biometrijai. Agurkų ‘Krukiai’ F 1 

, burokėlių ‘Joniai’, 

ridikėlių ‘Babtų žara’ ir pomidorų ‘Arvaisa’ F 1 

sėklos mirkytos augimo reguliatorių Biojodis, 

Biokal 01, Bioforse, Agronom effect, Inzar, Oksigumatas, Penergetic p tirpaluose. Kontroliniame 

variante sėklos mirkytos vandenyje. Po mirkymo sėklos pasėtos į polimerines kasetes, kuriose 

augalai auginti 30 dienų šiltnamyje. Nustatyta sėklų dygimo energija, atlikti daigų biometriniai 

matavimai (augalų aukštis, masė, lapų skaičius, asimiliacinis plotas). Augalų augimo reguliatoriai 

Oksigumatas, Agronom effect, Bioforse bei Penergetic p darė teigiamą įtaką ridikėlių, pomidorų 

sėklų dygimo energijai bei agurkų, burokėlių, pomidorų ir ridikėlių daigų augimui bei vystymuisi. 

Reikšminiai žodžiai: agurkai, augimo reguliatoriai, biometrija, burokėliai, daigai, dygimo 

energija, pomidorai, ridikėliai. 

77




Powdery mildew of strawberries in Latvia under field 

conditions 

Svetlana Jarmoliča, Biruta Bankina 

Institute of Soil and Plant Sciences, Latvia University of Agriculture, Liela Street 2, 

Jelgava, LV-3004, e-mail: Biruta.Bankina@llu.lv; lanasvet2@inbox.lv 

Mildew of strawberries is a widespread disease over the world, but it was not detected 

in Latvia under open field conditions. Mildew of strawberries was noted first in open field in 

Latvia in 2007, but regular observations were started in Research and Study farm “Vecauce” 

of LLU in 2008. 

Observations were carried out in different varieties in strawberry plantations of different 

age. 

Mildew was determined only in two varieties – ‘Zefyr’ and ‘Kokinskaja rannaja’. 

Incidence of the disease fluctuated from 9–15 % depending on the age of a strawberry plantation 

and was higher in 3-year-old plantations. Severity of the disease was not high and did not 

reach 1 point (evaluation scale 0–5 points). 

Morphological properties of Podosphaera (chasmothecia and conidia) were described. 

We suggest, that mildew of strawberries in Latvia was caused by fungus from the genera 

Podosphaera (former Sphaerotheca), but more detailed investigations are necessary. The 

main aim of the research was to estimate harmfulness of strawberry powdery mildew and to 

clarify life cycle of pathogen in Latvia under field conditions. 

Key words: diagnostic, diseases, Podosphaera, Sphaerotheca. 

Introduction. Mildew of strawberries is a widespread disease over the world. 

Powdery mildew of strawberries was noted only in glasshouses, but was not detected 

in Latvia under field conditions. There are some possible reasons for emergence of 

powdery mildew: new varieties and climatic changes. Milder winters might allow 

overwintering of Podosphaera spp. and hot summers increase rate of disease progress 

(Boland et al., 2004). 

The disease damages all aerial plant tissues, including fruits (Maas, 1998). 

Systematic of powdery mildew causal agents sharply changed during the last years 

(Glawe, 2008). The taxonomy of Erysiphales recently was revised basing on DNA 

sequence data. Identification pathogens from Erysiphales now require morphology 

peculiarities of teleomorph and anamorph, incorporates characteristics to the whole 

79

fungus (anamorph plus teleomorph, i. e. the holomorph). Powdery mildew genera are 

now grouped into five tribes (Heffer et al., 2006). 

Podosphaera aphanis and Podosphaera macularis were described as causal agents 

of powdery mildew. Formerly Sphaerotheca macularis was thought to be pathogen 

of strawberries. 

Earlier literature findings do not distinguish clearly between the P. aphanis and 

P. macularis, but P. aphanis is the most important causal agent of powdery mildew 

in UK (Hal et al., 2008). 

DNA analyses of an isolate of the synonymous species Sphaerotheca aphanis 

from strawberry were 100 % identical to P. aphanis (Pertot et al., 2007). 

Severity of powdery mildew development depends on meteorological factors: 

temperature (the optimum 15–25 °C) and high relative humidity – higher than 75 %, 

but less than 98 % (Amsalem et al., 2006). Developing of mycelia in green tissue was 

observed under all conditions, where the pathogen survived irrespective of visible 

symptoms, but sporulation was observed from 5–30 °C (Miller et al., 2003). 

The main source of infection is chasmothecia (formerly cleistothecia), and time 

of ripening and liberation of ascospores is important to development of integrated 

control of powdery mildew. Viable ascospores in the chasmothecia were found from 

April till June in Norway (Gadoury et al., 2007). 

The central aim of research was to estimate harmfulness of strawberry powdery 

mildew and clarify life cycle of pathogen in Latvia under field conditions. 

Object, methods and conditions. Regular observations were carried out in Research 

and Study farm “Vecauce” of LLU in 2008. 

Different strawberry varieties (‘Fratina’, ‘Pegasus’, ‘Polka’, ‘Pandora’, ‘Zefyr’, 

‘Honeoye’, ‘Kokinskaja rannaja’), 2-year and 3-year old plantations were evaluated. 

Incidence and the severity of powdery mildew were noted in autumn. Incidence 

of the disease was expressed in percent, but severity in points. A scale of 0–5 was 

developed to evaluate the severity of mildew: 0 – healthy plant; 1 – first symptoms of 

the disease; 2 – infected 2–3 leaves; 3 – infected 4–5 leaves; 4 – infected more than 

5 leaves; infected all plant. 

Morphological peculiarities of fruit bodies of pathogen were studied and described 

under microscope at the Institute of Soil and Plant Sciences. Identification of causal 

agent of strawberry powdery mildew was performed. 

Results. Powdery mildew was determined only in two varieties – ‘Zefyr’ and 

‘Kokinskaja rannaja’ – in the autumn of 2008. 

Incidence of the disease fluctuated from 9–15 %, depending on the age of strawberry 

plantation, being higher in 3-year-old plantation (Fig. 1). However, severity of 

the disease was not high and did not achieve 1 point. 

80

Fig. 1. Incidence of powdery mildew depending on variety and age of plantation 

1 pav. Netikrosios miltligės paplitimas, priklausomai nuo veislės ir plantacijos amžiaus 

Morphological properties of chasmothecia (former cleistothecium) and conidia 

were described in the autumn of 2008 (Figs. 2 and 3). We suggest that powdery mildew 

of strawberries in Latvia was caused by fungus from the genera Podosphaera (former 

Sphaerotheca). 

Fig. 2. Chasmothecia with ascus and asco spores of Podosphaera spp. 

2 pav. Chasmothecia su Podosphaera spp. askosporomis ir askais 

Fig. 3. True conidia chains of pathogen 

3 pav. Tikrosios ligos sukėlėjo konidijų grandinės 

81

Discussion. In Latvia, powdery mildew of strawberries was for the first time 

noted in field in 2007, but this disease has been described all over the world. There 

are no data available with regard to resistance of strawberry varieties under conditions 

of Latvia, but different level of resistance was observed. 

Identification of species was not done now; more detailed investigations are necessary. 

Genera Podosphaera were established as causal agent of strawberry mildew, 

related with a new system of pathogen systematic, but most of authors used previous 

name Sphaerotheca. 

Chasmothecia is a spherical fruiting body without natural opening. Morphological 

peculiarities of chasmothecia appendages, number of asci and production of conidia (a 

single conidium, true chain or pseudochain) are the most important factors for identification 

of causal agent of mildew. Chasmothecia of Podosphaera contain one single 

ascus, appendages are hypha-like or branched and conidia form true chains (Heffer 

et al., 2006). Conidia in chains are ellipsoidal to barrel-shaped as reported by others 

researchers (Santos et al., 2002). Fig. 2 and Fig. 3 show chasmothecia with a single 

ascus, hypha-like appendages and true chains of conidia. 

Investigation of peculiarities of the disease life cycle is a very important task. 

Control scheme of mildew could be based on incidence of chasmothecia (Berrie et al., 

2002). Viable asco spores were detected in Latvia in autumn; investigations are continued. 

Similar investigations were carried out in Norway, asco spores were found in 

spring (Gadoury et al., 2007). It means that the same situation is possible in Latvia. 

However, season most favourable for infection is unclear in Latvia yet. 

Conclusions. Powdery mildew of strawberries was first described in Latvia in 

the autumn of 2008; genera of causal agent were identified as Podosphaera. 

Further investigations are necessary for precise identification of causal agent and 

clarification of pathogen life cycle under conditions of Latvia. 

Gauta 2009 06 30 


References 

1. Amsalem L., Freeman S., Rav-David D., Nitzani J., Sztejnberg A., Pertot I., 

Elad Y. 2006. Effect of Climatic Factors on Powdery Mildew Caused by 

Sphaerotheca macularis f. sp. Fragariae on Strawberry. European Journal of 

Plant Pathology, 114(3): 283–292. 

2. Berrie A. M., Harris D. C., Xu X. M. 2002. A potential system for managing 

Botrytis and powdery mildew in main season strawberries. Acta Horticulture, 

567: 647–649. 

3. Boland G. J., Melzer M. S., Hopkin A., Higgins V., Nassuth A. 2004. Climate 

change and plant diseases in Ontario. Canadian Journal of Plant Pathology, 26: 

335–350. 

82

4. Gadoury D. M., Stensvand A., Seem R. C., Heidenreich C., Herrero M. L., 

Welser M. 2007. Overwinter survival of cleistothecia, ascospore release, and 

infection of strawberry by Podosphaera macularis in New York and Norway. 

Phytopathology, 97: S38. 

5. Glawe A. 2008. The Powdery Mildews: A Review of the World’s Most Familiar 

(Yet Poorly Known) Plant Pathogens. Annual Review of Phytopathology, 46: 

27–51. 

6. Hall A. M., Dodgson J. L. A., Farooq M. 2008. Comparison of Podosphaera 

macularis and P. aphanis and the role of chasmothecia on strawberries. Journal 

of Plant Pathology, 90(2): S2.161. 

7. Heffer V., Johnson K. B., Powelson M. L., Shishkoff N. 2006. Identification of 

powdery mildew fungi. The Plant Health Instructor. DOI:10.1094/PHI-I-2006- 

0706-01 

8. Maas J. L. 1998. Compendium of strawberry diseases. 2 nd ed. American 

Phytopathological Society Press, St. Paul. Minn. 

9. Miller T. V., Gubler W. D., Geng S., Rizzo D. M. 2003. Effects of temperature 

and water vapor pressure on conidial germination and lesion expansion of 

Sphaerotheca macularis f. sp. fragariae. Plant diseases, 87(5): 484–492. 

10. Pertot I., Fiamingo F., Amsalem L., Maymon M., Freeman S., Gobbin D., Elad 

Y. 2007. Sensitivity of two Podosphaera aphanis populations to disease control 

agents. Journal of Plant Pathology, 89(1): 85–96. 

11. Santos B., Blanco C., Porras M., Barrau C., Romero F. 2002. First Confirmation 

of Sphaerotheca macularis on Strawberry Plants in Southwestern Spain. Plant 

Disease, 86(9): 1 049. 


Braškių netikroji miltligė Latvijoje lauko sąlygomis 

S. Jarmoliča, B. Bankina 

Santrauka 

Braškių miltligė – visame pasaulyje paplitusi liga, tačiau Latvijoje lauko sąlygomis ji dar 

nebuvo aptikta. Pirmą kartą atvirame lauke Latvijoje ji pastebėta 2007 metais, o reguliarūs 

stebėjimai pradėti LLU tyrimų ir studijų ūkyje “Vecauce” 2008 metais. 

Stebėtos skirtingos braškių veislės įvairaus amžiaus plantacijose. Miltligė aptikta tik 

dviejose veislėse – ‘Zefyr’ ir ‘Kokinskaja rannaja’. Ligos paplitimas svyravo 9–15 %, priklausomai 

nuo braškių plantacijos amžiaus, ir buvo didesnis 3 metų amžiaus plantacijose. Ligos 

žalingumas nebuvo didelis ir nesiekė 1 balo (vertinant pagal 0–5 balų skalę). 

Buvo apibūdintos morfologinės savybės Podosphaera (chasmothecia ir konidijų). Spėjame, 

kad braškių miltligę Latvijoje sukėlė Podosphaera (buvusios Sphaerotheca) genties grybas, 

tačiau tam patvirtinti reikalingi nuodugnesni tyrimai. Pagrindinis šių tyrimų tikslas buvo 

įvertinti braškių netikrosios miltligės kenksmingumą ir išsiaiškinti ligos sukėlėjo vystimosi 

ciklą Latvijoje lauko sąlygomis. 

Reikšminiai žodžiai: ligos, nustatymas, Podosphaera, Sphaerotheca. 

83




Influence of neemazal-T/S on Mamestra brassicae L. 

Katrin Jõgar*, Luule Metspalu, Külli Hiiesaar, Liina Loorits, 

Angela Ploomi, Aare Kuusik and Anne Luik 

Institute of Agricultural and Environmental Sciences, Estonian University of Life 

Sciences, 1 Kreutzwaldi St., 51014 Tartu, Estonia, e-mail katrin.jogar@emu.ee 

The aim of this study was to explore the effects of the botanical insecticide NeemAzal- 

T/S on Mamestra brassicae, known an important cabbage pest. The experiments were carried 

out in the experimental garden of the Estonian University of Life Sciences in the summer 

of 2006. During the experiment the effect of different concentrations and treating methods 

of the preparation (0.03 % and 0.3 % solution spraying and 0.3 % solution watering) were 

monitored. 

During the observation period M. brassicae were found in lower numbers from treated 

plants than from untreated plants. A comparison of treated variants with control revealed 

statistically significant differences in the number of M. brassicae. There were no significant 

differences between the treated variants. Seasonal dynamics of M. brassicae showed that the 

population peak was in the beginning of July and after that the number of pests started to 

decrease. Spraying the cabbage with NeemAzal-T/S 0.3 % solution decreased the inhabitation 

by cabbage moth. The effect was not as clear in the other treatments. NeemAzal-T/S acted on 

cabbage moth females as a weak repellent and oviposition deterrent. According to our results, 

0.3 % concentration of NeemAzal-T/S was most effective against cabbage moth and spraying 

was found to be more effective than watering. 

Key words: biopesticide, Mamestra brassicae, oviposition preference. 

Introduction. Cabbage moth, Mamestra brassicae L. is a highly polyphagous 

species, particularly associated with cruciferous crops (Bretherton et al., 1979), but 

also feeding on a wide range of other plant species (Turnock, Carl, 1995). In Estonia 

M. brassicae is one of the most significant pests of cruciferous crops, but is also 

widely known pest throughout North West Europe. In various climatic zones cabbage 

moth can produce a number of generations during summer; in Estonia it is mainly a 

univoltine species, hibernating as diapausing pupae in the soil. Under our conditions 

synthetic insecticides are dominant in plant protection strategies against cabbage 

moth and botanical insecticides are not very common in practice. 

Botanical insecticides are useful and desirable tools in most pest management 

programs because they can be effective and often complement the actions of natural enemies 

(Ascher, 1993; Schmutterer, 1995). The main advantages of using bioinsecticide 

85

are reduced toxicity to humans, rapid and complete degradation in the environment, 

low risk for resistance and selective properties for non-target organisms, including 

natural enemies of pest and insects-pollinators (Hoelmer et al., 1990; McCloskey et al., 

1993; Nauman et al., 1994; Schmutterer, 1995). 

Botanical insecticides are less harmful than synthetic ones and affect many insects 

in different ways (Schmutterer, 1995; Villanueva-Jiménez et al., 2000; Metspalu et al., 

2001; Durmusoglu et al., 2003). Among natural pesticides the compounds from neem 

tree (Azadirachta indica A. Juss, Meliaceae) have a number of properties useful for 

insect pest management. Neem extracts are widely used around the world either singly 

in Integrated Pest Management or in conjunction with synthetic pesticides (for review 

see Mordue (Luntz), Nisbet, 2000). Neem-based insecticides containing different 

compounds have been reported to control more than 400 species of insects, including 

important pests, such as leafminers, aphids and whiteflies (Isman, 1999; Walter, 1999). 

Azadirachtin, a steroid-like tetranortriterpenoid derived from neem trees, is a strong 

anti-feedant, repellent, growth regulator, molting inhibitor, sterilant and oviposition 

deterrent for a wide variety of phytophagous insects (Koul, 1992; Schmutterer, 1995; 

Mordue (Luntz), 2004). Azadirachtin has also shown direct detrimental and histopatological 

effects on most insect tissues, e. g. muscles, body fat, and gut epithelial cells 

(Mordue (Luntz), 2004). 

The action of neem depends on the pest species, the time of application, treating 

methods and the concentration used. It is also important to determine the efficacy of 

neem in different regions of the world. One preparation may act differently with different 

populations. The aim of this study was to elucidate the actions and efficiency 

of different concentrations of commercial botanical insecticide NeemAzal-T/S on the 

cabbage moth, Mamestra brassicae, under field conditions in Estonia. 

Object, methods and conditions. The experiments were carried out in the experimental 

garden of the Estonian University of Life Sciences in the summer of 2006. In 

the present experiments NeemAzal-T/S (1 % Azadirachtin) (from Trifolio-M GmbH, 

Germany), was used. There were four variants: two different concentrations of NeemAzal-T/S 

– 0.03 % and 0.3 % were used for spraying and 0.3 % concentration of 

Neem Azal T/S was used for watering of testing plots. 

White cabbage (Brassica oleracea (L.) var. capitata f. alba) plants were grown 

from seed, kept in glasshouse until they reached the 3 true leaf stage. In mid-May 

plants were replanted to the experimental field. Each variant consisted of 9 plants per 

plot (three rows of three plants spaced at 70 cm intervals). All variants had three replications. 

To prevent larvae from leaving the plots, a 20 cm wide strip of dill (Anethum 

graveolens L.), which is not a food plant of M. brassicae larvae, was sown around 

each plot. Larvae of M. brassicae on all the plots were sampled at 7-day intervals from 

4 July to 12 September. Eggs and larvae were removed from plants by hand picking, 

to avoid repeated counting. The first spraying and watering with NeemAzal-T/S was 

made after the first counting (4 July) of M. brassicae eggs and larvae. The treatments 

were applied at weekly intervals during the entire observing period. 

Tests were performed using the statistic package StatSoft ver. 7, Inc./USA. Data 

have been presented as mean ± standard error. Statistical comparisons were performed 

with repeated measures ANOVA by the Ficher LSD test. All means were considered 

significantly different at the p < 0.05 level. 

86

Results. A comparison of treated variants with control revealed statistically significant 

differences in the number of M. brassicae larvae (ANOVA F 3,18 

=12.30; 

df = 3; p < 0.05; LSD test p < 0.05). The number of M. brassicae was reliably lower 

in all treated variant (p < 0.05) (Fig. 1) during the whole observation period. Our results 

showed that butterflies preferred the untreated plants as the site for oviposition, 

37.8 % of caterpillars counted during the whole observation period were gathered 

from this variant. 

Fig. 1. Mean number of cabbage moth (Mamestra brassicae L.) 

individuals on NeemAzal-T/S treated variants. Columns with different letters are 

significantly different (Ficher LSD test p < 0.05). 

1 pav. Kopūstinio pelėdgalvio (Mamestra brassicae L.) vidutinis gausumas 

nimazaliu apdorotuose variantuose. Stulpeliai pažymėti skirtingomis 

raidėmis patikimai skiriasi (Fišerio testas, kai p < 0,05). 

Statistical analysis of the results indicated that there were no significant differences 

between the treated variants (LSD test, p > 0.05), although there existed a considerable 

tendency in the direction of lower number of M. brassicae in the variant treated 

with 0.3 % neem. The comparison of the differently treated variants revealed that the 

cabbage moth selected least the plots sprayed with 0.3 % neem preparation (16.7 %), 

followed by sprayed with 0.03 % neem (21.2 %) as the site for oviposition and the 

third choice was watered variant (24.3 %). 

The first observation before treatments revealed high infestation of cabbage moth 

in all variants; there were no significant differences between variants (Fig. 2). One 

week after treatments (11.07) the number larvae fall drastically in all treated variants; 

the lowest number of larvae was found on plots sprayed with 0.3 % neem preparation. 

Comparison of number of M. brassicae in all test variants revealed significant 

difference with control (p = 0.02), but there were no differences between treatments 

(p > 0.05; Fig. 2). 

87

Fig. 2. Seasonal abundance of Mamestra brassicae on differently NeemAzal-T/S 

treated variants (mean ± SE). Columns with asterisks are significantly different 

from control (Tukey test p < 0.05). 

2 pav. Mamestra brassicae sezoninis gausumas skirtingai nimazaliu apdorotuose 

variantuose (vidurkis ± SE). Stulpeliai, pažymėti žvaigždutėmis, 

patikimai skiriasi nuo kontrolinio varianto (Tukey testas p < 0,05). 

Only few larvae were found at the third observation (18.07) on 0.3 % sprayed 

variant; somewhat more – on sprayed with 0.03 %, followed by watered variant. In 

control the number of pest was declined slightly. The same trend continued in all 

variants till the end of observations. 

Discussion. Cabbage moth mainly selects an oviposition site by odour cue, whereas 

the search process is, to some extent, influenced by visual cues (Rojas et al., 

2000). Data from literature reveal that neem is useful as an ovipositional repellent for 

polyphagous pests (Larew, 1990; Liu, Stansly, 1995). According to Metspalu et al., 

(2001) results NeemAzal-T/S had a repellent effect on the adults of large white butterfly 

(Pieris brassicae). Similar results were found in Facknath (1998) – different 

neem preparations had a repellent effect on Plutella xylostella adults. Meadow et al., 

(2001) found a repellent effect of neem on oviposition of turnip root fly (Delia floralis) 

and cabbage moth. Our data showed that cabbage moth oviposition was reduced 

on cabbage treated with NeemAzal-T/S in comparisson with control. Probably the 

neem-treating misinformed the M. brassicae adults and they did not find the plants 

or they were not able to lay the eggs. Especially decreased the inhabitation by the 

88

cabbage moth in variant with 0.3 % concentration. Similarly to our observations, 

the repellent effect of neem extracts against M. brassicae oviposition was found in 

a study by Seljasen and Meadow (2006). Naumann and Isman (1995) found a similar 

reduction in oviposition by the noctuid moth Spodoptera litura on neem-treated 

plants. Neem-based insecticides deter oviposition by some other lepidopteran, and 

some homopteran, coleopteran and dipteran pests (Saxena, 1989; Schmutterer, 1995; 

Butler et al., 1991; Singh, Singh, 1998; Akey, Henneberry, 1999). 

The present research showed that direct contact with the Neem Azal T/S decreased 

M. brassicae egg survival and larvae that fed on neem treated leaves had lower 

survivorship. During the observation we found both dead egg clusters and larvae from 

neem-treated plants. According to Liang et al. (2003) results Agroneem, Ecozin and 

Neemix had lethal effects on the diamondback moth larvae, and neem oil reduced egg 

hatching and larval survival in larvae of Helicoverpa armigera (Ma et al., 2000). 

Our tests showed that the lower number of M. brassicae on neem treated plants; 

that could have resulted from direct toxicity of the neem extract. Respectively to 

Mancebo et al. (2002) data, Azatin, neem seed extract containing 3 % azadirachtin, 

has been reported to cause quick direct toxicity in the mahogany shootborer, Hypsipyla 

grandella larvae. 

According to our results, 0.3 % concentration of NeemAzal-T/S was most effective 

against cabbage moth and spraying was found to be more effective than watering. 

Systemic transport of neem in plants and controlling effects are also documented for 

the cabbage pest P. brassica (Osman, Port, 1990) and Plutella xylostella (Wendorf, 

Shüler, 1992). Our results confirm previous statements. 

Conclusion. From our results it can be concluded that under our conditions it is 

possible to control M. brassicae with NeemAzal-T/S. 

Acknowledgements. This research was supported by Grants No 6722 and 7130 

of the Estonian Science Foundation, and Estonian Ministry of Education and Research 

targeted financing project No. SF 0170057s09. 

Gauta 2009 06 30 


References 

1. Akey D. H., Henneberry T. J. 1999. Control of silverleaf whitefly with neem 

product azadirachtin as Bollwhip TM in upland cotton in Arizona. Proceedings of 

Beltwide Cotton Conferences, National Cotton Council of America, Memphis, 

TN, 914–917. 

2. Ascher K. R. S. 1993. Nonconventional insecticidal effects of pesticides available 

from the neem tree, Azadirachta indica. Archives of Insect Biochemistry and 

Physiology, 22: 433–449. 

89

3. Bretherton R. F., Goater B., Lorime R. I. 1979. Noctuidae. In: J. Heath and 

A. M. Emmet (eds.) The Moths and Butterflies of Great Britain and Ireland. 

Curwen Booka, London, 120–278. 

4. Butler G. D. Jr., Puri S. N., Henneberry T. J. 1991. Plant-derived oil and detergents 

solutions as control agents for Bemisia tabaci and Aphis gossypii on 

cotton. Southwestern Entomologist, 16: 331–337. 

5. Durmusoglu E., Karsavuran Y., Ozgen I., Guncan A. 2003. Effects of two different 

neem products on different stages of Nezara viridula (L.) (Heteroptera, 

Pentatomidae). Journal of Pest Science, 76: 151–154. 

6. Facknath S. 1998. Integrated pest management of Plutella xylostella an important 

pest of crucifers in Mauritius. (www.uom.ac.mu/Faculties/foa/AIS/ 

SIROIWEBUK/maurice/farc/amas97/p13txt.htm) 

7. Hoelmer K. A., Obsorne L. S., Yokomi R. K. 1990. Effects of neem extracts on 

beneficial insects in greenhouse culture. In: J. C. Lovke and R. H. Lawson (eds.) 

Neem-potential in pest management programs. USDA Agricultural Research 

Service, 86, 100–105. 

8. Isman M. B. 1999. Neem and related natural products. In: F. R. Hall and 

J. J. Menn (eds.) Biopesticides: Use and Delivery. Humana Press, Totowa, NJ. 

139–154. 

9. Koul O. 1992. Neem alleochemicals and insect control. In: Rizvi, R. S. J. H. and 

Rizvi, R.H. (eds.) Allelopathy; basic and applied aspects, 86, 100–105. 

10. Larew H. G. 1990. Activity of neem seed oil against greenhouse pests. In UCDA 

Neem Workshop, USDA-ARG 86, 128–131. 

11. Liang G. M., Chen W., Liu T. X. 2003. Effects of tree neem-based insecticides on 

diamondback moth (Lepidoptera: Plutellidae). Crop Protection, 22: 333–340. 

12. Liu T. X., Stansly P. A. 1995. Toxicity and repellency of some biorational 

Insecticides to Bemisia argentifolii on tomato plants. Entomologia Expermentalis 

et Applicata, 74: 137–143. 

13. Ma D. L., Gordh G., Zalucki M. P. 2000. Biological effects of azadirachtin on 

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) fed on cotton and artificial 

diet. Australian Journal of Entomology, 39: 301–304. 

14. Mancebo F., Hilje G. A. M., Salazar R. 2002. Biological activity of two neem 

(Azadirachta indica (A. Juss., Meliaceae) products on Hypsipyla grandella 

(Lepidoptera: Pyralidae) larvae. Crop Protection, 21: 107–112. 

15. McCloskey C., Arnason J., Donskov N., Chenier R., Kaminski J., Philogene B. 

J. R. 1993. Third troffic level effects of azadirachtin. Canadian Entomologist, 

125: 163–165. 

16. Meadow R., Seljasen R., Brynildsen P. 2001. The effects of neem extracts on the 

turnip root fly and the cabbage moth. Practice oriented results of the use of plant 

extracts and pheromones in pest control. Proceedings of the IX Workshop, (eds. 

H. Kleeberg & C. P. W. Zebitz), 53–60. 

17. Metspalu L., Luik A., Hiiesaar K., Kuusik A., Sibul, I. 2001. On the influence 

of Neem Preparations on some agricultural and Forest Pests. Practice oriented 

results of the use of plant extracts and pheromones in pest control. Proceedings 

of the IX Workshop, (eds. Kleeberg, H. & Zebitz, C. P. W.), 95–103. 

90

18. Mordue (Luntz) A. J. 2004. Present Concepts of the Mode of Action of 

Azadirachtin from Neem. Neem: Today and in the New Millennium. (eds. 

O. Koul & S. Wahab) Kluwer Academic Publishers, London, 229–243. 

19. Mordue (Luntz) A. J., Nisbet A .J. 2000. Azadirachtin from the neem tree A. indica: 

its action against insects. Anais da Sociedate Entomologica do Brasil, 29(4): 

615–632. 

20. Nauman K., Currie R. W., Isman B. M. 1994. Evaluation of the repellent effect 

of a neem insecticide on foraging honey bees and other pollinators. Canadian 

Entomologist, 126: 225–230. 

21. Naumann K., Isman M. B. 1995. Evaluation of neem Azadirachta indica seed 

extracts and oils as oviposition deterrents to noctuid moths. Entomologia 

Expermentalis et Applicata, 76: 115–120. 

22. Osman M. Z., Port R. G. 1990. Systemic action of neem seed substances against 

Pieris brassica. Entomologia Expermentalis et Applicata, 54: 297–300. 

23. Rojas J. C., Wyatt T. D., Birch M. C. 2000. Flight and oviposition behaviour 

toward different host plant species by the cabbage moth, Mamestra brassicae 

(L.) (Lepidoptera: Noctuidae). Journal of Insect Behavior, 13(2): 297–300. 

24. Saxena R. C. 1989. Insecticides from neem. In: J. T. Arnason, B. J. R. Philogene, 

P. Morand (eds.). Insecticides of Plant Origin, American Chemical Society, 

Washington, DC, 110–135. 

25. Schmutterer H. 1995. The neem tree: source of unique natural products for 

integrated pest management, medicine, industry and other purposes. VCH 

Publications, Weinheim, Germany. 

26. Seljasen R., Meadow R. 2006. Effects of neem on oviposition and egg and larval 

development of Mamestra brassicae L.: Dose response, residual activity, 

repellent effect and systemic activity in cabbage plants. Crop Protecion, 25: 

338–345. 

27. Singh S., Singh R. P. 1998. Neem (Azadirachta indica) seed kernel extracts and 

azadirachtin as oviposition deterrents against the melon fly (Bactrocera cucurbitae) 

and oriental fruit fly (Bactrocera dorsalis). Phytoparasitica, 26: 1–7. 

28. Turnock W. J., Carl K. P. 1995. Evaluation of palearctic Eurithia consobrina 

(Diptera: Tachinidae) as a potential biocontrol agent for Mamestra configurata 

(Lepidoptera, Noctuidae) in Canada. Biocontrol Science and Technology, 

5: 55–67. 

29. Villanueva-Jiménez J. A., Hoy M. A., Davies F. S. 2000. Field evaluation of 

integrated pest management-compatible pesticides for the citrus leafminer 

Phyllocnistis citrella (Lepidoptera: Gracillariidae) and its parasitoid Ageniaspis 

citricola (Hymenoptera: Encyrtidae). Journal of Economic Entomology, 93: 

357–367. 

30. Walter J. F. 1999. Commercial experience with neem products. In: F. R. Hall, 

J. J. Menn (eds.) Biopesticides: Use and delivery. Humana Press, Totowa, NJ, 

139–154. 

91

31. Wendorf M., Shüler C. 1992. Versuch über die systemische Wirkung von 

Neemenextracten auf Plutella xylostella. Proceedings of the First Workshop: 

Practise Oriented Results on Use and Production of Neem-Ingredients (ed. 

H. Kleeberg). 19 th –20 th June 1992. Wetzlar, Germany, Trifolio-M-GmbH, 

Lahnau, 45–51. 


Nimazalio T/S poveikis Mamestra brassicae L. 

K. Jõgar*, L. Metspalu, K. Hiiesaar, L. Loorits, A. Ploomi, 

A. Kuusik, A. Luik 

Santrauka 

Tyrimo tikslas buvo ištirti botaninio insekticido nimazalio T/S poveikį žalingam 

kopūstų kenkėjui Mamestra brassicae. Tyrimai buvo atlikti Estijos gamtos mokslų universiteto 

ekspermentiniame darže 2006 m. vasarą. Buvo tirtas preparato skirtingų koncentracijų 

ir apdorojimo būdų (0,03 % ir 0,3 % tirpalo purškimo ir 0,3 % tirpalo laistymo) poveikis. 

Stebėjimo metu mažiau M. brassicae buvo ant apdorotų augalų nei ant neapdorotų. 

Variantų palyginimas parodė patikimus M. brassicae skirtumus tarp apdorotų variantų 

ir kontrolės. Tarp apdorotų variantų patikimų skirtumų nerasta. Sezoninis M. brassicae 

kitimas rodo, kad gausiausiai šių kenkėjų buvo liepos pradžioje, po to jų skaičius 

pradėjo mažėti. Purškiant kenkėjus nimazalio T/S 0,3 % tirpalu sumažėjo kopūstinių 

pelėdgalvių gyvenimo trukmė. Poveikis kituose variantuose nebuvo ryškus. Nimazalis T/S 

veikė kopūstinių pelėdgalvių pateles kaip silpnas repelentas ir šiek tiek atbaidydavo nuo 

kiaušinių dėjimo. Remiantis mūsų rezultatais, 0,3 % nimazalio T/S koncentracija buvo 

efektyviausia nuo kopūstinių pelėdgalvių ir purškimas buvo efektyvesnis nei laistymas. 

Reikšminiai žodžiai: biopesticidas, kiaušinių dėjimo pirmenybė, Mamestra brassicae. 

92




Sensitivity of Venturia inaequalis populations to 

krezoxym methyl 

Veranika Kamardzina 

RUC “Institute of Plant Protection” p. Priluki, Minsk region, Belarus, 

e-mail belizr@tut.by 

As a result of investigations carried out in the industrial orchards in 2000–2006, significant 

decrease of sensitivity of apple scab agent Venturia inaequalis populations to krezoxym methyl 

(Strobi) from strobilurine group was determined. In orchard “Kletsky”, Minsk district, in the 

first year of the application of this fungicide the amount of sensitive isolates has made 100 % 

and the resistance factor has not increased 24.2. After 4 times of preparation application in 

2001 a proportion of sensitive isolates decreased up to 80.2 % and the population resistance 

factor increased 2.6 times. When Strobi application was stopped in 2002, a tendency to sensitivity 

decrease has remained: a proportion of sensitive isolates decreased up to 63.3 % and 

the resistance factor reached 88.4. 

In orchard “Rassvet”, Brest district, where krezoxym methyl was started to apply since 

1998, in 2005 a proportion of the resistant isolates has made 59.2 %, and the population 

resistance factor was 117.2. When the number of treatments decreased up to one in 2006, 

the situation has not changed essentially, what proves the progressive loss of V. inaequalis 

population sensitivity to the fungicide. 

Key words: apple-tree, effective concentration (ЕС 50 

), fungicides, industrial orchards, 

resistance, strobilurin, Venturia inaequalis. 

Introduction. Apple-tree is the basic fruit crop in Belarus occupying 95 % of the 

total area of large-and-small fruit plantations (Самусь, 2008). When there was passed 

to the intensive gardening in the Republic, apple scab (the agent – fungus Venturia 

inaequalis (Coockе) Winter) development in the industrial apple orchards practically 

annually reaches epiphytoty. Thus, crop losses on susceptible to the disease varieties 

reach 60 %. Under the conditions of Belarus for orchard protection against scab the 

limited assortment of systemic fungicides is used. To get a standard production, such 

preparations as Score EC from triazole group; Chorus WDG from anilino-pirimidine 

group and Strobi, 500 g kg -1 from strobilurine group are the most often applied. Long 

application and repeated treatments by these fungicides promote accumulation of 

resistant forms in a phytopathogen population. 

Fungicides from strobilurine group have appeared on the market of Belarus in 

the late nineties. Strobilurines are the analogues of a natural antibiotic strobilurine 

93

A, which is produced by fungi Strobilurus tenacellur and Mycena galopoda (Bühler, 

Kollar, 1996). Their biochemical action – inhibition of breath Qo; therefore, to the 

center in cytochrome b of mitochondria the abbreviated name QoI (Qo – inhibitors) is 

given. To the Site-specific fungicides high risk of resistance development is characteristic. 

In 1998 (Erichsen, 1999) for the first time it was informed on field resistance of 

downy mildew of wheat to strobilurine in the North of Germany. In 2001 there were 

data on resistance of cucumber powdery and downy mildew agents (Podosphaera 

fusca and Pseudoperonospora cubensis) in greenhouses of Japan (Ishii et al., 2001). 

A representative of QoI-fungicides krezoksim methyl has started to be used in Europe 

in industrial orchards since 1996 (Kunz et al., 1998). In America the application of 

this fungicide in industrial orchards began in 1999, and in the experimental ones – in 

1994. Since 1997 the researches on studying the mechanism of resistance occurrence 

and monitoring of sensitivity to strobilurine have been carried out and in 2000 a 

laboratory mutant of V. inaequalis resistant to krezoxym methyl is obtained (Olaya, 

Koller, 1999; Zheng, Olaya, Koller, 2000). The pathogen resistance is noted in North 

America (USA, Canada) and South America (Chile) (Koller et al., 2004; Sallato, 

Latorre, 2006; Jobin, Carisse, 2007). 

In this connection the objective of our work was to carry out V. inaequalis resistance 

to strobilurin group strobi fungicide (a. i. krezoxym methyl) monitoring in 

industrial orchards. 

Object, methods and conditions. The infected material (scab infected leaves 

and fruits) was selected from industrial orchards “Kletsky”, Minsk area, and “Rassvet”, 

Brest area, and also from private sector gardens, where there were no fungicide 

treatments. The monosporous isolates of V. inaequalis were isolated by means of a 

special nozzle and a cultivation method (Седов, Жданов, 1985). 

The pathogen sensitivity to krеzоxym methyl was estimated by the general in 

phytotoxicological practice method of fungus sowing on a nutrient medium (PDA), 

containing various krezoxym methyl concentrations (BАSF Co., 500 g/kg krezoxym 

methyl) (Koller et al., 2004). There were 4 repetitions, control – without fungicide. 

krezoxym methyl concentration from 2.5 to 200 mkg ml -1 (ррm) was prepared 

by Strobi (BАSF Co., 500g/kg krezoxym methyl) dilution in water. On 6, 12, 15, 21, 

30-th day the fungus colonies were measured and the percent of growth suppression 

using Ebbot formula (1) and relative growth (2) were calculated. 

Т = (Dk - Df) : Dk∙100; (1) 

where T – growth suppression in comparison with the control, %; Dk – diameter 

of colony in the control (on agar with fungicide), mm; Df – diameter of colony in 

experience (on agar without fungicide), mm; 

RG = Df : Dk∙100; (2) 

where RG – relative growth of pathogen colonies, %; Df – diameter of colony on 

agar with fungicide, mm; Dk – diameter of colony on agar without fungicide, mm. 

ЕС 50 

(Fungicide concentration, 50 % necessary for effective suppression of isolates, 

forms composing a population of the studied strain) was defined on a slide-rule 

where on axis X there were the data on fungus colonies growth suppression (%), on 

axis Y – concentration of studied fungicide in mkg ml -1 (probit-analysis). Resistance 

94

factor (RF) was calculated considering the relation of ЕС 50 

tested isolates value to 

average ЕС 50 

of sensitive isolates. As sensitive “wild” isolates were used, isolated 

from the orchard, which was not under pesticide treatments. 

Sensitivity of apple scab agent conidia to krezoxym methyl was estimated by the 

results of their germination in a drop of fungicide solution (a hanging drop) in various 

concentrations. In the control аscospores and conidia were germinated in a drop of 

water. The results were considered in 24, 48 and 72 hours (Голышин, 1993). 

For statistic experimental results processing (В.Ф. Moисейченко, 1988) the 

average arithmetic and confidence intervals detection for 95 % probability level on 

PC using Excel 2007 was carried out. 

Results. In orchard of farm “Kletsky”, Minsk area, since 2000 in the system of 

apple-tree protection against scab krezoxym methyl has been applied at the rate of 

0.2 kg ha -1 . The number of treatments has made four during a season. The analysis 

of V. inaequalis sensitivity showed that at the end of vegetative period all fungus 

isolates were sensitive to krezoxym methyl. Thus, in 44.1 % isolates ЕС 50 

reached 

20 mkg ml -1 , in 38.2 % isolates ЕС 50 

value fluctuated from 21 to 40 mkg ml -1 , and in 

14.7 % isolates has not increased 60 mkg ml -1 (Table 1). Krezoxym methyl concentration 

applied at orchard sprayings has made 100 mkg ml -1 . 

Table 1. Distribution of V. inaequalis isolates isolated from the industrial orchards 

by sensitivity to krezoxym methyl 

1 lentelė. Iš pramoninių sodų išskirtų V. inaequalis izoliatų pasiskirstymas pagal jautrumą 

krezoksim metilui 

Years of 

researches 

Tyrimų 

metai 

Number 

of isolates 

Izoliatų 

skaičius 

Number of 

treatments 

Distribution of isolates by ЕС 50 

Izoliatų pasiskirstymas pagal ЕС 50 

(%) 

41–60 61–80 81–100 > 100 

mkg ml -1 mkg ml -1 mkg ml -1 mkg ml -1 mkg ml -1 

Apdorojimų 

skaičius 

0–20 

mkg ml -1 21–40 

“Kletsky”, Мinsk district (krezoxym methyl application since 2000) 

“Кletsky”, Мinsko rajonas (krezoksim metilas naudojamas nuo 2000 m.) 

2000 34 4 44.1 38.2 14.7 0 0 0 

2001 56 4 14.3 23.2 25.0 12.5 3.6 21.4 

2002 60 0 1.7 13.3 13.3 28.3 6.7 36.7 

“Rassvet”, Brest district (krezoxym methyl application since 1998) 

“Rassvet”, Bresto rajonas (krezoksim metilas naudojamas nuo 1998 m.) 

2005 49 4 0 0 10.2 16.3 14.3 59.2 

2006 58 1 0 1.7 6.9 10.3 22.4 58.6 

In 2001 in the same orchard also 4 krezoxym methyl reatments were done. The 

biological efficiency of the fungicide was high and reached 90.2 %. However, by 

V. inaequalis isolation from the infected leaves during harvesting it was established 

that in 21.4 % of isolates ЕС 50 

value was higher than krezoxym methyl concentration, 

applied in orchard, what allows us to state that these isolates were resistant. Giving 

up krezoxym methyl treatments in 2002 has increased the quantity of such isolates 

1.7 times and has made 36.7 %. 

Under the conditions of Brest area (industrial orchard of the farm “Rassvet”) 

95

in the system of orchard protection the krezoxym methyl has been applied two-four 

times during a season since 1998. The efficiency of a preparation was high enough – 

the disease development did not exceed 7 %. However, in orchard “Rassvet” in 2005 

after 4 times of this fungicide application its efficiency has essentially decreased: the 

disease development on leaves reached 21.3 %, on fruits – 43.6 %. It was a reason 

for estimation the sensitivity of V. inaequalis populations to krezoxym methyl. It was 

established that a share of resistant isolates, in which ЕС 50 

value exceeded 100 mkg ml -1 , 

has made 59.2 %. 

In 2006 when orchard treatments were reduced up to one, the situation has 

not changed and the proportion of resistant isolates also remained at the same level 

(58.6 %). 

Having generalized the data obtained during investigations, we established a considerable 

decrease of V. inaequalis sensitivity to the krezoxym methyl from strobilurine 

group. In orchard “Kletsky”, Minsk area, in the first year of Strobi application ЕС 50 

of the pathogen population has made 26.6 mkg ml -1 and the resistance factor has not 

increased 24.2 (Table 2). 

After 4-fold application of a preparation in 2001 the average ЕС 50 

value and the 

factor of V. inaequalis resistance has increased 2.6 times and has made 68.6 mkg ml -1 

and 62.4, accordingly. 

Таble 2. Sensitivity of V. inaequalis populations from the industrial orchards of 

Belarus to krezoxym methyl 

2 lentelė. Pramoninių Baltarusijos sodų V. inaequalis populiacijų jautrumas krezoksim 

metilui 

Year of 

investigations 

Tyrimų metai 

Number of treatments 

per season 

Apdorojimų 

skaičius per sezoną 

Number of 

isolates 

Izoliatų 

skaičius 

Average ЕС 50 

(mkg ml -1 ) 

ЕС 50 

vidurkis, 

mkg ml -1 

Factor of 

resistance 

Atsparumo 

faktorius 

“Кletsky”, Мinsk district (krezoxym methyl application since 2000) 

“Кletsky”, Мinsko rajonas (krezoksim metilas naudojamas nuo 2000 m.) 

2000 4 34 26.6 ± 2.1 24.2 

2001 4 56 68.6 ± 7.2 62.4 

2002 0 60 97.2 ± 7.4 88.4 

“Rassvet”, Brest district (krezoxym methyl application since 1998) 

“Rassvet”, Bresto rajonas (krezoksim metilas naudojamas nuo 1998 m.) 

2005 4 49 128.9 ± 9.1 117.2 

2006 1 58 121.8 ± 6.1 110.8 

Note. ЕС 50 

of sensitive isolates – 1.1 ± 0.3 mkg ml -1 

Pastaba. Jautrių izoliatų ЕС 50 

– 1.1 ± 0.3 mkg ml -1 

When Strobi application was stopped in 2002, the tendency to sensitivity decrease 

remained: the average ЕС 50 

value of fungus population reached 97.2 mkg ml -1 , and 

resistance factor made 88.4. 

In orchard “Rassvet”, Brest area, where krezoxym methyl was applied since 

1998, after four times fungicide application in 2005 the average ЕС 50 

for V. inaequalis 

96

eached 128.9 mkg ml -1 , and resistance factor – 117.2. When frequency of treatments 

decreased to one in 2006 the situation has not changed: ЕС 50 

of the pathogen population 

reached 121.8 mkg ml -1 , and resistance factor – 110.2, what testifies the progressing 

loss of V. inaequalis population sensitivity to krezoksim methyl. 

ЕС 50 

of Kletsk population conidia germination in a fungicide drop has made 

35 mkg ml -1 (ЕС 50 

of mycelium growth – 97.2 mkg ml -1 ). ЕС 50 

value of conidia germination 

isolated from orchard “Rassvet” in 2005 has made 118 mkg ml -1 , in 2006 – 

94 mkg ml -1 (ЕС 50 

of mycelium growth – 128.9 and 121.8 mkg ml -1 , accordingly). Thus, 

the obtained results confirmed the presence of forms resistant to krezoxym methyl in 

the analyzed populations. 

Since strobilurine action is directed basically on pathogen conidia germination 

suppression, we carried out trials studying conidia sensitivity to krezoxym methyl. 

At krezoxym methyl concentration 500 mkg ml -1 , that is, 5 times exceeding the 

used in a orchard, 1.5 % of conidia isolated from the infected apple-tree samples 

from orchard “Kletsky”, Minsk district, has germinated and 5.5–7.2 % from orchard 

“Rassvet”, Brest district (Fig.). 

At 100 mkg/ml concentration – 32.8 % of V. inaequalis conidia from orchard 

“Kletsky” and 47.9–54.3 % of the pathogen conidia from orchard “Rassvet” has 

germinated. 

Fig. Germination of V. inaequalis conidia in different krezoxym methyl 

concentrations (laboratory trial) 

Pav. V. inaequalis konidijų sudygimas skirtingose krezoksim metilo 

koncentracijose (laboratorinis bandymas) 

97

Discussion. On getting the messages on apple scab agent resistance to QoIfungicides, 

an intensive monitoring took place in Europe. The diverse levels of resistance 

were found in all countries of the European Union from zero and up to the 

highest level even within one orchard (Broniarek-Niemiec, Bielenin, Dyki, 2002). 

During our researches it was also determined, that in the first year of application 

krezoxym-methyl efficiency against apple scab was rather high and all pathogen 

isolates were susceptible to the given fungicide. However, the longer this preparation 

is used in orchards (even at the allowed 4-times spraying), the lower susceptibility of 

the fungi V. inaequalis to the fungus. 

The researches of W. Koller et al.(2004) showed that V. inaequalis develops 

2-stage resistance to strobilurine: quantitative and qualitative (Koller et al., 2004). The 

quantitative answer of the pathogen population is stipulated by quantity of the done 

treatments in the orchards by these fungicides, and qualitative – by mutation presence 

in the site G143A of mitochondrial V. inaequalis in cytochrome b, then a fungus get 

an alternative breath. 

Our obtained results proved the existence of a quantitative resistance (loss of sensitivity 

with the increase of treatments number) in the fungus V. inaequalis. However, 

it is necessary to continue the problem study and determine if resistance is qualitative 

to krezoxym methyl of apple scab causal agent. 

On the base of carried out monitoring it is not impossible to predict the development 

of apple scab causal agent resistance to krezoxym methyl, but it is only possible 

to receive the information about the geographical distribution of the stable strains 

depending on the number of carried out treatments by this preparation, population 

resistance level changes in years, using it for substantiation the strategy and tactics of 

different mechanism of action fungicides application. 

Conclusions. A considerable decrease of sensitivity in apple scab agent V. inaequalis 

populations to krezoxym methyl (fungicide Strobi) from strobilurine group 

is revealed. 

The average ЕС 50 

value of isolates isolated from the orchard where krezoxym 

methyl was applied 4 times within 2 years has made from 26.6 to 97.2 mkg ml -1 . In 

21.4–36.7 % of isolates ЕС 50 

increased the preparation concentration used in orchards 

(100 mkg ml -1 ). 

Germination of 1.5 % conidia is noted at fungicide concentration 5 times exceeding 

the one applied in production. 

In orchards where krezoxym methyl was applied in the same frequency rate during 

7 years, ЕС 50 

value increased to 121.8–128.9 mkg ml -1 , number of isolates with 

ЕС 5 

> 100 mkg ml -1 have made 58.6–59.2 %, and at concentration ЕС 50 

exceeding 

the one applied in production 5 times the amount of germinated conidia has made 

5.5–7.2 %, what testifies to the progressing loss of sensitivity of V. inaequalis population 

to krezoxym methyl. 

Gauta 2009 06 30 


98

References 

1. Broniarek-Niemiec A., Bielenin A., Dyki B. 2002. Efekt wyniszczajacego dzialania 

fungicydow strobilurynowych i difenkonazolu na grzybnie i zarodnikowanie 

Venturia inaequalis. Acta agrobotanica, 55(1): 49–58. 

2. Bühler B., Kollar A. 1996. Untersuchungen zur Strobilurinwirkung auf die 

zellwandabbauenden Enzyme des Apfelschorfpilzes Venturia inaequalis. 

Mitteilungen der Biologische Bundesanstalt fur Land – und Forstwirtschaft, 

321: 576. 

3. Erichsen E. 1999. Problems in mildew control in northern Germany. Getreide, 

1: 44–46. 

4. Ishii H., Fraaije B. A., Sugiyama T., Noguchi K., Nishimura K., Takeda T., 

Amano T., Hollomon D. W. 2001. Occurrence and molecular characterization 

of strobilurin resistance in cucumber powdery mildew and downy mildew. 

Phytopathology, 91: 1 166–1 171. 

5. Jobin T., Carisse O. Incidence of Myclobutanil- and Kresoxim-Methyl- 

Insensitive Isolates of Venturia inaequalis in Quebec Orchards. Plant Disease, 

91(10): 1 351–1 358. 

6. Koller W., Parker D. M., Turechek W. W., Avila-Adame C. 2004. A Two-Phase 

Resistance Response of Venturia inaequalis Populations to the QoI Fungicides 

Kresoxim-Methyl and Trifloxystrobin. Plant Disease, 88: 537–544. 

7. Kunz S., Lutz B., Desing H., Mendgen K. 1998. Assessment of sensitivities to 

anilinopyrimidine – and strobilurin-fungicides in populations of the apple scab 

fungus Venturia inaequalis. Journal of Phytopathology, 146: 231–238. 

8. Olaya G., Koller W. 1999. Baseline Sensitivities of Venturia inaequalis 

Populations to the Strobilurin Fungicide Kresoxim-methyl. Plant Disease, 83: 

274–278. 

9. Sallato B. V., Latorre B. A. 2006. First Report of Practical Resistance to QoI 

Fungicides in Venturia inaequalis (Apple Scab) in Chile. Plant Disease, 90: 

375. 

10. Zheng D., Olaya G., Koller W. 2000. Characterization of laboratory mutants of 

Venturia inaequalis resistant to the strobilurin-related fungicide kresoxim-methyl. 

Current Opinion in Genetics and Development, 35: 148–155. 

11. Голышин Н.М. 1993. Фунгициды. Москва: Колос. 

12. Моисейченко В.Ф. 1988. Методика опытного дела в плодоводстве и 

овощеводстве. Киев: Выща школа. 

13. Самусь В. А. 2007. Агробиологические основы интенсификации 

производства плодов яблони в республике Беларусь: автореф. дис. д-ра 

с.-х. наук. Горки. 

14. Седов Е.Н., Жданов В.В. 1985. Методика отбора устойчивых к парше сортов 

и сеянцев яблони на искусcтвенных инфекционных фонах. Москва. 

99


Venturia inaequalis populiacijų jautrumas krezoksym metilui 

V. Kamardzina 

Santrauka 

2000–2006 metais pramoniniuose soduose atliktų tyrimų metu buvo nustatytas reikšmingas 

obelų rauplių sukėlėjo Venturia inaequalis populiacijų jautrumo krezoksim metilui 

(Strobi) iš strobilurino grupės sumažėjimas. Sode “Kletsky” (Minsko rajonas) pirmaisiais 

šio fungicido naudojimo metais jautrių izoliatų kiekis sudarė 100 %, o atsparumo faktorius 

neviršijo 24,2. 2001 metais panaudojus preparatą 4 kartus, jautrių izoliatų proporcija 

sumažėjo iki 80,2 %, o populiacijos atsparumo faktorius padidėjo 2,6 karto. Kai 2002 metais 

Strobi naudojimas buvo nutrauktas, jautrumo mažėjimo tendencija išliko: jautrių 

izoliatų proporcija sumažėjo iki 63,3 %, o populiacijos atsparumo faktorius pasiekė 88,4. 

Sode “Rassvet” (Bresto rajonas), kuriame krezoksim metilas buvo pradėtas naudoti nuo 

1998 metų, 2005 metais atsparių izoliatų proporcija sudarė 59,2 %, o populiacijos atsparumo 

faktorius buvo 117,2. Kai 2006 metais apdorojimų skaičius sumažėjo iki vieno, situacija iš 

esmės nepasikeitė, o tai rodo mažėjantį V. inaequalis populiacijos jautrumą šiam fungicidui. 

Reikšminiai žodžiai: atsparumas, fungicidai, obelys, pramoniniai sodai, strobilurinas, 

veiksminga koncentracija (VK 50 

), Venturia inaequalis. 

100




Efficacy of herbicide Lentagran WP for control of 

annual dicotyledonous weeds in cabbage crop 

Danguolė Kavaliauskaitė 


Lithuania, e-mail d.kavaliauskaite@lsdi.lt 

In 2007–2008 at the Lithuanian Institute of Horticulture there were carried out the investigations 

of herbicide Lentagran WP (a. i. pyridate 45 %) efficiency in cabbage crop. 

The investigated herbicide effectively decreased weed number in cabbage crop. Annual 

dicotyledonous weeds were sensitive to herbicide Lentagran WP 0.5–2.0 l ha -1 . Total number 

of weeds 14 days after application of herbicide decreased by 51.6–82.3 %, number of annual 

dicotyledonous weeds decreased by 58.3–82.5 %, air dry weight of weeds decreased by 34.9– 

67.6 %. The number of annual dicotyledonous weeds in Lentagran WP 0.5–2.0 l ha -1 treatments 

was essentially lower to compare with untreated and also there was found essentially lower 

number of annual dicotyledonous weeds in Lentagran WP (a. i. pyridate 45 %) 2.0 l ha -1 treatment 

to compare with Butizan 400 2.0 l ha -1 treatment. Especially sensitive to Lentagran WP 

0.5–2.0 l ha -1 there was Galinsoga parviflora Cav. (80.4–100 %). Very sensitive to Lentagran 

WP 2.0 l ha -1 were Matricaria inodora L. (93.7 %), Stellaria media L. (87.3 %). 

Key words: cabbage, herbicides, Lentagran WP, pyridate, yield, weeds. 

Introduction. Broadleaf weeds continue to be significant problem in cabbage 

production fields. Until recently only two herbicides were registered for postemergence 

broadleaf weed control in transplanted cabbage. With the recent registration 

of Lentagran WP the possibility of developing postemergence weed management 

strategies for both direct-seeded and transplanted cabbages exists. Herbicide Lentagran 

WP now registered for use in onion, leek and cabbage in the Lithuania, controls 

or suppresses redroot pigweed (Amaranthus retroflexus L.), common lambsquarters 

(Chenopodium album L.), nightshade spp. (Solanum spp.) and small flowered galinsoga 

(Galinsoga parviflora Cav.). Lentagran WP is postemergence contact herbicide 

and has no residual soil activity. Its mode of action involves hydrolysis to 3-phenyl- 

4-hydroxy-6-chloropyridazine, which then inhibits photosystem II electron transport 

(Zohner, 1987). 

Effective weed management must also have good crop tolerance. Pyridate injury, 

which occurs as distinctive, blotchy chlorosis of treated leaves, has frequently been 

reported in cabbage (Bullen et al., 1993; Miller, Hopen, 1991; Orfanedes, Masiunas, 

1990; Wallace, Bellinder, 1992). Four-leaf cabbage tolerated pyridate when applied at 

101

1x and 2x rates (1.0 and 2.0 kg/ha, respectively) however, when applied at earlier growth 

stages, significantly injury has been reported phytotoxicity. Injuring is usually transitory 

with nonchlorotic leaves emerging subsequent to application. There have been no 

reported instances of yield reductions in cabbage where pyridate was applied. 

The aim of the study – to investigate the effect of herbicide Lentagran WP (a. i. 

pyridate 45 %) efficiency in cabbage crop. 


Institute of Horticulture in 2007–2008. Soil – sandy loam on light loam, calcaric 

epihypogleyic luvisol (IDg 8-k, / Calc(ar)i – Epihypogleyc Luvisolls – LVg-p w cc). 

Ploughing layer was of 22–25 cm in thickness, of little humus (1.58 %), neutral 

(pH KCL 

7.0). There was big amount of agile phosphorus (354 mg kg -1 of the soil), potassium 

(146 mg kg -1 of the soil) and calcium (4 500 mg kg -1 of the soil) in the soil, 

but small amount of nitrogen (in the layer of 0–40 cm – 56.6 kg ha -1 N-NH 4 

+ N-NO 3 

). 

Before planting 200 kg ha -1 N, 150 kg ha -1 P, 200 kg ha -1 K were poured. There was 

grown cabbage cultivar ‘Langedijker Dauer’. Investigations were carried out according 

to the mechanized technology of cabbage growing created at the Lithuanian Institute 

of Horticulture. Cabbage experimental area was 600 m 2 . Record plot – 5 × 5 = 25 m 2 . 

Experiment was carried out in 4 replications. 

Herbicides in cabbage crop were sprayed two weeks after planting according to 

the s c h e m e: 

1) Control (without herbicides, weeded); 

2) Butizan 400 (standard) 2.0 l ha -1 ; 

3) Lentagran WP 0.5 l ha -1 ; 



6) Lentagran WP 2.0 l ha -1 . 

Herbicides were sprayed with back sprayer. Water rate – 400 l ha -1 . Weeds were 

counted in four places of every plot, in areas of 0.25 m 2 , diagonally thought the plot, 

once after the month from the last spraying. After weed calculation, they were weeded 

once. The data of crop weediness and cabbage yield were calculated by dispersion 

analysis method. Before carrying out statistical analysis, the number of weeds was 

recounted according to the formula: y = (x + 1): x – real weed date; y – transformed 

weed date (Tarakanovas, Raudonius, 2003). 

Results. In 2007–2008 the average total number of weeds after application of 

herbicide Lentagran WP 0.5–2.0 l ha -1 decreased by 51.6–82.3 % to compare with 

untreated. The number of total weeds in Lentagran WP 0.5–2.0 l ha -1 treatments was 

found essentially lower to compare with untreated and there was found essentially lower 

total number of weeds in Lentagran WP 1.5–2.0 l ha -1 treatments to compare with 

Butizan 400 2.0 l ha -1 standard treatment. The efficacy of Lentagran WP 1.5–2.0 l ha -1 

was bigger in 13.3–18.0 % to compare with standard treatment. Total weed number 

was lower in 2007 than in 2008, but herbicide efficacy was similar in both years of 

investigation (Table 1). 

102

Table 1. Efficiency of herbicide Lentagran WP for control of total number of 

weed in cabbage 

1 lentelė. Herbicido Lentagran WP įtaka bendram piktžolių skaičiui kopūstų pasėlyje 

Treatments 

Variantai 

Babtai, 2007–2008 

Total number of weeds (pcs. m -2 ) 

Bendras piktžolių skaičius, vnt. m -2 

2007 2008 

2007–2008 average 

vidurkis 

36.5 79.0 57.7 

Untreated 

Nepurkšta 

Butizan 400 2.0 l ha -1 (standard 18.2* 23.0* 20.6* 

standartas) 

Lentagran WP 0.5 l ha -1 27.2* 28.7* 27.9* 

Lentagran WP 1.0 l ha -1 19.0* 23.7* 21.3* 

Lentagran WP 1.5 l ha -1 8.2** 17.7* 12.9** 

Lentagran WP 2.0 l ha -1 6.2** 14.2** 10.2** 

Note: * – essentially less than in the untreated treatment (LSD 05 

), 

** – essentially less than in the Butizan 400 2.0 l ha -1 (standard) treatment (LSD 05 

) 

Pastaba: * – iš esmės mažiau negu nepurkštame variante (R 05 

), ** – iš esmės mažiau negu herbicidu 

Butizan 400 2,0 l ha -1 nupurkštame standartiniame variante (R 05 

). 

Average number of annual dicotyledonous weeds in 2007–2008 in herbicide 

Lentagran WP 0.5–2.0 l ha -1 treatment decreased by 58.3–82.5 % to compare with 

untreated. There was found essentially lower number of annual dicotyledonous 

weeds in Lentagran WP 0.5–2.0 l ha -1 treatments to compare with untreated and 

also there was found essentially lower number of annual dicotyledonous weeds in 

Lentagran WP 1.5–2.0 l ha -1 treatments to compare with Butizan 400 2.0 l ha -1 standard 

treatment. The efficacy of Lentagran WP 1.5–2.0 l ha -1 in post emergence dicotyledonous 

weed management was bigger in 14.8–16.7 % to compare with standard treatment. 

Number of annual dicotyledonous weeds was lower in 2007 and efficacy of Lentagran 

WP 2.0 l ha -1 was bigger by 5 % in that year than in 2008 (Table 2). 

Total air-dry weight of weeds in Lentagran WP 0.5–2.0 l ha -1 treatments decreased 

by 34.9–67.6 % to compare with untreated in both year of investigation. There was 

found essentially lower air-dry weight of weeds in Lentagran WP 0.5–2.0 l ha -1 treatments 

to compare with untreated and there was found essentially lower air-dry weight 

of weeds in Lentagran WP 2.0 l ha -1 treatment to compare with Butizan 400 2.0 l ha -1 

treatment. Lentagran WP 2.0 l ha -1 decreased air-dry weight of weeds in 27.6 % 

(Table 3). 

103

Table 2. Efficiency of herbicide Lentagran WP for control of annual dicotyledonous 

weed number in cabbage 

2 lentelė. Herbicido Lentagran WP įtaka vienmečių dviskilčių piktžolių skaičiui kopūstų 

pasėlyje 

Treatments 

Variantai 

Babtai, 2007–2008 

Number of annual dicotyledonous weeds, (pcs. m -2 ) 

Vienmečių dviskilčių piktžolių skaičius, vnt. m -2 

2007 2008 

2007–2008 average 

vidurkis 

33.5 78.5 55.6 

Untreated 

Nepurkšta 

Butizan 400 2.0 l ha -1 

16.2* 22.7* 19.0* 

(standard 

standartas) 



Lentagran WP 1.5 l ha -1 6.2** 15.5* 10.8** 



), ** – essentially less than in 

the Butizan 400 2.0 l ha -1 (standard) treatment (LSD 05 

). 




). 

Table 3. Efficiency of herbicide Lentagran WP for control of weed air-dry weight 

in cabbage 

3 lentelė. Herbicido Lentagran WP įtaka piktžolių orasausei masei kopūstų pasėlyje 

Treatments 

Variantai 

Babtai, 2007–2008 

Total air-dry weight of weeds 

Orasausė piktžolių masė (g m -2 ) 

2007 2008 

2007–2008 average 

vidurkis 

81.2 105.0 96.7 

Untreated 

Nepurkšta 

Butizan 400 2.0 l ha -1 (standard 51.2* 63.5* 57.7* 

standartas) 






), ** – essentially less than in 

the Butizan 400 2.0 l ha -1 (standard) treatment (LSD 05 

). 




). 

104

The number of Chenopodium album L. decreased by 74.8 % in treatments with 

Lentagran WP 2.0 l ha -1 and decreased by 49.8–67.7 % in Lentagran WP 0.5–1.5 l ha -1 

treatment and – 44.7 % in Butizan 400 2.0 l ha -1 standard treatment during investigation 

year. The number of Galinsoga parviflora Cav. decreased by 80.4–100 % in treatments 

with Lentagran WP 0.5–2.0 l ha -1 and decreased by 68.2 % in Butizan 400 2.0 l ha -1 

treatment. The number of Capsella bursa-pastoris L. decreased by 66.0–73.1 % in 

treatments with Lentagran WP 1.5–2.0 l ha -1 and – by 27.7–46.4 in treatments with 

Lentagran WP 0.5–1.0 l ha -1 and Butizan 400 2.0 l ha -1 treatment. The number of 

Stellaria media L. decreased by 87.3 % in treatments with Lentagran WP 2.0 l ha -1 and 

decreased by 100 % in treatment with Butizan 400 2.0 l ha -1 treatments. The number 

of Matricaria inodora L. decreased in Lentagran WP 2.0 l ha -1 treatment by 93.7 %, 

in Lentagran WP 1.0 l ha -1 – by 79.7 % and in Butizan 400 2.0 l ha -1 treatment – by 

17.2 % during investigation year (Table 4). 

Table 4. Efficiency of Lentagran WP for reduction (%) of main dicotyledonous 

weed species in cabbage 

4 lentelė. Kai kurių vienmečių dviskilčių piktžolių rūšių skaičiaus sumažėjimas (%) 

panaudojus herbicidą Lentagran WP kopūstų pasėlyje 

Babtai, 2007–2008 

Treatments Chenopodium Galinsoga Stellaria Matricaria Capsella 

Variantai album L. parviflora Cav. media L. inodora L. bursa-pastoris L. 

Untreated 

0 0 0 0 0 

Nepurkšta 

Butizan 400 2.0 l ha -1 44.7 68.2 100 17.2 34.6 


Lentagran WP 0.5 l ha -1 49.8 96.5 64.7 14.0 27.7 



Lentagran WP 2.0 l ha -1 74.8 100 87.3 93.7 73.1 

There was not found significant differences between treated with herbicides 

Lentagran WP 0.5–2.0 l ha -1 , Butizan 400 2.0 l ha -1 treatments and untreated in cabbage 

yield in both year of investigation. Negative direct effect on crop, cabbage yield and 

exterior quality of yield was no observed during investigation year (Table 5). 

105

Table 5. Yield of cabbage after application of herbicide Lentagran WP 

5 lentelė. Kopūstų derlius panaudojus herbicidą Lentagran WP 

Treatments 

Variantai 

Babtai, 2007–2008 

Marketable yield 

Total yield 

Prekinis derlius 

Bendras derlius 

% from total yield 

(t ha -1 ) 

t ha -1 

% nuo bendro derliaus 

39.8 39.0 97.9 

Untreated 

Nepurkšta 

Butizan 400 2.0 l ha -1 (standard 43.1 42.8 99.3 

standartas) 

Lentagran WP 0.5 l ha -1 46.3 43.9 94.8 




LSD 05 

/ R 05 

6.81 6.47 - 

Discussion. According to the data by Kathleen et al. (1990), herbicide pyridate 

applied postemergence on direct-seeded broccoli. According to Henderson and Cairns 

(2002), pyridate kills weeds in broccoli, Chinese cabbage, cabbage, or cauliflower 

with minimal crop damage. Stall and Hensel (1994) reports about pyridate spraying 

in onion. Applying 450 g ha -1 pyridate caused chlorotic spotting of the sprayed vegetable 

leaves, but did not affect marketable yields of broccoli, cabbage or cauliflower. 

This rate controlled deadnettle (Lamium amplexicaule), reduced sowthistle (Sonchus 

oleraceus) growth by only 30–50 % compared with an unweeded control. Cabbage 

and cauliflower yields were unaffected by spraying 900 g ha -1 pyridate. This rate 

improved sowthistle control to a commercially acceptable level (Henderson, Cairns, 

2002). In our investigation Lentagran WP (a. i. pyridate 45%) 1.0–1.5 l ha -1 showed 

the biggest efficient in cabbage. The number of annual dicotyledonous weeds decreased 

by 58.3–82.5 %. The number of Chenopodium album L. decreased by 74.8 % 

after spraying of Lentagran WP 2.0 l ha -1 . The number of Galinsoga parviflora Cav. 

decreased by 80.4–100 % after Lentagran WP 0.5–2.0 l ha -1 . The number of Capsella 

bursa-pastoris L. decreased by 66.0–73.1 % after Lentagran WP 1.5–2.0 l ha -1 . The 

number of Stellaria media L. decreased by 87.3 % after Lentagran WP 2.0 l ha -1 . The 

number of Matricaria inodora L. decreased in Lentagran WP 2.0 l ha -1 treatment by 

93.7 %, in Lentagran WP 1.0 l ha -1 – by 79.7 %. Negative effect of Lentagran WP 

1.0–1.5 l ha -1 on crop and cabbage yield was no observed. Orfanedes and Masiunas 

(1990) also reported, that four leaf cabbage tolerate pyridate when applied 1.0 and 

2.0 kg/ha. Bellinder et al. (1997) reported about sethoxydim and crop oil concentrate 

increase pyridate phytotoxycity in transplanted cabbage. 

106

Conclusions. 1. Lentagran WP (a. i. pyridate 45 %) 1.0–1.5 l ha -1 was most effective 

to control small dicotyledonous weeds in cabbage. 

2. Negative effect of Lentagran WP (a. i. pyridate 45 %) 1.0–1.5 l ha -1 on crop, 

cabbage yield and exterior quality was no observed. 

Gauta 2009 07 29 


References. 

1. Bellinder R. R., Kirkwyland J., Wallase R. W., Arsenovic M. 1997. Sethoxydim 

and Crop Oil Concentrate Increase Pyridate Phytotoxicity in Transplanted 

Cabbage (Brassica oleracea). Weed Technology, 11: 81–87. 

2. Bullen M. R., Cornes D. W., Ryan P. J. 1993. The crop tolerance of cabbage, 

Brussels sprouts and onions to pyridate. Brighton Crop Protection Conference, 

3:1 047–1 052. 

3. Henderson C. W., Cairns R. 2002. Post emergence spraying of clopyralid, picloram 

or pyridate in broccoli, Chinese cabbage, cabbage, or cauliflower kills weeds, 

with minimal crop damage. Australian Journal of Experimental Agriculture, 

42(8): 1 113–1 117. 

4. Kathleen A., Herbst A., Derr J. F. 1990. Effect of Oxyfluorfen, Pyridate, and 

BAS 514 Aplied Postemergence on Direct-seeded Broccoli (Brassica oleracea 

var. botrytis). Weed Technology, 4(1): 71–75. 

5. Miller A. B., Hopen H. J. 1991. Critical weed control period in seeded cabbage 

(Brassica oleracea var. capitata). Weed Technology, 5: 852–857. 

6. Orfanedes S. M., Masiunas J. B. 1990. Herbicide evaluation in trans-planted 

cabbage. Proc. North Central Weed Science Society, 47: 27–33. 

7. Stall W. M., Hensel D. R. 1994. Onion herbicide evaluation in North Florida. 

Proc. Florida State Horticulture Society, 107: 153–155. 


analizė taikant kompiuterines programas Anova, Stat, Split-plot iš paketo 

Selekcija ir Irristat. Akademija, Kėdainių r. 

9. Wallace R. W., Bellinder R. R. 1992. Alternative tillage and herbicide option for 

successful weed control in vegetables. HortScience., 27: 745–749. 

10. Zohner A. 1987. Mode of crop tolerance to pyridate in corn and peanuts. 1987. 

Br. Crop Protection Conference. Weed, 3: 1 083–1 090. 

107


Herbicido Lentagran WP veiksmingumas kopūstų pasėlyje 

naikinant vienametes dviskiltes piktžoles 

D. Kavaliauskaitė 

Santrauka 

2007–2008 m. Lietuvos sodininkystės ir daržininkystės institute buvo atlikti herbicido 

Lentagran WP (v. m. pyridate 45 %) veiksmingumo tyrimai kopūstų pasėlyje. 

Tirtas herbicidas veiksmingai sumažino bendrą piktžolių skaičių kopūstų pasėlyje. 

Vienametės dviskiltės piktžolės buvo jautrios herbicidui Lentagran WP 0,5–2,0 l ha -1 . 

Bendras piktžolių skaičius po herbicido purškimo praėjus 14 dienų sumažėjo 51,6–82,3 %, 

vienamečių dviskilčių piktžolių skaičius – 58,3–82,5 %, orasausė piktžolių masė – 

34,9–67,6 %. Vienmečių dviskilčių piktžolių skaičius buvo iš esmės mažesnis nupurškus 

Lentagran WP 0,5–2,0 l ha -1 negu nepurkštame variante, o Lentagran WP 2,0 l ha -1 – iš esmės 

mažesnis negu herbicidu Butizan 400 2,0 l ha -1 nupurkštame standartiniame variante. 

Herbicidui Lentagran WP 0,5–2,0 l ha -1 ypač jautri buvo smulkiažiedė galinsoga 

(Galinsoga praviflora Cav.) (80,4–100 %). Lentagran WP 2,0 l ha -1 labai jautrūs buvo bekvapis 

šunramunis (Matricaria inodora L.) (93,7 %) ir daržinė žliūgė (Stellaria media L.) (87,3 %). 

Reikšminiai žodžiai: derlius, herbicidai, kopūstai, Lentagran WP, piktžolės, pyridate. 

108




Tolerance of apple propagation material to herbicides 

Darius Kviklys 


Lithuania, e-mail d.kviklys@lsdi.lt 

Investigations were conducted in the commercial nursery of the Lithuanian Institute of 

Horticulture. In 2001–2003 herbicides Stomp (pendimethalin, 4 l ha -1 ), Goltix (metamitron, 

3.0 l ha -1 ), Lontrel 300 (0.3 l ha -1 ), Agil (propikvizafop, 1.5 l ha -1 ), Focus Ultra (cycloxydim, 

4.0 l ha -1 ), Fuzilade Super (fluazifop, 3.0 l ha -1 ) and combination Fuzilade Super (2.0 l ha -1 ) 

and Betanal Progress (phenmedipham, desmedipham and ethofumesate, 2.0 l ha -1 ) were 

tested in apple nursery during the first and second growing season. Herbicides were sprayed 

directly on plants without mechanical protection. Herbicides Stomp (4 l ha -1 ), Agil (1.5 l ha -1 ), 

Focus Ultra (4.0 l ha -1 ) and Betanal Progress (2.0 l ha -1 ) are safe to use in apple tree nursery. 

Herbicide Goltix (3 l ha -1 ) should be used during the second year of apple growth. Fusilade 

Super (3.0 l ha -1 ) and Lontrel 300 (0.3 l ha -1 ) caused leaf damages of one-year-old apple trees, 

but did not interfere to the final growth. Interaction between herbicides and cultivars were 

noticed in the experiment with one-year-old plants. 

Key words: apple propagation material, herbicide, vegetative growth, weed control. 

Introduction. Weed control in fruit tree nurseries is performed by mechanical 

means and herbicides. Application of soil herbicides have been the most common 

practice for many years, though many of used soil herbicides were hazardous 

pollutant. Nevertheless until recent years such herbicides like simazin, devrinol are 

still used in commercial nurseries legally in some countries or not legally in others 

(Strandberg, Scott-Fordsmann, 2002; Kopytowski et al., 1999; Wycior et al., 1999). 

Use of methyl bromide was wide spread soil preparation practice both for soil disinfection 

and weed control, but it is forbidden because of environmental concerns 

(Shrestha et al., 2008; Duniway, 2002). New knowledge on chemical degradation of 

chemicals in soil and water is gained and some of previously safe substances appear 

to be more aggressive than it is declared (Saratovskikh et al., 2007). 

Application of non-selective or broad-spectrum herbicides in the nursery requires 

special technique and sometimes it is risky since these herbicides can negatively affect 

or kill nursery plants. Growing bud and trees during the first season in the nursery are 

especially sensitive to herbicides. 

109

Use of more nature friendly selective herbicides is wide spread practice in growing 

of different agriculture crops (Kavaliauskaitė et al., 2008). Application of them in fruit 

tree nursery is very limited. Therefore, larger experiments were planned aiming on 

suitability to use various herbicides in apple tree nursery. 

Object, methods and conditions. Investigations were conducted in the commercial 

nursery of the Lithuanian Institute of Horticulture. In 2001–2002 following 

herbicides were tested on one-year-old apple propagation material (budded in summer 

2000): Stomp (active substance pendimethalin at the rate 4.0 l ha -1 ), Goltix (active 

substance metamitron, 3.0 l ha -1 ), Lontrel 300 (active substance clopyralid, 0.3 l ha -1 ), 

Agil (active substance propikvizafop, 1.5 l ha -1 ), Focus Ultra (active substance cycloxydim, 

4.0 l ha -1 ), Fuzilade Super (active substance fluazifop, 3.0 l ha -1 ) and combination 

Fuzilade Super (2.0 l ha -1 ) and Betanal Progress (active substances phenmedipham, 

desmedipham and ethofumesate, 2.0 l ha -1 ). In 2002–2003 the same herbicides 

were tested on two-year-old apples. Manual weeding was control treatment. Two 

apple cultivars ‘Auksis’ and ‘Shampion’ on B.396 rootstock were tested. 

Herbicides were applied two times during the early season: in the middle of May 

and at the end of June. Herbicides were sprayed by manual sprayer directly on plants 

without mechanical protection. Manual weeding was performed at the same time as 

application of herbicides and additionally in July. 

Trial consisted of four replications with 15–20 plants in each. 

Plant height (cm) was measured in May before application of herbicides, at the 

beginning of July and at the end of vegetation season. Trunk diameter (mm) was measured 

at the end of experiment. Stem and leaf damages were monitored and described 

in two weeks after application of herbicides. 

Growth characteristics of the planting materials were different and depended on 

climatic conditions of the vegetative period and cultivar. Since there were no interactions 

between year and used herbicides in the experiment with one-year-old apple 

planting material, the results are presented as an average of two years. No interactions 

were recorded between year, herbicides and cultivar in the experiment with two-yearold 

apple planting material; therefore, the results are presented as an average of two 

years and two cultivars. 

Variance analysis of tree growth characters was done using Fischer least significant 

difference test at P < 0.05. 

Results. No one of applied herbicides reduced growth of one-year-old apple material 

of cv. ‘Auksis’ (Table 1). Neither tree growth dynamics, nor final stem diameter 

significantly differed between herbicide treated and control plants. 

Herbicides Goltix, Fusilade Super and Lontrel 300 had negative effect on the 

height of cv. ‘Shampion’ during the earliest application date (Table 2). More pronounced 

tendencies of reduced tree height and stem diameter remained in plots sprayed 

by Goltix. 

110

Table 1. Herbicide effect on vegetative growth dynamics of one-year-old 

‘Auksis’ planting material 

1 lentelė. Herbicidų įtaka vienerių metų dauginamosios medžiagos ‘Auksis’ vegetatyvinio 

augimo dinamikai 

Herbicide 

Herbicidas 

Height 

Aukštis (cm) 

Stem diameter 

Kamieno skersmuo (mm) 

06.01 07.15 09.20 09.20 

39.8 85.6 139.0 13.2 

Control 

Kontrolė 

Goltix 41.0 82,1 137.2 12.7 

Stomp 38.9 84.2 139.7 13.0 

Fusilade Super 37.7 83.2 138.9 13.2 

Agil 41.0 86.5 142.3 13.3 

Focus Ultra 38.9 86.0 141.5 13.4 

Fuzilade Super + Betanal Progres AM 39.7 85.4 141.6 13.4 

Lontrel 300 38.4 84.2 143.0 13.7 

LSD 05 

/ R 05 

3.04 4.31 5.28 0.62 

Table 2. Herbicide effect on vegetative growth dynamics of one-year-old 

‘Shampion’ planting material 

2 lentelė. Herbicidų įtaka vienerių metų dauginamosios medžiagos ‘Shampion’ vegetatyvinio 

augimo dinamikai 

Herbicide 

Herbicidas 

Height 

Aukštis (cm) 

Stem diameter 


06.01 07.15 09.20 09.20 

34.5 70.7 128.7 12.7 

Control 

Kontrolė 

Goltix 30.2 65.9 125.2 12.2 

Stomp 33.2 68.9 127.8 12.6 


Agil 34.2 71.2 128.2 12.5 

Focus Ultra 34.8 70.5 129.3 12.9 

Fuzilade Super + Betanal Progres AM 33.2 69.2 128.1 12.3 

Lontrel 300 30.4 67.2 129.7 12.8 

LSD 05 

/ R 05 

3.05 4.92 6.2 0.61 

No one of used herbicides had negative effect on the vegetative characteristics 

of two-year-old apple planting material (Table 3). Only tendency of slightly reduced 

tree height was noticed in Goltix treatment. 

Cv. ‘Shampion’ was somewhat more sensitive than cv. ‘Auksis’. Lontrel 300 

caused leaf damages during both applications. Fuzilade Super damaged leaves of 

cv. ‘Auksis’ during first application, and leaves of cv. ‘Shampion’ during early and 

late applications. Herbicide Goltix was a reason of leaf chlorosis of cv. ‘Shampion’. 

Herbicides Agil, Focus Ultra and Stomp did not damaged leaves of any cultivar during 

both times of application. No leaf damages were recorded on 2-year-old apple 

propagation material. 

111

Table 3. Herbicide effect on vegetative growth dynamics of two-year-old apple 

planting material (average of two cultivars and two years) 

3 lentelė. Herbicidų įtaka dvejų metų dauginamosios medžiagos vegetatyvinio augimo 

dinamikai (dviejų veislių ir dvejų metų vidurkiai) 

Herbicide 

Herbicidas 

Height 

Aukštis (cm) 

Stem diameter 


06.01 07.15 09.20 09.20 

116.2 145.3 188.3 18.1 

Control 

Kontrolė 

Goltix 112.3 140.6 180.0 18.3 

Stomp 114.2 142.4 184.2 17.9 


Agil 115.4 144.9 187.4 18.2 

Focus Ultra 115.9 144.2 187.1 17.7 

Fuzilade Super + Betanal Progres AM 115.6 144.8 187.3 17.9 

Lontrel 300 116.2 145.1 189.8 18.0 

LSD 05 

/ R 05 

4.01 5.33 9.19 1.13 

Leaf damages by some herbicides were recorded on one-year-old planting material 

(Table 4). 

Table 4. Herbicide caused damages on apple propagation material 

4 lentelė. Herbicidų žala obelų dauginamajai medžiagai 

Herbicide 

Herbicidas 

1-year-old cv. ‘Auksis’ 

‘Auksis’ vienmečiai 

sodinukai 

1-year-old cv. ‘Shampion’ 

‘Šampion’ vienmečiai 

sodinukai 

2-year-old apple propagation 

material 

Obelų dvimečiai sodinukai 

06.01 07.15 06.01 07.15 06.01 07.15 

1 2 3 4 5 6 7 

Control - * - - - - - 

Kontrolė 

Goltix - - light chlorosis 

- - - 

lengva chlorozė 

Stomp - - - - - - 

Fusilade 

Super 

scorched 1 

to 3 leaves 

nudeginti 

1–3 lapai 

- scorched 1 

to 5 leaves 

nudeginti 

1–5 lapai 

scorched 1 

to 2 leaves 

nudeginti 

1–2 lapai 

- - 

Agil - - - - - - 

Focus Ultra - - - - - - 

Fuzilade Super 

+ Betanal 

Progress AM 

- - scorched 1 

to 3 leaves 

nudeginti 

1–3 lapai 

- - - 

112

1 2 3 4 5 6 7 

Lontrel 300 scorched 1 scorched 1 scorched 1 scorched 1 - - 

to 3 leaves 

nudeginti 

1–3 lapai 

to 2 leaves 

nudeginti 

1–2 lapai 

to 5 leaves 

nudeginti 

1–5 lapai 

to 3 leaves 

nudeginti 

1–3 lapai 

* no damages recorded / pažeidimų nenustatyta 

Discussions. Weed control in fruit tree nurseries continues to be a major problem. 

Weeds can decrease nursery plant development, interfere with field and harvest operations. 

Still extensive hand labour and tillage are used for weed control during the 

growing season. Since the costs of both fuel and labour continue to rise, herbicides are 

likely to become a more important weed management tool in the tree nursery industry 

(Hanson, Schneider, 2008). 

Herbicides availability is different throughout the world. European Commission 

continuously revises list of substances allowed to use for crop protection and many 

of them are banned or suggested to remove from the list in coming years. In some 

countries still widely used simazin appears to be phytotoxic to many species even at 

rates below recommended (Strandberg, Scott-Fordsmann, 2002). Proper replacement 

of simazin is an urgent task for fruit and nurseries growers. In our trial two soil herbicides 

Goltix and Stomp were tested. Both of them protect weed infestation for shorter 

time compared with simazin (Rankova et al., 2009 a). Negative apple tree reaction to 

Goltix was noticed immediately after its application. Tendency of reduced tree height 

was established both in one-year-old and two-year-old plantings. 

Another soil herbicide Stomp was not toxic for apple propagation material, what 

is confirmed in other trials too (Gercheva et al., 2002, Rankova et al., 2009 b), though 

there is data on its negative effect on apple stem diameter (Wycior et al., 1999). 

Herbicides Agil, Focus and Fusilade Super are used to control graminaceous weeds. 

Nevertheless, at early stage of plant development higher dose of Fusilade Super (3.0 

l ha -1 ) caused damages when it was applied directly on growing one-year-old shoots. 

Other herbicides Agil and Focus were safe to use at any plant development stage. 

Lontrel 300, which is used for control of a wide range of broadleaf weeds, damaged 

young apple leaves. In spite of visual damages it did not course any growth 

suppression for cv. ‘Auksis’ and final characteristics of one-year-old plants of both 

tested cultivars. 

Interactions between herbicides and cultivars were noticed in the experiment with 

one-year-old plants during the earliest application date. Vegetative growth of apple 

trees of cv. ‘Shampion’ is less luxuriant than cv. ‘Auksis’ and it could be a reason of 

its somewhat higher sensitivity to some herbicides. If there were no significant differences 

between herbicide treatments and not sprayed plots for cv. ‘Auksis’, some 

herbicides as Goltix, Fusilade Super and Lontrel 300 reduced significantly the height 

of cv. ‘Shampion’ during the first application. 

Conclusions. Preemergence herbicide Stomp (4.0 l ha -1 ) is safe to control weeds 

in apple nursery during the first and second year of plant growth. Herbicide Goltix 

(3.0 l ha -1 ) should be used during the second year of apple growth. 

Fusilade Super (3.0 l ha -1 ) and Lontrel 300 (0.3 l ha -1 ) damaged leaves of one- 

113

year-old apple trees when they were applied directly. This did not interfere to the final 

characteristics of propagation material. 

Herbicides Agil (1.5 l ha -1 ), Focus Ultra (4.0 l ha -1 ) and Betanal Progress (2.0 l ha -1 ) 

are safe to use in apple tree nursery. 

Tolerance to herbicides is determined by apple cultivar. 

Gauta 2009 06 30 


References 

1. Duniway J. M. 2002. Status of chemical alternatives to methyl bromide for preplant 

fumigation of soil. Phytopathology. 92: 1 337–1 343. 

2. Gercheva P., Rankova Z., Ivanova K. 2002. In vitro test system for herbicide 

phytotoxicity on mature embryos of fruit species. Acta Horticulturae (ISHS). 

577: 333–336. 

3. Hanson B. D., Schneider S. A. 2008. Evaluation of weed control and crop safety 

with herbicides in open field tree nurseries. Weed Technology, 22(3): 493–498. 

4. Kavaliauskaitė D., Dambrauskienė E., Zalatorius V., Viškelis P. 2008. Influence 

of herbicides on the productivity and raw material quality of the first year medicinal 

thyme. Sodininkystė ir daržininkystė, 27(4): 253–260. 

5. Kopytowski J., Jastrzebska M., Banaszkiewicz T. 1999. Effect of herbicides on 

primary and secondary weed infestation of fruit tree nursery. Acta Academiae 

Agriculturae ac Technicae Olstenensis. 3: 85–97. 

6. Rankova Z., Koumanov K. S., Kolev K., Shilev S. 2009 a. Herbigation in a 

cherry orchard – efficiency of pendimethalin. Acta Horticulturae (ISHS). 825: 

459–464. 

7. Rankova Z., Nacheva L., Gercheva P. 2009 b. Growth habits of the vegetative 

apple rootstock MM106 after treatment with some soil herbicides under in vitro 

conditions. Acta Horticulturae (ISHS). 825: 49–54. 

8. Saratovskikh E. A., Polyakova O. V., Roshchupkina O. S., Lebedev A. T. 2007. 

Products of the Photolysis of 3,6-Dichloropicolinic Acid (the Herbicide Lontrel) 

in Aqueous Solutions. Applied Biochemistry and Microbiology. 43(2): 227– 

231. 

9. Shrestha A. , Browne G. T., Lampinen B. D. , Schneider S. , Simon L. , Trout 

T. 2008. Perennial crop nurseries treated with methyl bromide and alternative 

fumigants: Effects on weed seed viability, weed densities, and time required for 

hand weeding. Weed Technology. 22: 267–274. 

10. Strandberg M. T., Scott-Fordsmand J. J. 2002. Field effects of simazine at lower 

trophic levels – a review. The Science of the Total Environment. 296(1–3): 

117–137. 

114

11. Wycior S., Kiczorowski P., Wojcik I. 1999. Influence of herbicides on growth of 

the apple trees cv. Red Elstar ‘Elshof’ in nursery. Annales-Universitatis-Mariae- 

Curie-Sklodowska. Sectio-EEE, Horticultura. 7: 7–13. 


Obelų dauginamosios medžiagos tolerantiškumas herbicidams 

D. Kviklys 

Santrauka 

2001–2003 m. Lietuvos sodininkystės ir daržininkystės instituto medelyne atlikti herbicidų 

stomp (pendimethalin, 4,0 l ha -1 ), goltix (metamitron, 3,0 l ha -1 ), lontrel 300 (0,3 l ha -1 ), agil 

(propikvizafop, 1,5 l ha -1 ), focus ultra (cycloxydim, 4,0 l ha -1 ), fuzilade super (fluazifop, 3,0 l ha -1 ) 

ir herbicidų derinio fuzilade super (2,0 l ha -1 ) ir betanal progress (phenmedipham, desmedipham 

ir ethofumesate, 2,0 l ha -1 ) tyrimai su vienmečiais ir dvimečiais obelų sodinukais. Visi herbicidai 

buvo purškiami nenaudojant apsaugos. Herbicidai stomp, agil, focus ir betanal progress tinkami 

naudoti obelų medelyne. Herbicidą goltix rekomanduojama naudoti dauginant dvimetes obelis. 

Didesnės normos fusilade super (3,0 l ha -1 ) ir lontrel apdegina vienmečių obelaičių lapus, tačiau 

neturi neigiamos įtakos galutiniam obelaičių dydžiui. Pirmamečiame medelyne nustatytos obelų 

veislės ir herbicidų sąveika. 

Reikšminiai žodžiai: herbicidai, kova su piktžolėmis, obelų dauginamoji medžiaga, 

vegetatyvinis augimas. 

115




Influence of maturity stage on fruit quality 

during storage of ‘Shampion’ apples 

Nomeda Kviklienė, Alma Valiuškaitė 


Lithuania, e-mail n.kvikliene@lsdi.lt 

Influence of fruit maturity on apple cv. ‘Shampion’ storage ability and rot development 

was investigated at the Lithuanian Institute of Horticulture in 2004–2005. Fruits for storage 

were harvested 5 times at weekly intervals before, during and after predictable optimum 

harvest date. Quality changes, presence of storage disorders, mass losses were measured 

during harvest period and at the end of storage. During investigation period fruit quality 

parameters changed according to harvest date and were specific for each trial year. Later 

harvested fruits were softer and had higher content of soluble solids. Fruit storage ability was 

closely connected to fruit maturity too. Apples were of the best quality at the end of storage, 

when maturity index at picking date was 0.22–0.17. During storage ‘Shampion’ apple rot was 

caused by Monilinia sp., Gloeosporium spp. and Penicillium spp. On the average apple fruits 

were mostly infected by fungus of Gloeosporium genus. 

Key words: Malus × domestica, maturity index, rots, storage, weight loss. 

Introduction. Different picking date of apple fruits during the harvest season 

may have a significant impact on fruit quality (Vielma et al., 2008; Rizzolo et al., 

2006; Kvikliene, 2001; Franelli, Casera, 1996; Streif, 1996). To ensure the highest 

fruit quality at the end of long storage, apples must be harvested mature but not fully 

ripe. If harvested too early fruits are smaller, have reduced flavour and colour, and 

are more susceptible to scald, bitter rot and internal breakdown. Mass reduction by 

water loss is greater in earlier picked apples because waxy surface is not completely 

formed at this moment (Zerbini et al., 1999; Juan et al., 1999). Early picked fruits 

are smaller and their surface in a storage unit is larger. Because water transpiration 

depends on fruit surface area too, small fruits loss their weight faster. Another reason 

of more intensive evaporation is structure of fruit cuticle, which is not fully developed 

when fruits are harvested too early. At the same time the cuticle is the first barrier 

that pathogens have to challenge (Ihabi et al., 1998). Later picked apples often are 

over-mature and all physiological processes, which complicate storage, even under 

optimal conditions, are underway (Ingle et al., 2000; Braun et al., 1995). Apples 

harvested too late are vulnerable to mechanical injures, sensitive to low temperature 

breakdown, water core and more rot (Hribar et al., 1996). At optimal harvest time 

picked apples have the organoleptic qualities (Casals et al., 2006), which enable them 

117

to survive more than six months of storage. 

The objective of this study was to investigate the effect of harvest time on fruit 

quality and storage ability of cv. ‘Shampion’ apples. 

Object, methods and conditions. Investigations were carried out with apple cv. 

‘Shampion’ on M.9 rootstock in 2004 and 2005. The measurements of fruit quality 

changes were performed 2–3 weeks before and after the predictable optimum harvest 

date. Apples for long storage were harvested 5 times at weekly intervals. The experiment 

was carried out with 4 replications and 20 trees per plot. 

On each picking date 10 fruits from each replication were taken for laboratory 

measurements: fruit firmness (kg cm -2 , measured with penetrometer FT-327 with 11 mm 

diameter probe), soluble solids content (%, with refractometer), starch conversion (with 

0.1 N iodine and potassium iodine solution, according to the scale 1–10). Maturity 

index was calculated as F/RS, where F – firmness, R – soluble solids concentration, 

S – starch conversion. 

On each picking date 100 fruits from each replication were taken in order to measure 

storability (firmness, soluble solids concentration, weight loss, storage disorders 

and rots). Fruits were stored for 180 days. 

The incidence (%) of fruit rot was established according to the f o r m u l a: 

A = B / C · 100 %; A – incidence of fruit rot; B – the number of samples, in which the 

rot has been detected, C – total number of investigated samples. 

Variance analysis of the main quality characters was done using ‘ANOVA’ statistical 

program. 

Results. During ripening period fruit firmness decreased by 13–15 % (Table 1). 

In both trial years significant differences were recorded starting from the 3 rd harvest. 

During the last week of investigation fruit softening was not significant. 

Table 1. Effect of harvest time on fruit quality during ripening 

1 lentelė. Skynimo laiko įtaka obuolių kokybei skynimo metu 

Harvest 

Skynimai 

Firmness 

Kietumas (kg cm -2 ) 

Soluble solids content 

Tirpios sausosios 

medžiagos (%) 

average 

2004 2005 

vidutiniškai 

Maturity index 

Sunokimo indeksas 

2004 2005 

average 

vidutiniškai 

2004 2005 

1 7.9 8.0 8.0 10.6 10.8 10.7 0.33 0.54 0.44 

2 7.8 7.9 7.9 10.7 11.2 11.0 0.22 0.36 0.29 

3 7.3 7.4 7.4 11.1 11.9 11.5 0.14 0.17 0.16 

4 6.8 6.9 6.9 10.9 11.2 11.1 0.09 0.11 0.10 

5 6.9 6.8 6.9 10.6 11.7 11.2 0.08 0.09 0.09 

LSD 05 

/ R 05 

0.24 0.37 0.29 0.25 0.20 0.27 0.014 0.023 0.021 

average 

vidutiniškai 

The dynamic of soluble solids content (SSC) was similar each year. The highest 

amount of SSC was observed in the third week of measurements, after which it levelled 

off. 

Maturity index decreased linearly to harvest date. In 2005 at 1st harvest the value 

of maturity index was much higher. In spite of that, fruit maturation process was much 

faster and no significant differences were found at last harvest among year. 

118

At the end of storage fruit firmness gradually decreased according to harvest time 

and in most cases differences between dates were significant (Table 2). During storage, 

fruit firmness decreased on the average from 47 to 54 % of its original value. 

During the 180 days of storage period SSC on the average increased by 6–15 % 

from its original value at harvest. In 2004 the highest SSC was found at 1st harvest 

after which it levelled off, whereas in 2005 value of SSC linearly depended on harvest 

time, and the highest amount was observed in fruits of the two last harvests. 

Table 2. Effect of harvest time on fruit quality at the end of storage 

2 lentelė. Skynimo laiko įtaka obuolių kokybei laikymo pabaigoje 

Harvest 

Firmness 

Kietumas (kg cm -2 ) 

Soluble solids content 

Tirpios sausosios medžiagos (%) 

Skynimai 

average 

average 

2004 2005 

2004 2005 

vidutiniškai 

vidutiniškai 

1 4.3 3.5 3.9 12.2 12.3 12.3 

2 3.9 3.2 3.6 12.1 12.4 12.3 

3 3.7 3.3 3.5 12.0 12.4 12.2 

4 4.2 3.0 3.6 11.9 12.5 12.2 

5 4.0 3.2 3.6 12.0 12.5 12.3 

LSD 05 

/ R 05 

0.24 0.17 0.21 0.20 0.25 0.24 

Weight losses during storage depended on harvest time and were dissimilar every 

year (Fig. 1). In 2004 weight loss was lowest in early picked apples. From 3rd harvest 

its value significantly increased. In 2005 the obviously lowest weight loss was established 

at 3rd harvest. Apples picked at this stage lost by 16–20 % less their mass in 

comparison with earlier or later picked fruits. 

Fig. 1. Effect of harvest time on fruit weight losses during storage 

1 pav. Skynimo laiko įtaka natūraliems masės nuostoliams laikymo metu 

119

On the average up till 19.8 % of apple rotted during 180 days of storage period 

(Fig. 2). The extent of losses linearly depended on harvest time. At each picking the 

significant increase of rotten apples was recorded and the maximum of damaged fruits 

was estimated of apples picked at the latest. At this stage picked apples rotted by 5 times 

more in comparison with apples picked at the earliest harvest. 

Fig. 2. Effect of harvest time on fruit rots incidence during storage 

2 pav. Skynimo laiko įtaka obuolių puvimui laikymo metu 

During the storage ‘Shampion’ apple rot was caused by Monilinia sp., 

Gloeosporium spp. and Penicillium spp. On the average 58 % of apple rots were 

caused by Gloeosporium spp., 21 % by Penicillium spp. and 20 % by Monilinia sp. 

More rot injuries of Gloeosporium rot as dominating rot was detected on latterly 

picked apples. Significantly smaller amount of rotten apples was recorded in apples 

picked at optimum maturity. 

Discussion. During investigation period fruit quality parameters changed according 

to harvest date and dynamic of their changes were similar for each trial year. 

Later harvested fruits were softer and more mature. Fruit storage ability was closely 

connected to fruit maturity too. 

The softening rate of apple fruit vary from cultivar to cultivar, depending on 

the presence and expression of genes, which regulate the activity of hydrolytic enzymes 

(Ingle et al., 2000; Konopacka, Plocharski, 2002; Johnston et al., 2002). In our 

trials, measurements of firmness showed that cv. ‘Shampion’ belong to the group of 

medium firm fruits. Softening rate during ripening and storage was low. Fruits of cv. 

‘Shampion’ tended to lose their firmness slower than ‘Auksis’, ‘Lobo’ and ‘Lodel’ 

(Kviklienė, 2001; Kviklienė et al., 2006), however, faster than fruits of cv. ‘Ligol’ 

(Kviklienė et al., 2008). 

120

The concentration of soluble solids is a good indicator of sugar content and presumably 

of sweetness. Usually later picked apples show higher SSC value not only at 

harvest time, but at the end of storage too (Yong Soo et al., 1998). In our study SSC 

reached maximum at third harvest after which it levelled off. However, post-storage 

SSC was not significantly affected by harvest time. Similar tendencies were obtained 

in different trials (Kviklienė et al., 2006; Wargo, Watkins, 2004; Echeverria et al., 

2002; Braun et al., 1995). 

Loss of mass and decay during storage can greatly affect marketability. Mass loss 

during storage depends on fruit maturity at harvest time (Ferguson et al., 1999). Fruit 

picked at the optimal harvest time lose less mass during storage than fruits picked 

too early or too late (Elgar et al., 1999; Dris and Niskanen, 1999). In our trials lower 

general fruit losses were obtained at 2 nd and 3 rd harvest time. Too early and too late 

picked fruits had bigger losses. 

Incidence of rots and decay in our trial directly depended on harvest time. The 

higher incidence of rots in later picked apples can be explained by more intensive all 

the physiological processes in overmature fruits. Similar results were recorded with 

other apple cultivars (Dris & Niskanen, 1999; Elgar et al., 1999; Ingle et al., 2000; 

Kviklienė, 2004). In both trial years apple fruits were mostly infected by fungus of 

Gloeosporium genus. 

Conclusions. 1. Harvest time has a significant effect on fruit internal quality at 

harvest time and during storage. Apples were of the best quality at the end of storage, 

when maturity index at picking date was 0.22–0.17. 

2. Incidence of decay and rots depended linearly on fruit maturity: more late 

harvest – more fruit loss. 

4. During storage ‘Shampion’ apple rot was caused by Monilinia sp., 

Gloeosporium spp. and Penicillium spp. On the average apple fruits were mostly 

infected by fungus of Gloeosporium genus. 

Gauta 2009 06 30 


References 

1. Braun H., Brosh B., Ecker P., Krumbock K. 1995. Changes in quality off apples 

before, during and after CA-cold storage. Obstau Und Fruchteverwertung, 

45(5–6): 143–206. 

2. Casals M., Bonany J., Carbo J., Alegre S., Iglesias I., Molina D., Casero T., 

Recasens I. 2006. Establishment of a criterion to determine the optimal harvest 

date of ‘Gala’ apples based on consumer preferences. Journal of Fruit and 

Ornamental Plant Research, 14: 53–63. 

3. Dris R., Niskanen R. 1999. Quality changes of ‘Lobo’ apples during cold storage. 

Acta Hort., 485: 125–133. 

121

4. Echeverria G., Graell J., Lopez M. L. 2002. Effect of harvest date and storage 

conditions on quality and aroma production of ‘Fuji’ apples. Food Science and 

Technology International, 8(6): 351–360. 

5. Elgar H. J., Watkins C. B., Lalu N. 1999. Harvest date and crop load effects on 

a carbon dioxide-related storage injury of ‘Braeburn’ apple. HortScience, 34(2): 

305–309. 

6. Ferguson I., Volz R. & Woolf A. 1999. Preharvest factors affecting physiological 

disorders of fruit. Postharvest Biology and Technology, 15: 255–262. 

7. Franelli K., Casera C. 1996. Influence of harvest date on fruit quality and storability 

in the varieties Braeburn and Gala. Cost 94. The postharvest treatment of 

fruit and vegetables. East Malling, 105–115. 

8. Hribar J. Plestenjak A., Simcic M., Vidrih R., Patako. 1996. D. Influence of 

ecological conditions on optimum harvest date in Slovenia. In: A. de Jager, D. 

Johanson, E. Hohn (eds.), The Postharvest Treatment of Fruit and Vegetables. 

Determination and Prediction of Optimum Harvest Date of Apples and Pears. 

COST 94. European Commission. Luxembourg, 49–51. 

9. Ihabi M., Rafin C., Veighie E., Sancholle M. 1998. Storage diseases of apples: 

orchard or in storage. In: First transnational workshop on biological, integrated 

and rational control. Lille, France 21–23 January 1998. Service Regional de la 

Protection des Vegetaux, Nord Pas-de Calais, 91–92. 

10. Ingle M., D’Souza M.C., Townsend E. C. 2000. Fruit characteristics of York 

apples during development and after storage. HortScience, 35(1): 95–98. 

11. Johnston J. W., Hewett E. W., Hertog M. L., Harker F. R. 2002. Harvest date 

and fruit size affect postharvest softening of apple fruit. Journal of Horticultural 

Science & Biotechnology, 77(3): 355–360. 

12. Juan J. L., Frances J., Montesinos E., Camps F., Bonany J. 1999. Effect of 

harvest date on quality and decay losses after cold storage of Golden Delicious 

apples in Girona. Acta Horticulturae, 485: 195–201. 

13. Konopacka D., Plocharski W. J. 2002. Effect of picking maturity, storage technology 

and shelf-life on changes of apple firmness of ‘Elstar’, ‘Jonagold’ and 

‘Gloster’ cultivars. J. Fruit Ornam. Plant Res., 10: 11–22. 

14. Kviklienė N. 2001. Effect of harvest date on apple fruit quality and storage ability. 

Folia Horticulturae, 13(2): 97–102. 

15. Kviklienė N. 2004. Influence of harvest date on physiological and biochemical 

processes in apple fruit. Sodininkystė ir daržininkystė, 23(2): 412–420. 

16. Kviklienė N., Kviklys D., Viškelis P. 2006. Changes in fruit quality during ripening 

and storage in the apple cultivar ‘Auksis’. J. Fruit Ornam. Plant Res., 14(2): 

195–206. 

17. Kviklienė N., Valiuškaitė A., Viškelis P. Effect of harvest maturity on quality 

and storage ability of apples cv. ‘Ligol’. Sodininkystė ir daržininkystė, 27(2): 

339–346. 

18. Rizzolo A., Grassi M., Zerbini P. E. 2006. Influence of harvest date on ripening 

and volatile compounds in the scab-resistant apple cultivar ‘Golden Orange’. 

Journal of Horticultural Science & Biotechnology, 81(4): 681–690. 

122

19. Streif J. 1996. Optimum harvest date for different apples cultivars in the ‘Bodensee’ 

area. In: A. de Jager, D. Johanson, E. Hohn (eds.), The Postharvest Treatment of 

Fruit and Vegetables. Determination and Prediction of Optimum Harvest Date of 

Apples and Pears. COST 94. European Commission. Luxembourg, 15–20. 

20. Vielma M. S., Matta F. B., Silval J. L. 2008. Optimal harvest time of various apple 

cultivars grown in Northern Mississippi. Journal of the American Pomological 

Society, 62(1): 13–20. 

21. Wargo J. M., Watkins C. B. 2004. Maturity and storage quality of ‘Honeycrisp’ 

apples. Horttechnology, 14(4): 496–499. 

22. Yong Soo H., Yong-Pil Ch., Yac Chang L. 1998. Influence of harvest date and 

postharvest treatments on fruit quality during storage and simulated marketing 

in ‘Fuji’ apples. J. of Korean soc. For Hor. Sc., 39(5): 574–578. 

23. Zerbini P. E., Pianezzola A., Grassi M. 1999. Poststorage sensory profiles of fruit 

of apple cultivars harvested at different maturity stages. Journal of Food Quality, 

22(1). 1–17. 


Skynimo laiko įtaka ‘Šampion’ obuolių kokybei vaisiams nokstant ir 

juos laikant 

N. Kviklienė, A. Valiuškaitė 

Santrauka 

2004 ir 2005 m. Lietuvos sodininkystės ir daržininkystės institute tirta veislės ‘Šampion’ 

obuolių, nuskintų skirtingo sunokimo, įtaka jų išsilaikymui. Vaisiai skinti penkis kartus per 

savaitę. Kiekvieno skynimo metu apskaičiuotas sunokimo indeksas. Obuolius laikant matuotas jų 

minkštimo kietumas, tirpių sausųjų medžiagų kiekis ir apskaičiuoti masės nuostoliai. Nustatyta, 

kad vaisių kokybė skynimo ir laikymo metu priklausė nuo sunokimo laipsnio. Vėliau nuskinti 

vaisiai buvo minkštesni ir skynimo metu, ir laikymo pabaigoje. Metų įtaka vaisių kokybės 

rodikliams buvo nežymi. Laikymo pabaigoje geriausia kokybe pasižymėjo vaisiai nuskinti, 

kai sunokimo indeksas skynimo metu buvo 0,22–0,17. Sandėliavimo metu ‘Šampion’ veislės 

obuoliai buvo pažeisti Monilinia sp., Gloeosporium spp. ir Penicillium spp. genčių grybų. 

Abejais tyrimų metais dominavo Gloeosporium spp. genties grybų sukeliami obuolių puviniai. 

Reikšminiai žodžiai: laikymas, Malus × domestica, masės nuostoliai, puviniai, sunokimo 

indeksas. 

123




Incidence of fruit rot on strawberries in Latvia, 

resistance of cultivars and impact of cultural systems 

Valda Laugale 1 , Līga Lepse 1 , Līga Vilka 2 , Regīna Rancāne 2 

1 

Pure horticultural Research Centre, Abavas 2, Pure, Tukuma r., LV-3124 

Latvia, e-mail pures_dpc@tukums.parks.lv 

2 

Latvian Plant Protection Research Center, Lielvardes 36/38, Riga, LV-1006 

Latvia, e-mail regina.rancane@laapc.lv 

In 2007 and 2008 strawberry plantations in different regions of Latvia were inspected 

looking for fruit rots. Causal agents of fruit rot were detected at the laboratory of Latvian 

Plant Protection Research Center. 28 strawberry plantations were inspected in 2007. In this 

year weather conditions during strawberry flowering and harvest time were not favorable for 

the development of the diseases. On the damaged fruits and flowers mostly fungus Botrytis 

cinerea was detected at laboratory. On some fruits rots caused by Hainesia lynthri, Muccor spp. 

and Penicillium spp. were found. Next year 26 strawberry plantations were inspected. Botrytis 

cinerea was detected on samples with damages like pale brown fruit rot, dead flowers and 

ovaries and dark brown spots on pedicles. Causal agents Hainesia lynthri, Phomopsis obscurans, 

Coniella castaneicola, Fusarium spp., Muccor spp., Rhizopus spp., Penicillium spp. on 

rotted fruits also were found at laboratory. 

Susceptibility to Botrytis rot of 16 strawberry cultivars was evaluated at the Pure HRC 

in 2006–2008. On the average during three production years, cultivars ‘Honeoye’ and ‘Tenira’ 

had the lowest Botrytis incidence, but ‘Venta’ and ‘Bounty’ were the most susceptible among 

the tested cultivars. In 2008, experiments on extending of strawberry production season 

using different plant covers, plastic soil mulches, cultivars, and “frigo” plants were started at 

the Pure HRC. The first results showed the significant effect of plastic soil mulches and plant 

covers on reduction of Botrytis incidence. 

Key words: cultivar susceptibility, cultural practices, Fragaria × ananassa, grey mold, 

incidence. 

Introduction. Strawberry is one of the most popular commercial berry crops in 

Latvia. In 2007 the area of strawberries was about 500 ha. Most of production is used 

for fresh local markets. The yield is quite low in comparison with other European 

countries, on the average 3–5 t ha -1 . There are several reasons for it. Mostly yield 

loses can be caused both by unfavourable environment conditions and due to damage 

by different pathogenic organisms. The efficiency of plant protection is low in many 

farms. Grey mould, leaf spots, mildew, root diseases and nemathods are mentioned 

as the most widespread on strawberries under Latvia conditions (Moročko, 2003; 

125

Laugale et al., 2004). Especially high yield loses can be caused by fruit rots. The 

implementation of Integrated Plant Protection system demands to reduce the using of 

chemical plant protection products and to look for alternative methods like more resistant 

cultivars and proper agriculture (Meszka and Bielenin, 2004). Studies described 

here were started with aim to determine the main causal agents for strawberry fruit 

rots in Latvia and to evaluate the susceptibility of widely grown cultivars. The first 

investigation results revealing the influence of cultural practice on fruit rot reduction 

in strawberry fields also are presented. 

Object, methods and conditions. 28 strawberry plantations in 2007 and 26 

plantations in 2008 were inspected for fruit rot detection in different Latvia regions 

(Fig. 1). In 2007 the amount of fruit rot samples was lower than in 2008 because weather 

conditions during strawberry flowering and harvest time were optimal for fruit 

formation, but not favourable for rot development. During the harvest time samples 

of rotted fruits, died off flowers, ovaries and dark brown pedicles were collected for 

causal agent detection. Causal agents of strawberry rot were isolated in pure culture, 

used PDA (potato-dextrose agar). Discovered fungi were identified at the laboratory 

of Latvian Plant Protection Research Center. 

Fig. 1. Inspected strawberry plantations in Latvia, 2008 

1 pav. Tikrintos braškių plantacijos Latvijoje, 2008 m. 

16 widely grown in Latvia strawberry cultivars were evaluated on susceptibility 

to Botrytis rot at the Pure Horticultural Research Station in 2006–2008. Plants were 

planted in biologically certified field in single rows at spacing of 30 × 100 cm in the 

autumn of 2005. A completely randomized block design with four replicates and thirty 

plants per plot was used. The following cultivars were tested: ‘Bogota’, ‘Bounty’, 

126

‘Dukat’, ‘Feierverk’, ‘Honeoye’, ‘Induka’, ‘Jonsok’, ‘Rubinovij Kulon’, ‘Pandora’, 

‘Polka’, ‘Senga Sengana’, ‘Symphony’, ‘Siurprise Olimpiadi’, ‘Tenira’, ‘Venta’ and 

‘Zefyr’. Only in organic farming permitted plant protection and nutrition products 

were used. 

In 2008 at the Pure Horticultural Research Center the damage by fruit rots were 

evaluated in following trials: 1) 3 cultivars grown on soil mulched with black plastic 

and no soil mulch; in spring till the beginning of production plants were covered with 

transparent film (polyethylene) or agronet (Agryl 17) and without cover as control; 

2) “frigo” plants (3 cultivars) in two planting densities (3.3 and 6.6 plants m -2 ) grown 

on soil mulched with white plastic (black lower side) and no mulch; 3) two plant 

types M 0 

and M 1 

generation from micropropagation of everbearer cultivar ‘Brighton’ 

grown on soil mulched with white plastic (black lower side) and no mulch. In all trials 

plants were spaced 40 cm apart within rows (except one variant in trial with “frigo” 

plants where 20 cm distance between plants also was used), and 30 cm apart between 

rows. The beds were 150 cm apart, center-to-center. Split-split block design with four 

replicates was used. No fungicides were applied. 

In all trials rotted berries were harvested and weighted at each picking time separately, 

and the percentage of damaged berries from total yield was calculated. The data 

were analyzed using descriptive statistics and ANOVA (probability 95 %). Duncan’s 

multiple range test was applied to compare the means. 

Results. I n c i d e n c e o f f r u i t r o t i n s t r a w b e r r y p l a n t a t i o n s. 

In 2007 from 28 inspected strawberry plantations 51 samples of damaged fruits were 

collected. Pale brown rot spots were the most widespread damages on unripe and ripe 

fruits. Some flowers were died off as well. Botrytis cinerea was detected on damaged 

fruits and flowers in 57 % of inspected strawberry farms. Only in four plantations 

in West and one in South region of Latvia ripe fruit rot caused by Muccor spp. was 

detected. In few plantations rot caused by Hainesia lynthri, Fusarium spp., Rhizopus 

spp. and Penicillium spp. (Maas, 1998) also were found. 

In 2008 from 26 strawberry plantations 127 samples of rotted strawberry were 

collected. The pale brown fruit rot, died off flowers, ovaries and dark brown spots 

on pedicles were the main damages in strawberry plantations. Botrytis cinerea was 

detected in all inspected plantations (Fig. 2). Infection level of Botrytis cinerea was 

very high, because weather conditions were optimal for fungus progression. 

Powdery mildew (Sphaerotheca macularis) on fruits was observed only in 

Kurzeme region in 44 % of the inspected fields. Fusarium spp. developed at laboratory 

from samples with fruits, dead flowers, ovaries and dark brown pedicles. The fungus 

usually causes the rot of root; in this case probably infection level was very high and 

conidia by splash got from soil to flowers, and fruits. Fusarium spp. was detected in 

44 % of the plantations in Kurzeme and in 25 % in Vidzeme (Fig. 3). In samples with 

ripe fruit rot Mucor spp. and Rhizopus spp. were found (Smith et al., 1979). 

127

Fig. 2. Spread of Botrytis cinerea on strawberry samples taken from 

different places in Latvia, in 2007 and 2008 (%) 

2 pav. Botrytis cinerea paplitimas braškių pavyzdžiuose, 

paimtuose skirtingose Latvijos vietovėse 2007 ir 2008 metais, % 

Fig. 3. Spread of causal agents on strawberry samples taken from 

different places in Latvia, in 2008 (%) 

3 pav. Ligos sukėlėjų paplitimas braškių pavyzdžiuose, 

paimtuose skirtingose Latvijos vietovėse 2008 metais, % 

Causal agents Hainesia lynthri, Phomopsis obscurans, Coniella castaneicola and 

Penicillium spp. were detected only in few plantations in Kurzeme. The detection of 

the fruit rot causal agents will be continued. 

S u s c e p t i b i l i t y o f c u l t i v a r s. In strawberry cultivar trial the spreading 

of fruit rots was very low in 2006. In 2007, the percentage of damaged fruits increased, 

but did not exceed 7 % from total yield. Late ripening cultivars were more damaged 

than early ripening, because there was increased amount of precipitation at the end of 

128

season. The lowest percentage of damaged berries had cultivars ‘Honeoye’, ‘Zefyr’ 

and ‘Rubinovii Kulon’ (data not shown). Cultivar ‘Bounty’ had the highest percentage 

of damaged fruits. In 2008, the amount of rotted fruits on the average was significantly 

higher than in 2006 and 2007. It varied from 1.1 to 7.2 % from total yield depending on 

cultivar (data not shown). Cultivars ‘Honeoye’ and ‘Tenira’ had the lowest percentage 

of damaged berries, but ‘Venta’ was the most susceptible between tested cultivars. 

On the average of three production years, cultivar ‘Honeoye’ had the lowest fruit 

rot incidence (Fig. 4). Cultivars ‘Tenira’, ‘Jonsok’, ‘Rubinovij Kulon’ and ‘Zefyr’ also 

showed low susceptibility. 

Fig. 4. The amount of rotted fruits (%) from total yield on the average of 

three production seasons in strawberry cultivar trial. 

Values in columns followed by the same letter do not differ significantly 

according to Duncan’s multiple range test (p = 0.05) 

4 pav. Vidutinis supuvusių vaisių kiekis (%) nuo bendro derliaus 

braškių veislių bandyme per trejus derėjimo metus. 

Stulpeliai, pažymėti vienodomis raidėmis, 

patikimai nesiskiria pagal Dunkano kriterijų (p = 0,05) 

129

‘Venta’ and ‘Bounty’ had the highest percentage of rotted fruits on the average 

of three investigation years between tested cultivars. 

Impact of cultural practices. In the trial for early production using black plastic 

soil mulch and different plant covers the significant differences in the percentage of 

rotted fruits was stated, while the amount of damaged berries was low in 2008 (0–2.4 % 

from total yield depending on treatment). The percentage of rotted fruits was significantly 

lower (p = 0.007) than this of plants grown on black plastic mulch comparing 

to non-mulched soil (Fig. 5). 

Fig. 5. The amount of rotted fruits (%) from total yield in 2008 trial using 

black plastic soil mulch and different plant covers for early production. 

Columns assigned by different letters are significantly different, when p ≤ 0.05 

5 pav. Supuvusių vaisių kiekis (%) nuo bendro derliaus 2008 metų bandyme, 

panaudojant juodą plastikinį dirvos mulčią ir skirtingas augalų priedangas ankstyvai produkcijai. 

Stulpeliai, pažymėti skirtingomis raidėmis, patikimai skiriasi, kai p ≤ 0,05 

Notably low amount of rotted fruits was obtained from plants grown with polyethylene 

cover. Agronet cover also significantly reduced the percentage of rotted 

fruits comparing to control (without any plant cover) (Fig. 5). The highest percentage 

of rotted fruits was observed on plants grown without any soil mulch and plant covering, 

but the lowest one – on plants grown on beds with black plastic soil mulch and 

polyethylene cover. 

In the trial where “frigo” plants in two planting densities and white with black 

lower side plastic soil mulch were used, significantly lower percentage of rotted fruits 

was obtained from plants grown on plastic mulch (p = 0.03) comparing to non-mulched 

treatment (Fig. 6 ). 

Significant difference in the percentage of rotted fruits between two planting 

densities was not stated in this year (Fig. 6). There was observed tendency for increasing 

of Botrytis damage in higher plant density for plants grown on beds with plastic 

mulch. 

130

Fig. 6. The amount of rotted fruits (%) from total yield in trial 2008 with “frigo“ plants in 

two planting densities and white with black lower side plastic soil mulch for late production. 

Columns assigned by different letters are significantly different, when p ≤ 0.05 

6 pav. Supuvusių vaisių kiekis (%) nuo bendro derliaus 2008 metų bandyme su “frigo” augalais, pasodintais 

dvejopu tankumu, ir panaudojant baltą su juoda apatine puse plastikinį dirvos mulčią vėlyvai 

produkcijai. Stulpeliai, pažymėti skirtingomis raidėmis, patikimai skiriasi, kai p ≤ 0,05 

Fig. 7. The amount of rotted fruits (%) from total yield in 2008 trial with everbearer strawberry 

cultivar ‘Brighton’ using two plant types and white with black lower side plastic soil 

mulch. Columns assigned by different letters are significantly different, when p ≤ 0.05 

7 pav. Supuvusių vaisių kiekis (%) nuo bendro derliaus 2008 metų bandyme su nuolat derančia braškių 

veisle ‘Brighton’ panaudojant du augalų tipus ir baltą su juoda apatine puse plastikinį dirvos mulčią. 

Stulpeliai, pažymėti skirtingomis raidėmis, patikimai skiriasi, kai p ≤ 0.05 

131

In the trial with everbearer cultivar ‘Brighton’ using two plant types and white 

with black lower side plastic soil mulch, significantly lower percentage of rotted fruits 

was obtained from plants grown on plastic mulch (p = 0.03) comparing to non-mulched 

treatment (Fig. 7). 

Significant difference in the percentage of rotted fruits between two plant types 

M 0 

and M 1 

was not stated (Fig. 7). 

Discussion. In our investigations Botrytis cinerea was determined as the main 

casual agent of strawberry fruit rotting. It conform to the previous investigations 

carried out in Latvia (Dūks, 1976; Moročko, 2003; Laugale et al., 2004). Botrytis rot 

is also one of the most important diseases of strawberry, reducing yield and quality 

pre- and post-harvest in all strawberry production areas (Berrie et al., 2000; Legard 

et al., 2002; Rigotti, Viret, 2004). It is important to remove rotted fruits from the field 

during the harvest time to reduce infection level in strawberry plantations. 

In the evaluation of strawberry cultivars significant difference in susceptibility 

to grey mould between cultivars was stated. Only cultivar ‘Venta’ had significantly 

higher percentage of damaged fruits than ‘Senga Sengana’, which is characterized as 

susceptible to grey mold (Zurawicz, Daubeny, 1995). Several cultivars like ‘Bounty’, 

‘Siurprise Olimpiadi’, ‘Feierverk’, ‘Pandora’, ‘Dukat’ and ‘Bogota’ showed susceptibility 

similar to this of ‘Senga Sengana’. In previous investigations in Pure cultivars 

‘Dukat’ and ‘Bogota’ showed good resistance to Botrytis rot (Laugale et al., 2004; 

Laugale, Lepse, 2007). Also in this trial they had lower percentage of rotted fruits 

than ‘Senga Sengana’, but the difference was not significant. It can be explained by 

overall low spreading of the rots during test years 2006–2008. ‘Honeoye’ was the 

most resistant to grey mould between tested cultivars. It showed also good resistance 

to Botrytis rot in Finland (Matala, 2002), Poland (Cieśliński et al., 1993) and Estonia 

(Libek, 1997). 

In all trials where different cultural practices were applied the significant impact 

of plastic soil mulches both black, and white with black lower side on the reduction 

of fruit rot incidence was stated. The positive effect of black plastic soil much on 

reducing of grey mold damage on strawberries is reported also by Lille et al. (2003), 

Plekhanova and Petrova (2002). Using plant covers, especially polyethylene film, till 

the beginning of harvesting also reduced the percentage of rotted fruits. According to 

Xiao et al. (2001), shorter periods of leaf wetness and higher temperatures in plastic 

tunnels may have contribute to a lower incidence of Botrytis rot on fruit in comparison 

to the open field. According to the investigations in Finland, Agronet plant cover 

reduced different disease spreading, and succeeded to stop the spreading of grey mold 

on late cultivar ‘Hiku’ (Dalman, 1993). The increase of planting density did not cause 

significant increase of Botrytis rot damage in our trial. Probably because the fruit rot 

incidence was low in this year and in the first cropping year plants were not fully-grown. 

According to the investigations of Daugaard (2003), climatic factors may play more 

important role in the control of Botrytis than differences in plant density. 

Conclusions. The main causal agent of strawberry fruit rotting in Latvia strawberry 

plantations is Botrytis cinerea Pers. It causes pale brown fruit rot, death of 

flowers and ovaries and dark brown spots on pedicles. Mucor spp. and Rhizopus spp. 

were the main casual agents for ripe (post-harvest) rot. 

132

Strawberry cultivars grown in Latvia have significantly different resistance level 

to Botrytis rot. Cultivars ‘Honeoye’ and ‘Tenira’ showed the highest resistance, but 

‘Venta’ and ‘Bounty’ were the most susceptible among the tested cultivars. 

According to the first obtained results, plastic soil mulches and plant covers can 

significantly reduce fruit rot incidence. 

Acknowledgements. Theses studies were funded by the Latvian Ministry of 

Agriculture. 

Gauta 2009 06 30 


References 

1. Berrie A. M., Harris D. C., Xiangming Xu., Burgess C. M. 2000. A system for 

managing Botrytis and powdery mildew of strawberry: first results. IOBC WPRS 

Bulletin, 23(11): 35–40. 

2. Cieśliński G., Klimczan A., Smolarz K. 1993. Field performance of some new 

American and polish strawberry cultivars grown in Poland. Acta Horticulturae, 

348: 171–176. 

3. Dalman P. 1993. Polypropylene row cover in pesticide free production of strawberry 

in Finland. Acta Horticulturae, 348: 489–492. 

4. Daugaard H. 2003. Effect of plant spacing on yield and incidence of Botrytis 

cinerea Pers. in strawberry. IOBC WPRS Bulletin, 26(2): 147–151. 

5. Dūks V. 1976. Zemenes. Liesma. Rīga, 170. 

6. Laugale V., Lepse L. 2007. Research trials on strawberry cultivars in Pūre 

Horticultural Research Station (Latvia) during the last 10 years. Sodininkystė ir 

daržininkystė, 26(3): 81–92. 

7. Laugale V., Morocko I., Petrevica L. 2004. Problems for strawberry culture in 

Latvia. IOBC/WPRS Bulletin, 27(4): 37–40. 

8. Legard D. E., Mertely J. C., Xiao C. L., Chandler C. K., Duval J. R., Price J. P. 

2002. Cultural and chemical control of Botrytis fruit rot of strawberry in annual 

winter production systems. Acta Horticulturae, 567: 651–654. 

9. Libek A. 1997. Strawberry cultivar trials at the Polli Horticultural Institute. 

Sodininkystė daržininkystė, 26(3): 160–163. 

10. Lille T., Karp K., Värnik R. 2003. Profitability of different Technologies of 

strawberry cultivation. Agronomy Research, 1: 75–83. 

11. Maas J. L. 1998. Compendium of Strawberry Diseases. Second edition published. 

The American Phytopathological Society, 17–62. 

12. Matala V. 2002. Strawberry variety trials on berry farms. Acta Horticulturae, 

567(1): 215–217. 

13. Meszka B., Bielenin A. 2004. Possibilities of integrated grey mold control on 

strawberry plantations in Poland. IOBC/WPRS Bulletin, 27(4): 41–45. 

133

14. Moročko I. 2003. Ogulāju slimības. In: Bankina B. (ed.) Augu slimības. Latvijas 

Lauksaimniecības Universitāte. Jelgava, 206–227. 

15. Plekhanova M. N., Petrova M. N. 2002. Influence of black plastic soil mulching 

on productivity of strawberry cultivars in Northwest Russia. Acta Horticulturae, 

567: 491–494. 

16. Rigotti S., Viret O. 2004. Fungal flora in strawberry plants and relative importance 

of Botrytis cinerea. IOBC/WPRS Bulletin, 27(4): 47–53. 

17. Smith W. L., Moline H. E., Johnson K. S. 1979. Studies with Mucor species 

causing postharvest decay of fresh produce. Phytopathology, 69: 865–869. 

18. Xiao C. L., Chandler C. K., Price J. F., Duval J. R., Mertely J. C., Legard D. E. 

2001. Comparison of epidemics of Botrytis fruit rot and powdery mildew of 

strawberry in large plastic tunnel and field production systems. Plant Disease, 

901–919. 

19. Zurawicz E., Daubeny H. 1995. ‘Senga Sengana’ strawberry. Fruit Varieties J., 

49: 130–132. 


Vaisių puvinio paplitimas ant braškių Latvijoje, veislių atsparumas ir 

kultūrinių sistemų įtaka 

V. Laugale, L. Lepse, L. Vilka, R. Rancāne 

Santrauka 

2007 ir 2008 metais įvairiuose Latvijos regionuose patikrintos braškių plantacijos ieškant 

vaisių puvinių. Vaisių puvinio sukėlėjai nustatyti Latvijos augalų apsaugos tyrimų centro laboratorijoje. 

2007 metais patikrintos 28 braškių plantacijos. Tais metais oro sąlygos braškių žydėjimo 

ir derliaus metu nebuvo palankios ligų vystymuisi. Ant pažeistų vaisių ir žiedų laboratorijoje 

daugiausia aptiktas Botrytis cinerea grybas. Ant kai kurių vaisių rasta Hainesia lynthri, Muccor 

spp. ir Penicillium spp. sukeliamų puvinių. Kitais metais patikrintos 26 braškių plantacijos. 

Botrytis cinerea buvo aptiktas ant pavyzdžių su tokiais pažeidimais, kaip šviesiai rudas vaisių 

puvinys, negyvi žiedlapiai bei mezginės ir tamsiai rudos dėmės ant žiedkočių. Be to, ant puvinio 

pažeistų vaisių laboratorijoje rasti šie ligos sukėlėjai – Hainesia lynthri, Phomopsis obscurans, 

Coniella castaneicola, Fusarium spp., Muccor spp., Rhizopus spp., Penicillium spp. 

16 braškių veislių jautrumas Botrytis puviniui buvo įvertintas Pure LTC 2006–2008. Per 

trejus derėjimo metus veislės ‘Honeoye’ ir ‘Tenira’ buvo mažiausiai užsikrėtusios Botrytis, o 

‘Venta’ ir ‘Bounty’ – iš visų tirtų veislių labiausiai. 2008 metais Pure LTC pradėti tyrimai, kaip 

pratęsti braškių derėjimo sezoną, panaudojant įvairias augalų priedangas, plastikinius dirvos 

mulčius, veisles ir “frigo” augalus. Pirmieji rezultatai parodė reikšmingą plastikinių dirvos 

mulčių ir augalų priedangų poveikį Botrytis paplitimo sumažinimui. 

Reikšminiai žodžiai: agrotechninės priemonės, Fragaria × ananassa, kekerinis puvinys, 

paplitimas, veislės jautrumas. 

134




Prevalence peculiarities of airborne Alternaria genus 

spores in different areas of Lithuania 

Rita Mikaliūnaitė, Martynas Kazlauskas, Laura Veriankaitė 

Department of Environmental Research, Nature Science Faculty, Šiauliai University, 

P. Višinskio 19-115, Šiauliai, LT-77156, Lithuania, e-mail oikos@fm.su.lt 

The annual, seasonal and hourly distribution of Alternaria Ness. spores in the air was 

measured in three urban areas in Lithuania – Klaipėda, Šiauliai, and Vilnius in 2005–2006. 

Hirst type 7-day recording spore traps were used for spore fixation. Fungal spores were identified 

and counted according to 12 transverse traverse method. Duration of spore season was 

determined using 90 % method. Duration of season ranged from 62 days in Vilnius in 2005 to 

97 days in Klaipėda in 2006. The biggest amount of Alternaria spores was established in summer 

and the least one – in winter and spring. Only solitary spores were observed in February, 

March and April in all aerobiological stations. The total annual amount of Alternaria reached 

3 090 spores in Vilnius aerobiological station in 2005 and 9 718 spores in Klaipėda in 2006. 

The peak of sporification season was recorded in August in all locations and it was from 121 

to 707 spores m -3 . In this month there were observed 51.4–70.3 % of all annual spores counted. 

The hourly pattern of Alternaria spores concentration in August indicated maximum value 

between 13:00 and 1:00 hours. Minimal amounts were recorded at 5:00–9:00 hours. 

Key words: Alternaria, fungal spores, season duration, spore concentration. 

Introduction. Alternaria is an extremely common and widespread saprophyte 

or plant pathogenic genus, which causes big economic loses throughout the world 

(Corden et al., 2003; Munuera et al., 2001). The analysis of the prevalence of this 

fungus spores is substantial, because Alternaria colonises all the stages of plant 

growth (Mitakakis et al., 2001). It’s necessary to emphasize that some species are 

plant pathogens, which collectively cause a range of diseases with impact on a large 

variety of important agronomic host plants including cereals, ornamentals, oilcrops, 

vegetables and fruits (Thomma, 2003). Common Alternaria micromycetes A. brassicicola, 

A. brassicae, A. raphani, A. alternata cause diseases of oilseed and turnip 

rapes (Petraitienė, 2005), A. radicina, A. dauci – of carrots (Survilienė, Valiuškaitė, 

2006), A. brassicicola and A. brassicae – of cabbages (Survilienė et al., 2004), besides, 

A. alternata, A. radicina and A. tenussisima cause human diseases also (Lugauskas 

et al., 2002). 

Most aeromycology investigations are conducted in towns (Corden et al., 2003; 

Kasprzyk et al., 2004; Stępalska, Wołek, 2005; Kasprzyk, Worek, 2006); however, some 

135

esearchers pay attention to data originating from countryside (Mitakakis et al., 2001; 

Brazauskiene, Petraitiene, 2006; Kasprzyk, Worek, 2006). In rural environments the 

spread of airborne fungal spores there was investigated to determine occurrence during 

the crop harvest (Mitakakis et al., 2001) or to estimate the quantity of plant pathogenic 

fungal spores in the air (Brazauskiene, Petraitiene, 2006). During these studies fungal 

spores were sampled with Burkard volumetric spore traps, which were located in the 

fields of crops (Brazauskiene, Petraitiene, 2006). Passive sedimentation plates are the 

alternative method, which was used to evaluate the concentration of fungal spores in 

the air. This method is more practical since allows to estimate viable micromycetes 

spores and to identify species. Passive sedimentation plate method is being used for 

the short time (Lugauskas et al., 2003), while the volumetric method allows evaluating 

the season of fungal spores (Stępalska et al., 1999; Corden, Millington, 2001; 

Corden et al., 2003; Konopińska, 2004; Oliveira et al., 2005; Kasprzyk, Worek, 2006). 

In Lithuania passive method is the most common. The most common micromycetes 

species in Vilnius during various seasons of 1996–1999 was Alternaria alternata 

(Lugauskas et al., 2003). 

The aim of this work is to study the annual, seasonal and hourly distribution of 

Alternaria airborne spores in the air of three cities in Lithuania. This comparative study 

of coastal (Klaipėda) and landlocked (Vilnius and Šiauliai) areas of Lithuania is especially 

interesting, since it allows evaluating the peculiarities of spore dispersion. 

Objects, methods and conditions. Spores were sampled at three sites, in different 

places, which partly represented Lithuania – in Klaipėda (Western Lithuania, 

coastal place), Šiauliai (Northern Lithuania, landlocked place), and Vilnius (Southern 

Lithuania, landlocked place). Monitoring was carried out using three volumetric 

Hirst-type 7-day recording spore traps with flow rate of 10 L min -1 . Spore traps were 

located in Klaipėda – 55°45′20 N and 21°07′32 E, in Šiauliai – 55°55′36 N and 

23°18′ E, in Vilnius – 54°40′40 N and 25°16′05 E. 

The measurements of airborne fungal spore concentration in the air were carried 

out by volumetric method in 2005–2006 (Lacey, West, 2006). Spore monitoring took 

place in Šiauliai from 4 of February to 31 of December in 2005 and from 22 of March 

to 11 of October in 2006. In second site, Klaipėda it took place from 28 of January to 

26 of September in 2005 and from 8 of March to 12 of October in 2006. In Vilnius, 

it took place from 3 of February to 22 of September in 2005 and from 9 of March to 

30 of September in 2006. Melinex tape, greased with a thin layer of silicone solution 

was changed once in a week at the same time (Mandrioli et al., 1998). The exposed 

tape was cut in 48 mm segments, thus each represented 24 hours period. Average daily 

concentrations (spores m -3 ) were obtained by counting spores for every two hours by 

twelve transverse traverse method (Sterling et al., 1999). Sample representing each 

day were analysed for spores of Alternaria by light microscope Nikon Eclipse 50i at 

a magnification of 400x (0.50 mm microscopic field). For microscopic fungal spore 

identification Lacey, West (2006) and Käärik et al. (1983) publications were used as 

reference books. 

Estimated spore values were transformed into the number of spores in each cubic 

metre of air sampled per day. The duration of spore season was calculated by 90 % 

136

method – the start of Alternaria spore season was defined as the date when 5 % of 

the seasonal cumulative spore amount was trapped, and the end of the season – as the 

date when 95 % of the seasonal cumulative spore amount was reached (Stępalska, 

Wołek, 2005; Kasprzyk, Worek, 2006). The days when Alternaria spore amount 

reached 100 spores in cubic meter of air were recorded as the peak days. The hourly 

distribution of Alternaria spores concentration was determined in August, the peak 

of spore season. Statistical correlation and regression analysis was carried out using 

Stat-Eng programme. Significant difference was assessed at a critical value of p < 

0.05 and p < 0.01. 

Results. Seasonal data of Alternaria s p o r e s. The duration of 

spore season, calculated by 90 % method is shown in Table 1. 

Table 1. Seasonal data of Alternaria spores in Lithuania in 2005–2006 

1 lentelė. Alternaria sporų sezono trukmė Lietuvoje 2005–2006 m. 

Year 

Metai 

Start date 

Pradinė data (5 %) 

2005 12 July 

Liepos 12 

2006 19 June 

Birželio 19 

2005 20 July 

Liepos 20 

2006 11 July 

Liepos 11 

2005 12 July 

Liepos 12 

2006 21 June 

Birželio 21 

End date 

Galinė data (5 %) 

Klaipėda 

14 September 

Rugsėjo 14 

23 September 

Rrugsėjo 23 

Šiauliai 

6 October 

Spalio 6 

18 September 

Rugsėjo 18 

Vilnius 

11 September 

Rugsėjo 11 

18 September 

Rugsėjo 18 

Duration (days) 

Trukmė (dienomis) 

65 

97 

78 

68 

62 

90 

The Alternaria spore season duration range was from 62 days in Vilnius in 2005 

to 97 days in Klaipėda in 2006. In both years of research the most stable duration of 

spore season was in Šiauliai, it ranged to 68–78 days. In both years of observation spore 

season appeared to begin at the earliest in Klaipėda and Vilnius, whereas in Šiauliai 

it was late for 8 days in 2005 and 20–22 in 2006 (depending on stations). The end of 

Alternaria spore season during monitoring period in all aerobiological stations occurred 

in September, except Šiauliai in 2005, where spore season occurred in October. 

Annual and monthly totals of Alternaria s p o r e s. The cumulative 

annual and monthly totals of daily average concentrations of Alternaria spores 

are shown in Table 2. The highest Alternaria monthly concentrations were observed 

at the end of summer, in the middle of spore season in all three locations in both years 

of study period. Spore sum in August constituted 51.4–65.6 % of all spores counted in 

137

2005, and 56.4–70.3 % of all spores counted in 2006. In June and May, irrespectively 

the season, Alternaria spores occurred in the air at low concentrations and sporadically 

in March, April and October in all sites. A strong 95 % significant positive correlation 

(r = 0.942) was obtained between the monthly Alternaria spore amounts in different years 

in Klaipėda. Similar findings were in the rest two aerobiological stations: a strong 99 % 

positive correlation (r = 0.992) in Šiauliai and (r = 0.962) in Klaipėda. 

Table 2. Annual and monthly totals and the highest peak day concentrations of 

Alternaria spores in Lithuania in 2005–2006, (peak values in brackets) 

2 lentelė. Alternaria sporų pasiskirstymas pagal metus bei mėnesius ir piko dienos koncentracijos 

Lietuvoje 2005–2006 m. (piko duomenys sliausteliuose) 

Year 

Metai 

Monthly total amount (spores m -3 ) / 

Highest concentration on peak day (spores m -3 ) / 

Bendras kiekis per mėnesį, sporos m- 3 / 

Didžiausia piko dienos koncentracija, sporos m -3 

Annual 

totals (spores 

m -3 ) 

Iš viso per 

metus, 

sporos m -3 

May 

gegužė 

June 

birželis 

July 

liepa 

August 

rugpjūtis 

September 

rugsėjis 

Klaipėda 

2005 29 / 5 / 61 / 13 / 903 / 124 / 2 674 / 306 / 796 / 74 / 4 476 

2006 124 / 11 / 348 / 67 / 1 832 /208 / 5 888 / 707 / 1 253 / 135 / 9 718 

Šiauliai 

2005 39 / 8/ 66 / 10 / 735 / 126 / 4 038 / 375 / 868 / 82 / 6 160 

2006 37 / 5 / 109 / 18 / 1 001 / 139 / 4 205 / 547 / 568 / 61 / 5 984 

Vilnius 

2005 26 / 4 / 77 / 13 / 1 008 / 220 / 1 588 / 121 / 386 / 65 / 3 090 

2006 95 / 16 / 267 / 30 / 1 365 / 249 / 3 511 / 414 / 876 / 122 / 6 223 

The highest daily concentration of Alternaria spores was recorded in the middle 

of the season (August) in Klaipėda and Šiauliai aerobiological stations in both years. 

The highest concentrations occurred at the beginning of spore season only in Vilnius 

in 2005 and in Šiauliai in 2006. 

The number of peak days and maximal concentration of Alternaria spores in 

aerobiological stations are shown in Table 3. Alternaria spores had their monthly 

peaks in July and August in all aerobiological stations. Concentration range in 2005 

was from 121 to 306 spores m -3 air, while in 2006 concentration range was considerably 

higher – from 122 to 707 spores m -3 air. 

No peak days in September of 2005 were recorded, in the same time in 2006 – 

only few. Only 5 days were recorded as peak days of Alternaria spores in Vilnius in 

2005, among those 3 days in July at the start of spore season. Nevertheless, in the 

second monitoring year there were recorded 14 spore day peaks in this aerobiological 

station. The biggest number of spore peak days was determined in Klaipėda in 

2006 – even 28 days. 

138

Table 3. Number of Alternaria spore peak days and concentration of peak day 

in Lithuania in 2005–2006 

3 lentelė. Alternaria sporų piko dienų kiekis bei piko dienų koncentracijos Lietuvoje 

2005–2006 m. 

Aerobiological 

station 

July 

Liepa 

August 

Rugpjūtis 

September 

Rugsėjis 

Aerobiologinė 

stotis 

2005 year 

2005 metai 

2006 year 

2006 metai 

2005 year 

2005 metai 

2006 year 

2006 metai 

2005 year 

2005 metai 

2006 year 

2006 metai 

Number of days with ≥ 100 spores in m -3 

Dienų skaičius, kai ≥ 100 sporų m -3 

Klaipėda 3 6 8 18 - 4 

Šiauliai 1 2 13 17 - - 

Vilnius 3 3 2 10 - 1 

Maximal concentration of spores on peak day in m -3 

Maksimali sporų koncentracija piko dieną, m -3 

Klaipėda 124 208 306 707 - 135 

Šiauliai 126 139 375 547 - - 

Vilnius 220 249 121 414 - 122 

A strong 95 % no significant positive correlation (r = 0.956) was obtained between 

the maximal concentrations of spores on peak day every month in Klaipėda in different 

years. The similarly findings were in other aerobiological station: a strong 99 % positive 

no significant correlation (r = 0.996) in Šiauliai and medium (r = 0.485) in Vilnius. 

Hourly pattern of Alternaria s p o r e s. Maximum value of spore concentration 

in August was found in midnight, between 23:00–1:00 in Klaipėda in 2005, 

but next year of monitoring the maximal values were established between 17:00–19:00. 

Minimal spore counts were noted in 5:00–7:00 in both years of investigations. 

The hourly analysis of samples corresponding air of Šiauliai in August in 2005 

demonstrated the highest concentrations in the afternoon, 13:00–15:00. In the second 

year of monitoring maximal values were determined between 17:00–19:00. The lowest 

Alternaria spore amounts here reached at night between 3:00–7:00 in 2005 and 

2006. 

In Vilnius aerobiological station clear daily peak of spore concentration in August 

wasn’t recognized, concentration of spores was equally dispersed and only obscure 

increase was noticed in 19:00–3:00 time interval. The least amount of spores was 

counted in the morning at 9:00. In the second year there were differences: maximal 

values appeared to be between 17:00 and 19:00, while minimal at 5:00–7:00. 

Roughly, the highest concentrations of Alternaria spores were between 15:00 and 

1:00, while minimal values were observed at 5:00–9:00 in August in Lithuania. 

Discussion. It is possible to distinguish two regularities based on annual spore 

counts, comparing the results obtained from three aerobiological stations: 1) the 

lowest spore concentrations were in Vilnius, while higher in Šiauliai and Klaipėda 

aerobiological stations; 2) the interannual differences were smaller in Šiauliai than 

in the rest aerobiological stations – spore amount in Klaipėda in 2006 was more than 

twice as high as in 2005 and spore amount were 1.6 as high in 2006 as in Vilnius in 

2005. 

139

On the basis of 2005–2006 investigation, the highest total annual amount of 

Alternaria airborne spores occurred in Vilnius aerobiological station in 2006 (6223 

spores) and in Klaipėda in 2006 (9718 spores). Such abundance in Šiauliai maybe was 

caused by availability of host plants, because Alternaria is a common pathogen of 

cereals, vegetables, ornamentals, oilcrops, vegetables and fruits (Thomma, 2003). The 

investigations conducted in urban and rural environments in Poland confirm the preliminary 

hypothesis that daily concentrations, mean concentrations and seasonal sums 

of the studied fungal spores are lower in city air, than in areas with lower urbanisation 

level (Kaspryzk, Worek, 2006). Our sites according to town size and distance from 

its centre to the spore traps can be arranged by urbanisation level increase as follows: 

Klaipėda–Šiauliai–Vilnius. Easily can be noticed, that spore sums from all the investigation 

period according to sites are in decreasing order: 14 194–12 144–9 313. 

Alternaria airborne fungal spores occur in Lithuania throughout nearly the whole 

year. In February, March and April spores occur in air sporadically and in May, June 

concentrations of its conidia are usually low; the maximum falls in August, less – in 

July. According to Oliveira et al. (2005), the lowest concentrations of Alternaria spores 

were registered in winter and the highest concentrations were found in summer and 

early autumn in Porto (Portugal) in 2003. Spore concentrations were determined in 

summer in other places of Europe also: in Rzeszóv, Lublin and other cities of Poland 

(Kaspryzk et al., 2004; Konopińska, 2004; Stępalska et al., 1999; Stępalska, Wołek, 

2005), in Derby (England), Zagreb (Croatia) and other (Corden et al., 2003; Peternel 

et al., 2004). Corden et al. (2003) estimated that in two England towns Derby and 

Cardiff abundantly Alternaria spores are released in July, August and September, the 

main months for harvesting. Similar observations were made by Stępalska and Wołek 

(2005) in Cracow, Poland. The main months for release of Alternaria and other airborne 

fungal spores are July, August and September. These are the main months for 

combine harvesting and grass moving. 

Konopińska (2004) recorded in Liublin shorter time of Alternaria spore maximal 

value between 17:00–20:00, but longer minimal amounts value time: 0:00–7:00. 

Peternel et al. (2004) recorded in Zagreb minimal amounts of Alternaria spores on 

early morning, in 6:00. Munuera et al. (2001) observed that distribution of spore concentrations 

through the day was similar in 1993–1998 in Murcia, in Southern Spain: 

the maximal value of spore concentration was found in evening, at about 19:00, the 

lowest spore concentration was noted in early morning, at about 8:00. The same authors 

proposed that this late afternoon maximum could be related to a late release of 

spores from local sources or delay in sampling of spores released earlier and being 

transported to the sampler from rural areas (15–30 km away). 

Alternaria spore prevalence monitoring can be useful part of Integrated Disease 

Management programs. 

Conclusions. 1. The highest concentrations of Alternaria airborne spores in 

Lithuania were measured in the middle of spore season, i. e. in August. 51.4–70.3 % 

of annual spore amount was observed in this month. 

2. Depending on site and year, there were from 5 (Vilnius, 2005) to 28 (Klaipėda, 

2006) peak days with high concentration of Alternaria spores in the air. 

140

3. Hourly pattern of Alternaria spore concentration in August indicated the 

maximal value between 15:00 and 1:00, while minimal amounts were observed at 

5:00–9:00. 

Gauta 2009 06 30 


References 

1. Brazauskiene I., Petraitiene E. 2006. Epidemiological studies into Phoma lingam 

(telemorph Leptosphaeria maculans) infections in winter and spring oilseed 

rape. Agronomy Research, 4 (special issue): 137–140. 

2. Corden J. M., Millington W. M. 2001. The long-term trends and seasonal variation 

of the aeroallergen Alternaria in Derby, UK. Aerobiologia, 17(3/4): 127–136. 

3. Corden M. J., Millington M. W., Mullins J. 2003. Long-term trends and regional 

variation in the aeroallergen Alternaria in Cardiff and Derby UK – are differences 

in climate and cereal production having an effect Aerobiologia, 19(3/4): 

191–199. 

4. Käärik A., Keller J., Kiffer E., Perreau J., Resinger A. 1983. Atlas of airborne 

fungal spores in Europa. Berlin, Springer. 

5. Kasprzyk I., Rzepowska B., Wasylów M. 2004. Fungal spores in the atmosphere 

of Rzeszów (South-East Poland). Annals of Agricultural and Environmental 

Medicine, 11(2): 285–289. 

6. Kasprzyk I., Worek M. 2006. Airborne fungal spores in urban and rural environments 

in Poland. Aerobiologia, 22(3): 169–176. 

7. Konopińska A. 2004. Monitoring of Alternaria Ness and Cladosporium 

Link airborne spores in Lublin (Poland) in 2002. Annals of Agricultural and 

Environmental Medicine, 11(2): 347–351. 

8. Lacey W. E., West J. S. 2006. The air spore. Annual for catching and identifying 

airborne biological particles. Dordrecht, Springer. 

9. Lugauskas A., Paškevičius A., Repečkienė J. 2002. Pathogenic and toxic microorganisms 

in human environment. Vilnius, Aldorija. 

10. Lugauskas A., Šveistytė L., Ulevičius V. 2003. Concentration and species diversity 

of airborne fungi near busy streets in Lithuanian urban areas. Annals of 

Agricultural and Environmental Medicine, 10(2): 233–239. 

11. Mandrioli P., Comtois P., Levizzani V. 1998. Methods in aerobiology. Bologna: 

Pitagora Editrice. 

12. Mitakakis Z. T., Clift A., McGee A. P. 2001. The effect of local cropping activities 

and weather on the airborne concentration of allergenic Alternaria spores in 

rural Australia. Grana, 40(4/5): 230–239. 

13. Munuera M., Carrion J. S., Navarro C. 2001. Airborne Alternaria spores in SE 

Spain (1993–98) – occurrence patterns, relationship with weather variables and 

prediction models. Grana, 40(3): 111–118. 

141

14. Oliveira M., Ribeira H., Abreu I. 2005. Annual variation of fungal spores in 

atmosphere of Porto 2003. Annals of Agricultural and Environmental Medicine, 

12(2): 309–315. 

15. Peternel R., Čulig J., Hrga I. 2004. Atmospheric concentrations of Cladiosporium 

spp. and Alternaria spp. spores in Zagreb (Croatia) and effects of some meteorological 

factors. Annals of Agricultural and Environmental Medicine, 11(2): 

303–307. 

16. Petraitienė E. 2005. Occurrence and harmfulness of Alternaria blight (Alternaria 

spp.) in oilseed rape (Brassica napus var. oleifera) and turnip rape (Brassica 

rapa var. oleifera) and possibilities to reduce its damage: summary of doctoral 

dissertation. Kaunas. 

17. Stępalska D., Harmata K., Kasprzyk I., Myszkowska D., Stach A. 1999. 

Occurrence of airborne Cladosporium and Alternaria spores in Southern and 

Central Poland in 1995–1996. Aerobiologia, 15(1): 39–47. 

18. Stępalska D., Wołek J. 2005. Variation in fungal spore concentrations of selected 

taxa associated to weather conditions in Cracow, Poland, in 1997. Aerobiologia, 

21(1): 43–52. 

19. Sterling M., Rogers C., Levetin E. 1999. An evaluation of two methods used for 

microscopic analysis of airborne fungal spore concentrations from the Burkard 

Spore Trap. USA. Aerobiologia, 15(1): 9–18. 

20. Survilienė E., Valiuškaitė A. 2006. Carrot (Daucus sativus Röhl.) colonization 

by Alternaria spp. and effect of fungicide spray on their population. Ekologija, 

3: 54–59. 

21. Survilienė E., Šidlauskienė A., Vasinauskienė M., Zitikaitė I. 2004. Resistance of 

Brassica L. vegetables to phytopathogens. Horticulture and vegetable Growing 

23(1): 115–125, (in Lithuanian). 

22. Thomma J. H. P. B. 2003. Alternaria spp.: from general saprophyte to specific 

parasite. Molecular Plant Pathology, 4(4): 225–236. 


Ore plintančių Alternaria genties grybų sporų sklaidos ypatumai skirtingose 

Lietuvos vietovėse 

R. Mikaliūnaitė, M. Kazlauskas, L. Veriankaitė 

Santrauka 

Alternaria Ness. genties grybų sporų paplitimas ore tyrimo metais, sezono metu 

bei pasiskirstymas paros metu 2005–2006 metais tirtas trijose Lietuvos urbanistinėse 

vietovėse – Klaipėdoje, Šiauliuose ir Vilniuje. Nustatyti grybų sporų kiekiai per metus, 

sezono metu bei pasiskirstymas paros metu. Sporų fiksavimui naudota Hirsto tipo 

sporų gaudyklė. Grybų sporos buvo identifikuojamos ir skaičiuojamos 12 vertikalių 

juostų metodu. Grybų sporų sezono trukmė apskaičiuota taikant 90 % metodą. 

142

Tyrimo metu sporų sezono trukmė buvo nuo 62 dienų Vilniuje 2005 metais iki 97 dienų 

Klaipėdoje 2006 metais. Gausiausi Alternaria sporų kiekiai buvo nustatyti vasarą, o 

mažiausi – žiemą bei pavasarį. Visose aerobiologinėse stotelėse pavienės sporos nustatytos 

vasario, kovo ir balandžio mėnesiais. Bendras Alternaria sporų kiekis per metus įvairavo 

nuo 3 090 sporų Vilniaus aerobiologinėje stotelėje 2005 metais iki 9 718 sporų Klaipėdoje 

2006 metais. Sporifikacijos sezono pikai nustatyti rugpjūčio mėnesį visose tirtose vietovėse 

ir įvairavo nuo 121 iki 707 sporų m -3 . Šį mėnesį buvo nustatyta 51,4–70,3 % sporų nuo 

metinio sporų kiekio. Paros metu gausiausios Alternaria sporų koncentracijos rugpjūčio 

mėnesį buvo nustatytos tarp 15:00 ir 1:00 valandos, mažiausios – 5:00–9:00 valandomis. 

Reikšminiai žodžiai: Alternaria, grybų sporos, sezono trukmė, sporų koncentracija. 

143




Water as the source of Phytophthora spp. 

pathogens for horticultural plants 

Leszek B. Orlikowski, Magdalena Ptaszek, Aleksandra Trzewik, 

Teresa Orlikowska 

Res. Institute of Pomology & Floriculture, Pomologiczna 18, 96-100 Skierniewice, 

Poland, e-mail lorlikow@insad.pl 

Using rhododendron leaf blades, Phytophthora spp. was recovered from 4 rivers, 3 

reservoirs and 3 canals located in different parts of Poland. Independently of water sources 

location, P. citricola was found in rivers, reservoirs and canals. Detection period and different 

sources of water had no big influence on Phytophthora spp. population density. Occurrence 

of P. cactorum, P. cambivora and P. cinnamomi in sampling water was influenced by presence 

of potential host plants near river and in nurseries. Under conditions favourable to the 

development of Phytophthora spp., P. citricola dispersed with sprinkling water in 2 hardy 

ornamental nursery stocks, caused shoot and tip blight of boxwood and thuja. 

Key words: density, dispersal, harmfulness, leaf bait, Phytophthora, recovery, water. 

Introduction. Most of Phytophthora species are known as economically important 

plant pathogens causing mainly root and stem base rot, leaf spots, fruit necrosis 

and tip blight. Development of disease symptoms is favoured by wet soil or 

substratum conditions and temperature above 20 °C resulting rapid dispersal of that 

group of pathogens achieved by zoospores. The most actual example of zoospores 

spread in water is P. alni known since 1999 on Alnus glutinosa firstly in England and 

during the next few years in many countries of Europe (Cech, 2004; Gibbs et al., 

Orlikowski et al., 2003). The occurrence of Phytophthora species in rivers, streams, 

canals and water reservoirs is well documented. At least 20 Phytophthora spp. were 

detected in water mainly in the USA and Europe including the most known species 

like P. cactorum, P. cambivora, P. capsici, P. cinnamomi, P. citricola, P. citrophthora, 

P. cryptogea, P. drechsleri, P. lateralis, P. megasperma, P. nocotianae, var. nicotianae, 

P. ramorum, P. syringae (Fergusson and Jeffers, 1999; Orlikowski, 2006; Themann 

et al., 2002). Hong and Moorman (2005) characterized Phytophthora spp. as 

significant crop health destructor, which increased greatly and will remain as agriculture 

problem. The authors concluded that contaminated water irrigation is a primary, 

if not a sole, source of inoculum for Phytophthora diseases of numerous nursery, 

145

fruit, and vegetable crops. This conclusion is the key for studying of Phytophthora 

detection methods, population densities in relation to water sources and weather conditions 

and economic thresholds. 

The aim of this study is to present some results connected with the occurrence 

of Phytophthora species in Polish rivers, canals, water reservoirs and harmfulness of 

P. citricola to some ornamental nursery plants. 

Object, methods and conditions. S o u r c e s o f w a t e r. Phytophthora spp. 

was detected in 4 rivers, 3 water canals and 3 reservoirs. Rivers were situated in different 

parts of Poland. One of them (Korabiewka) is flowing through forest area, the 

second one (Rawka) – through agricultural area and partly forest, whereas the next 

two – through horticultural area with hardy nursery stocks and greenhouse farms 

(Kurówka and Ner). Water canals and reservoirs are situated in 3 hardy ornamental 

nursery stocks in different parts of the country. 

Recovery of Phytophthora f r o m w a t e r. Rhododendron leaflets 

baits cv. ‘Nova Zembla’ were used. Baits containing at least 8 leaves secured with about 

3 m pieces of string were held in water about 2–3 m from benches. After 4–7 days of 

incubation, in relation to part of the year, baits were removed from water, washed under 

tap water and number of necrotic spots on each leaf blades was counted. Chosen leaves 

were rinsed in distilled water, blotted dry, sterilized over a burned flame and about 

2–5 mm in diameter pieces of leaves with brown or dark-brown spots were placed on 

PDA medium and incubated at 25 °C in the dark. After 24–48 h small parts of possible 

Phytophthora colonies were transferred to PDA slants. After segregation and cleaning 

of chosen isolates they were identified with species on the base of morphology and 

using molecular methods (Trzewik et al., 2006). 

Harmfulness of Phytophthora cotricola t o b o x w o o d a n d 

t h u j a. Observations were done in two hardy nursery stocks. In both of them plants 

were regularly sprinkled with water taken from reservoirs situated in the lowest part of 

nurseries. Part of water was taken from wells but during summer time also from local 

rivers. Additionally surplus of water from nurseries was flowing to both reservoirs. 

First symptoms of leaf yellowing were observed on boxwood (Buxus sempervirens) in 

the beginning of June, whereas dying of thuja (Thuja occidentalis Fastigiata) tips – in 

the second decade of July. Within one week (thuja) and four months (boxwood) development 

of diseases were observed. Experimental design was completely randomized 

with 4 replications and 200 plants in each rep. 

Results. Recovery of Phytophthora s p p . f r o m w a t e r. Results 

with baiting of Phytophthora from rivers indicated significantly higher population 

number of that group of organisms in August than in June only. In Rawka significantly 

more necrotic spots on baiting leaves were observed in June and Phytophthora 

population density was about twice higher than in other rivers (Fig. 1, A). The river 

runs mainly through forest and agricultural area, so probability of water refuse by 

chemical residues is much lower than in Ner and Kurówka flowing through horticultural 

localities (Fig. 1, A). 

146

Fig. 1. Occurence of Phytophthora spp. in rivers (A), water reservoirs (B) and 

nursery canals (C); number of spots on rhododendron leaf blade 

1 pav. Phytophtora spp. paplitimas upėse (A), medelynų kanaluose (B) ir 

vandens rezervuaruose (C); ligos požymiai ant rododendrų lapalakščių 

147

Analysis of Phytophthora spp. detection from water reservoirs and nursery canals 

showed the lack of differences in population densities as well as between baiting period 

(Fig. 1, B, C). Four Phytophthora species were detected from sampling water sources. 

P. citricola was detected from all sampling water in June and August. P. cinnamomi 

was found only in Kurówka river, P. cactorum – in Ner, meanwhile P. cambivora – in 

Rawka. 

Harmfulness of Phytophthora citricola t o b o x w o o d a n d 

t h u j a. The first yellowing of individual boxwood shoots was observed in the middle 

of June on about 1.5 of plants (Fig. 2). During the next 15 weeks disease symptoms 

were noticed on at least 10 % of plants. Besides yellowing, on about 5 % of plants 

most of shoots showed dark-brown discoloration (Fig. 2). 

Fig. 2. Development of Phytophthora shoot rot (P. citricola) of 

Buxus sempervirens in hardy ornamental nursery stock 

2 pav. Buxus sempervirens šaknų puvinio Phytophtora (P. citricola) 

vystymasis atspariame dekoratyvinių medžių medelyne 

On tops of T. occidentalis disease symptoms were noticed after 2 days at relative 

air humidity above 90 %. Single tips changed colour to yellow-brown and dark-brown 

and the disease developed very quickly (Fig. 3). First observation showed about 7 % 

of diseased plants and within 8 weeks symptoms spread on about 40 % of plants (Fig. 

3). 

148

Fig. 3. Spread of Phytophthora tip blight (P. citricola) on 

Thuja occidentalis Fastigiata 

3 pav. Phytophthora P. citricola galų rūdžių paplitimas tujose 

Thuja occidentalis Fastigiata 

Discussion. Phytophthora species were isolated from rivers, water reservoirs 

and nursery canals all the year. Results are given for recovery of them in June and 

August because of available temperature for Phytophthora development, but also – 

the possibility of chemical residues transport from nurseries to water reservoirs, canals 

and rivers. In Themann and et al. (2002) studies detection rates of Phytophthora 

spp. from nursery drains were higher in most cases than from reservoirs. The authors 

found also great variation between nurseries and sampling sites. Studies of 

Orlikowski et al. (2007) indicated that P. citricola is the dominant water species. 

Occurrence of P. cinnamomi in Kurówka river was probably connected with running 

of surplus water from hardy nursery stocks where this species was causal agent of 

root and stem rot of Chamaecyparis lawsoniana, Pinus mugho var. pumilo and other 

coniferous plants (Orlikowski et al., 1995). P. cambivora occurs in forests, especially 

on European beech (Orlikowski et al., 2006), whereas P. cactorum – on ericaceous 

plants located near Ner river. 

The thuja is the best example indicating on very fast spreading of Phytophthora 

tip blight with water and expanding of the disease on young plant tips and in nurseries 

(Orlikowski, 2006; Orlikowski et al., 2004). Our study showed an important role 

of water in the dispersal of Phytophthora diseases not only inside nurseries but also 

around the country. 

Conclusions. 1. Rhododendron leaf baits were an excellent medium for recovery 

of Phytophthora species from different sources of water. 

2. Location of rivers and surrounding area and recovery period had no great 

influence on population densities of Phytophthora spp. in water. 

149

3. Similar spots number on rhododendron leaf baits were noticed independently 

of localization of water reservoirs and canals in the country. 

4. From rivers, reservoirs and water canals Phytophthora citricola, P. cinnamomi, 

P. cactorum and P. cambivora were recovered with domination of the first species. 

5. Under conditions favourable to the development of Phytophthora spp., P. citricola 

dispersal on plants with sprinkling water may cause high losses in the production 

of some ornamental coniferous and deciduous plants. 

Gauta 2009 07 03 


References 

1. Cech Th. L. 2004. Development and spread of Phytophthora disease of alders in 

Austria. Materials of the 3rd IUFRO Working Party “Phytophthora in forest and 

natural ecosystems”, Freising, Germany 2004.09. 31: 11–17. 

2. Gibbs J. N., Lipscombe M. A., Peace A. J. 1999 The compact of Phytophthora 

disease on riparian populations of common alder (Alnus glutinosa) in southern 

Britain. Eur. J. For. Pathol., 29: 39–50. 

3. Fergusson A. J., Jeffers S. N. 1999. Detecting multiple species of Phytophthora in 

container mixes from ornamental crop nurseries. Plant Dis., 83: 1 129–1 136. 

4. Hong C. X., Moorman G. W. 2005. Plant pathogens in irrigation water: challenges 

and opportunities. Critical Rev. in Pl. Sci., 24: 189–208. 

5. Orlikowski L. B. 2006. Relationship between source of water used for sprinkling 

and occurrence of Phytophthora shoot rot and tip blight in container-ornamental 

nurseries. J. Pl. Prot. Res., 46: 163–168. 

6. Orlikowski L. B., Gabarkiewicz R., Skrzypczak Cz. 1995. Phytophthora species 

in Polish ornamental nurseries: I. Isolation and identification of Phytophthora 

species. Phytopathol., 9: 73–79. 

7. Orlikowski L. B., Szkuta G., Sroczyński M. 2004. First notice of Phytophthora 

tip blight of Calluna vulgaris. Phytopathol Pol., 31: 67–71. 

8. Orlikowski L. B., Trzewik A., Orlikowska T. 2007. Water as potential source of 

Phytophthora citricola. J. Plant prot. Res., 47: 125–132. 

9. Themann K., Werres S., Luttmann R., Diener H.-A. 2002. Observations of 

Phytophthora spp. in water recirculation systems in commercial hardy ornamental 

nursery stocks. Eur. J. Plant Pathol., 108: 337–343. 

10. Trzewik A., Wiejacha K., Orlikowski L. B., Orlikowska T., 2006. Identification 

of five Phytophthora species, causal agents of diseases of nursery perennials, 

trees and shrubs on the base of DNA markers amplified with non-specific primers. 

Phytopathol. Pol, 41: 27–37. 

150


Vanduo kaip sodo augalų ligų sukėlėjo Phytophthora spp. šaltinis 

L. B. Orlikowski, M. Ptaszek, A. Trzewik, T. Orlikowska 

Santrauka 

Panaudojant rododendrų lapalakščius, Phytophthora spp. buvo išgauta iš 4 upių, 3 vandens 

rezervuarų ir 3 kanalų, esančių skirtingose Lenkijos vietose. Nepriklausomai nuo vandens telkinio 

vietovės, P. citricola rastas upėse, vandens rezervuaruose ir kanaluose. Aptikimo laikotarpis 

ir skirtingi vandens šaltiniai neturėjo didelės įtakos Phytophthora spp. populiacijos tankumui. 

Tam, kad vandens mėginiuose buvo rasta P. cactorum, P. cambivora ir P. cinnamomi, turėjo 

įtakos potencialiai užkrėsti augalai netoli upės ir medelynuose. Esant Phytophthora spp. vystymuisi 

palankioms sąlygoms, P. citricola paplitęs su laistomu vandeniu dviejuose atspariuose 

dekoratyvinių medžių medelynuose, sukėlė buksmedžių ir tujų šaknų bei viršūnių rūdis. 

Reikšminiai žodžiai: išgavimas, kenksmingumas, lapų diskų metodas, paplitimas, 

Phytophthora, tankumas, vanduo. 

151




The toxicity of Neem to the snail 

Arianta arbustorum 

Angela Ploomi*, Katrin Jõgar, Luule Metspalu, Külli Hiiesaar, 

Liina Loorits, Ivar Sibul, Irja Kivimägi, Anne Luik 

Estonian University of Life Sciences, 1 Kreutzwaldi St., 51014 Tartu, Estonia, 

e-mail angela.ploomi@emu.ee 

Herbivorous land snail Arianta arbustorum Linnaeus, 1758 (Gastropoda, Pulmonata, 

Helicidae) has become considerable pest that occurs throughout Central, Eastern and Northern 

Europe. Commercially available neem formulation NeemAzal-T/S (containing 1 % azadirachtin, 

Trifolio-M GmbH, Germany) was tested on experimental white cabbage field of the 

Estonian University of Life Sciences against cabbage pests, including the snail A. arbustorum. 

The formulation was diluted with water and treated in concentrations 0.03 % and 0.3 % (solution/spraying 

and watering) with weekly intervals. After that the number of snails started 

to grow and the highest population peak was in the middle of September. All the tested neem 

concentrations affected the number of snails on the cabbage and were significantly different 

from control variant. There was no significant difference between treated variants. Both, 

spraying and watering with NeemAzal-T/S, affected A. arbustorum as effective repellent. It 

can be concluded that all neem treatments could control the snail A. arbustorum. 

Key words: Arianta arbustorum L., NeemAzal-T/S, white cabbage. 

Introduction. A lot of terrestrial gastropod species are important pests in agricultural 

and horticultural crops (Godan, 1983). The commonest and widespread hermaphroditic 

land snail Arianta arbustorum Linnaeus, 1758 (Gastropoda, Pulmonata, 

Helicidae) occurs throughout Central, Eastern and Northern Europe (Terhivuo, 1978; 

Kerney et al., 1983). A. arbustorum is an omnivorous snail that includes green plants, 

decaying plant material, and fungi in its diet (Hägele, 1992). The most commonly 

used chemical control is in the form of slug pellets, containing the active ingredient 

metaldehyde. Metaldehyde works by disrupting the gastric organs. Molluscicides 

may also contain a desiccant preventing mollusks from producing the mucous 

essential for survival. Pellets remain attractive to snails and slugs for several days to 

prolong the length of time they are available for detection (Pesticide Safety Directive, 

2000). It is tempting to use chemical control in our gardens but care must always be 

taken as all pesticides are toxic to living organisms and the potential hazards are very 

real. With the increasing popularity of organic farming in the country, there is a need 

to use environment-friendly biopesticides and other bio-based tools to overcome the 

153

problem of insect pests. Plants may be an alternative to the currently used insect control 

agents, because they virtually are a rich source of bioactive organic chemicals 

(Godfrey, 1995; Isman, 1997). Phytophagous land snails in greenhouses and horticulture 

were killed by neem preparations (West, Mordue, 1992). Biopesticide NeemAzal-T/S 

(Trifolio-M GmbH, Germany) is registered since 2000 in Germany (Isman, 

2004) and since 2001 in Estonia. An extract “NeemAzal” obtained from seed kernels 

of the Neem tree Azadirachta indica A. Juss and its formulation contains about 54 % 

of azadirachtins. NeemAzal-T/S is a formulation of NeemAzal containing 1 % w/w 

of azadirachtin A. The results of studies of possible environmental impacts indicate 

an extremely low risk to aquatic organisms, microorganisms, earthworms, beneficial 

insects, honeybees, birds, etc. This is true especially in view of the low concentrations 

of azadirachtin A, which are required for sufficient control of pests (Kleeberg, 

2004). Aim of the present study was to test different concentrations and treatment 

ways of neem formulation NeemAzal-T/S on white cabbage field against the herbivorous 

land snail A. arbustorum. 

Object, methods and conditions. Field experiments were done to test the 

efficacy of the biopesticide NeemAzal-T/S (1 % azadirachtin, Trifolio-M GmbH, 

Germany) against the cabbage pests, including the snail A. arbustorum. The present 

experiment was carried out in the experimental garden of the Estonian University of 

Life Sciences in 2006. White cabbage (Brassica oleracea L. var. capitata f. alba) 

plants were grown from seed, kept in glasshouse until they reached the 3 true leaf 

stage. In mid-May the plants were replanted to the experimental field. Each variant 

consisted of 9 plants per plot (three rows of three plants spaced at 70 cm intervals). 

All variants had three replications. Different concentrations of NeemAzal-T/S were 

used – 0.03 % and 0.3 % for spraying and 0.3 % concentration of NeemAzal-T/S for 

watering of testing plots. Individuals of A. arbustorum on all the plots were sampled 

at 7 day intervals from 4 July to 12 September. To avoid repeated counting the snails 

were removed from plants by hand picking. The first spraying and watering with 

NeemAzal-T/S was made after the first counting on 4th of July. Treatments were 

applied at weekly intervals during the entire observing period. Tests were performed 

using the statistical package StatSoft ver. 7, Inc. / USA. Data have been presented as 

mean ± standard error. Statistical comparisons were performed with repeated-measures 

by the Fisher LSD test. All means were considered significantly different at the 

p < 0.05 level. 

Results. The snail A. arbustorum appeared on the white cabbage in the beginning 

of August whereupon the number of snails started to grow. The highest population 

peak was in the middle of September (Fig. 1). 

All the tested neem concentrations affected the snail and were significantly different 

from control variant. A comparison of treated variants with control revealed 

significant differences in the number of A. arbustorum individuals (ANOVA F(3.128) = 

8.75; df = 3; p < 0.05: LSD test p < 0.05). There was found no significant difference 

among treated variants’ snail numbers (LSD test, p > 0.05) (Fig. 2). 

154

Fig. 1. Seasonal dynamics of land snail Arianta arbustorum on differently 

NeemAzal-T/S treated variants (mean ± SE) 

1 pav. Margųjų medsraigių Arianta arbustorum sezoninis gausumo kitimas 

skirtingai nimazaliu apdorotuose variantuose (vidurkis ± SE) 

Fig. 2. Mean number of individuals of land snail Arianta arbustorum on neem treated 

variants. Columns with different letters are significantly different (Fisher LSD test p < 0.05) 

2 pav. Vidutinis margųjų medsraigių Arianta arbustorum skaičius neem purkštuose 

variantuose. Stulpeliai pažymėti skirtingomis raidėmis patikimai skiriasi (Fišerio testas R 05 

) 

155

These data suggest that A. arbustorum was sensitive to NeemAzal-T/S because 

the number of snails remained relatively low on treated variants. The comparison of 

the differently treated variants revealed that the land snail A. arbustorum selected 

least the plots sprayed with 0.3 % neem formulation (7 %), followed by sprayed with 

0.03 % neem (11 %) as the site for feeding and the third choice was watered variant 

0.3 % neem (17 %). 

Discussion. Mollusc herbivory is generally considered to be of ecological importance 

only during seedling period (Hanley et al., 1995 a, b). However, in suitable 

mollusk habitats, mollusk biomass is high, and they may well be a major basal component 

of the food chain. Consequently, mollusk in these habitats should not only 

affect seedlings but also exert some pressure on fully-grown plants (Hägele et al., 

1998). Damage is caused by gastropods due both to feeding and to contamination 

of the harvested plants with their bodies, eggs, faeces or slime, leading to deterioration 

in the quality of the harvest and financial loss (South, 1992). The compounds 

from neem A. indica have a number of properties useful for pest management. These 

include repellence, feeding and oviposition deterrence, insect growth regulator activity, 

low mammalian toxicity and low persistence in the environment (Schmutterer, 

1990; Koul, 1992). It is evident from our results that the NeemAzal-T/S we tested 

had a repellent effect on the snail A. arbustorum. Some test results prove the neem 

oil molluscicidal activity. Neem oil significantly reduced fecundity, egg viability, and 

survival of the giant African snail Achatina fulica (Rao, Singh, 2000). Neem oil resulted 

in extension of the nymphal period and dose-dependent mortality in some 

homopterous insects (Schmutterer, 1990). Singh and Singh (2000) while studying the 

effect of Allium sativum, A. indica and Zingiber officinale on the reproduction of the 

snail Lymnaea acuminata reported that their active molluscicidal constituents allicin, 

azadirachtin, and [6]-gingerol (Singh, Singh, 1995; Singh et al., 1996; 1997) caused 

a significant reduction in the fecundity, egg viability, and survival of young snails. 

Treatment of 60 % of LC50/24 h of allicin and [6]-gingerol eliminated fecundity in 

snail L. acuminata. 

Conclusion. In our research both spraying and watering with NeemAzal-T/S 

affected A. arbustorum as effective repellent. It can be finally concluded that all neem 

treatments could affect the snail A. arbustorum. 

Acknowledgements. This research was supported by the Estonian Science 

Foundation (grants no 7130, 6722 and 6781), and Estonian Ministry of Education 

and Research targeted financing projects no. SF170057s09, SF0170014s08 and 

SF0170021s08. 

Gauta 2009 06 30 


156

References 

1. Godan D. 1983. Pest slugs and snails: biology and control. Springer-Verlag. 

Berlin. 

2. Godfrey C. R. A. (ed.). 1995. Agrochemicals from Natural Products. New York. 

3. Hanley M. E., Fenner M., Edwards P. J. 1995 b. An experimental field study of 

the effects of mollusk grazing on seedling recruitment and survival in grassland. 

Journal of Ecology, 83: 621–627. 

4. Hanley M. E., Fenner M., Edwards P. J. 1995 a. The effect of seedling age on the 

likelihood of herbivory by the slug Deroceras reticulatum. Functional Ecology, 

9: 754–759. 

5. Hägele B. 1992. Saisonale Änderungen der Nahrungswahl des generalistischen 

herbivoren Arianta arbustorum (L.) in verschiedenen, von Pflanzen aus 

der Tribus Senecioneae dominierten Habitaten. Masters thesis. University of 

Tübingen, Tübingen. 

6. Hägele B. F., Wildi E., Harmatha J., Pavlík M., Rowell-Rahier M. 1998. Longterm 

effects on food choice of land snail Arianta arbustorum mediated by petasin 

and furanopetasin, two sequiterpenes from Petasites hybridus. Journal of 

Chemical Ecology, 24(11): 1 733–1 743. 

7. Isman M. B. 1997. Neem and other botanical insecticides: barriers to commercialization. 

Phytoparasitica, 25(4): 339–344. 

8. Isman M. B. 2004. Factors limiting commercial success of neem insecticides in 

North America and Western Europe. In: O. Koul, S. Wahab (eds.). Neem: Today 

and in the New Millenium. Kluwer Academic Publishers. Netherland, 33–41. 

9. Kerney M. P., Cameron R. A. D., Jungbluth J. H. 1983. Die landschnecken Nordund 

Mitteleuropas. Paul Parey. Hamburg und Berlin. 

10. Kleeberg H. 2004. Neem based products: registration requirements, regulatory 

processes and global implications. In: O. Koul, S. Wahab (eds.). Neem: 

Today and in the New Millenium. Kluwer Academic Publishers. Netherland, 

109–123. 

11. Koul O. 1992. Neem allelochemicals and insect control. In: R. S. H. J. Rizvi, 

V. J. Rizvi (eds.). Allelopathy: basic and applied aspects. Chapman & Hall Ltd. 

London. 389–412. 

12. Pesticide Safety Directive, Efficacy Guideline 510. Testing of Molluscicide 

Products, 23 March 2000, PSD, Mallard House, Kings Pool, York YO1 7PX 

http://193.133.84.30/ 

13. Rao I. G., Singh D. K. 2000. Effect of single and binary combinations of plantderived 

molluscicides on reproduction and survival of the snail Achatina fulica. 

Archives of Environmental Contamination ant Toxicology, 39: 486–493. 

14. Schmutterer H. 1990. Properties and potential of natural pesticides from neem 

tree, Azadirachta indica. Annual Review of Entomology, 35: 271–297. 

15. Singh K., Singh D. K. 1995. Molluscicidal activity of Azadirachta indica (neem) 

on biochemical parameters in the ovotestis of Lymnaea acuminata. Malaysian 

Applied Biology, 24: 7–11. 

157

16. Singh K., Singh D. K. 2000. Effect of different combinations of MGK-264 or piperonyl 

butoxide with plant-derived molluscicides on snail reproduction. Arch. 

Environ. Contam. Toxicol, 38: 182–190. 

17. Singh K., Singh A., Singh D. K. 1996. Molluscicidal activity of neem (Azadirachta 

indica). J. Ethnopharmacol, 52: 35–40. 

18. Singh S., Singh V. K., Singh D. K. 1997. Molluscicidal effect of some common 

spice plants. Biol. Agricul. Horticul., 14: 237–249. 

19. South A. 1992. Terrestrial Slugs. Biology, Ecology, Control. London: Chapman 

& Hall Ltd. 

20. Terhivuo J. 1978. Growth, reproduction and hibernation of Arianta arbustorum 

(L.) (Gastropoda, Helicidae) in southern Finland. Ann. Zool. Fennici, 15: 

8–16. 

21. West A. J., Mordue (Luntz) A. J. 1992. The influence of azadiractin on the feeding 

behaviour of cereal aphids and slugs. Entomol. Exp. Appl., 62: 75–79. 


Neem toksiškumas medsraigei Arianta arbustorum 

A. Ploomi, K. Jõgar, L. Metspalu, K. Hiiesaar, L. Loorits, 

I. Sibul, I. Kivimägi, A. Luik 

Santrauka 

Margosios medsraigės Arianta arbustorum Linnaeus, 1758 (Gastropoda, Pulmonata, 

Helicidae) tampa žalingais kenkėjais. Jų randama centrinėje-rytinėje ir šiaurinėje Europoje. 

Pramoniniu būdu gaminama neem formuluotė nimazalis-T/S (turintis 1 % azadirachtino, 

Trifolio-M GmbH, Germany) buvo bandomas Estijos gamtos mokslų universiteto eksperimentiniuose 

baltagūžių kopūstų laukuose kovojant su kopūstų kenkėjais ir sraigėmis A. arbustorum. 

Preparatas buvo atskiestas vandeniu ir naudotas 0,03 % ir 0,3 % koncentracijų (tirpalas purkšti 

arba laistyti) kas savaitę. A. arbustorum pasirodo rugpjūčio pradžioje. Po to sraigių gausėja, ir 

daugiausia jų būna rugsėjo viduryje. Visos bandytos neem koncentracijos veikė sraiges ant kopūstų 

ir patikimai skyrėsi nuo kontrolinio varianto. Tarp apdorotų variantų patikimų skirtumų nebuvo. Ir 

laistymas, ir purškimas nimazaliu-T/S buvo veiksmingas A. arbustorum repelentas. Galima daryti 

išvadą, kad ir purškimas, ir laistymas nimazaliu gali reguliuoti sraigių A. arbustorum skaičių. 

Reikšminiai žodžiai: Arianta arbustorum L., baltagūžiai kopūstai, nimazalis. 

158




New host plants for development of 

Phytophthora cryptogea in Poland 

Magdalena Ptaszek, Leszek B. Orlikowski, 

Czesław Skrzypczak 

Res. Institute of Pomology & Floriculture, Pomologiczna 18, 96–100 Skierniewice, 

Poland, e-mail magdalena.ptaszek@insad.pl 

Pathogenicity of nine isolates of Phytophthora cryptogea from different host plants was 

tested. In the laboratory trials, all cultures (except S. arachnoideum) colonized leaf blades 

and stem parts of 3 alstroemeria cultivars, but isolates from Anthurium andreanum, Aquilegia 

discolor and Alstroemeria × hybrida were the most pathogenic. In the tests with columbine 

the quickest spread of necrosis was observed on leaves inoculated with P. cryptogea from 

A. discolor and Gerbera jamesonii, whereas the slowest when isolates from Sempervivum 

arachnoideum was used. Inoculation of S. arachnoideum with eight isolates of P. cryptogea 

caused necrosis of leaves, but isolates from A. discolor, Campanula persicifolia and S. arachnoideum 

were the most pathogenic. 

Key words: colonization, host plants, isolation, occurrence, pathogenicity, Phytophthora 

cryptogea. 

Introduction. The studies of Phytophthora spp. conducted in Poland during 

the last 10 years showed the occurrence of 17 species and increasing number of new 

host plants for these pathogens. Phytophthora species were mainly observed on ericaceous 

and coniferous plants. P. cinnamomi and P. citricola were the casual agent 

of root rot and stem rot of rhododendron, heather, cypress, yew and pine plants (Orlikowski 

1996, Orlikowski, Szkuta 2002b, 2003, Orlikowski et al. 2004). In the first 

decade of XXI century P. cryptogea was the most often recorded on diseased plants 

both in greenhouses and hardy ornamental nursery stocks. According to Erwin and 

Ribeiro (1996), P. cryptogea is one of the most dangerous pathogen, infected about 

150 plant species belonged to 23 botanical family. In Poland the species was known 

for at least 40 years as the casual agent of foot rot of gerbera (Orlikowski 1967/77). 

The objective of this work was to establish the occurrence of P. cryptogea on 

Aquilegia discolor, Alstroemeria × hybrida, Sempervivum arachnoideum and its 

pathogenicity toward 3 plant species. 

159

Object, methods and conditions. Isolation of Phytophthora cryptogea 

f r o m t h e d i s e a s e d p l a n t p a r t s. Plants with diseased symptoms 

were taken in greenhouse plantations and hardy nursery stocks from June to September 

2006–2008. Diseased plants or their parts were collected in plastic bags and transferred 

to the laboratory. The same or next day the fragments of plants were washed 

under tap water, rinsed in distilled water and after blotting dried and sterilized over 

a burner flame, 5 mm diameter pieces of tissues were put on potato-dextrose agar 

(Orlikowski and Szkuta, 2002). Colonies grown around inocula were transferred into 

PDA slants. After 7 days cultures obtained were grouped and selected isolates were 

identified to genera and species on the base of their morphology. The results were 

confirmed using PCR method with species-specific primers for P. cryptogea CRYF2/ 

CRYR2 (Boersma et al., 2000). 

Colonisation of plant parts by Phytophthora cryptogea. 

Phytophthora cultures. Isolates of P. cryptogea from Anthurium andreanum, Aquilegia 

discolor, Alstroemeria × hybrida, Campanula persicifolia, Forsythia intermedia, 

Gerbera jamesonii, Lycopersicon esculentum, Saxifraga arendsii and Sempervivum 

arachnoideum were used for inoculation of plant parts of alstroemeria, columbine 

and sempervivum. Stock cultures were grown on PDA at 24 °C in the dark. For plant 

parts inoculation five mm diameter inocula were taken from the edge of 7-day-old 

cultures. 

I n o c u l a t i o n o f p l a n t p a r t s. Leaf blades and about 8 cm long stem 

parts of tested plants were placed in a tray covered with moist blotting paper and 

plastic net. The central parts of leaves and the bases of stem pieces were inoculated 

with P. cryptogea inoculum. Trays were covered with plastic foil. After 4–5 days of 

incubation the size (diameter and length) of necrosis was measured. Experimental 

design was completely randomized with 4 replications and 10 leaf blades and stem 

parts in each replication. The trials were repeated at least twice. 

Results. Results of laboratory trials with colonization of alstroemeria, columbine 

and houseleek by P. cryptogea from 9 different host plants showed significant 

differences in pathogenicity of tested isolates toward all plant species and cultivars. 

In trials with alstroemeria plants all isolates (except S. arachnoideum) colonized stem 

parts and leaf blades (results not shown) of three cultivars. After 4-day incubation 

the quickest spread of necrosis was observed on alstroemeria inoculated with culture 

from A. discolor and A. × hybrida, whereas the slowest – on stem parts treated 

with isolates from S. arendsii and S. arachnoideum (Table). On columbine significant 

differences of disease spread were especially noticed after 5-day incubation. 

The quickest spread of necrotic spots was observed when isolates from G. jamesonii 

and A. discolor were used for inoculation of leaves, whereas the slowest with culture 

from S. arachnoideum (Fig. 1). 

Inoculation of S. arachnoideum with isolates of P. cryptogea from greenhouse 

plants as well as from field plants showed that all of them caused necrosis of leaves, 

but isolates from A. discolor, C. persicifolia and S. arachnoideum were the most 

pathogenic (Fig. 2). 

160

Table. Relationship between different isolates of Phytophthora cryptogea and 

the development of necrosis on 3 cultivars of Alstroemeria x hybrida stem parts: 

length of necrosis (mm) 4 days after inoculation 

Lentelė. Ryšys tarp skirtingų Phytophthora cryptogea izoliatų ir nekrozės vystymosi 

ant 3 Alstroemeria x hybrida veislių stiebų: nekrozės ilgis (mm), praėjus 4 dienoms po 

inokuliacijos 

Source of P. cryptogea 

P. cryptogea augalas šeimininkas 

Alstroemeria cultivars 

Alstroemeria veislės 

‘Lorena’ ‘Simeona’ ‘Tiara’ 

Anthurium andreanum 8.4 b 6.1 b 4.5 c 

Aquilegia discolor 9.6 b 11.0 e 5.7 d 

Alstroemeria × hybrida 12.5 c 8.5 d 6.2 e 

Gerbera jamesonii 6.1 a 7.4 c 3.0 b 

Saxifraga arendsii 6.3 a 4.4 a 3.0 b 

Sempervivum arachnoideum 8.6 b 4.7 a 0 a 

Note: Means followed by the same letter do not differ significantly (p = 0.05) according to 


Pastaba: Ta pačia raide pažymėtos reikšmės pagal Dunkano kriterijų patikimai nesiskiria 

(p = 0,05) 

Fig. 1. Spread of necrosis on Aquilegia discolor leaves 3 and 5 days 

after plant inoculation by isolates of P. cryptogea from 5 different host plants. 

Note: Means followed by the same letter do not differ significantly 

(p = 0.05) according to Duncan’s multiple range test 

1 pav. Nekrozės plitimas ant Aquilegia discolor lapų praėjus 3 ir 5 dienoms po 

augalo inokuliacijos P. cryptogea izoliatais iš 5 skirtingų augalų. 

Pastaba: Ta pačia raide pažymėtos reikšmės pagal Dunkano kriterijų patikimai nesiskiria (p = 0,05) 

161

Fig. 2. Spread of necrosis on S. arachnoideum leaves 4 days after plant 

inoculation with isolates of P. cryptogea from 8 different host plants. 

Note: Means followed by the same letter do not differ significantly 

(p = 0.05) according to Duncan’s multiple range test 

2 pav. Nekrozės plitimas ant S. arachnoideum lapų praėjus 4 dienoms po 

augalo inokuliacijos P. cryptogea izoliatais iš 8 skirtingų augalų. 

Pastaba: Ta pačia raide pažymėtos reikšmės pagal Dunkano kriterijų patikimai nesiskiria (p = 0,05) 

Discussion. During the last two years P. cryptogea was isolated from the most of 

analysed and diseased plants with stem/leaf bases and root rot symptoms. The losses 

caused by this pathogen, depended on species of cultivated plants and varied from 

10% to even 50%. 

Ours results indicated on considerable threat of P. cryptogea to plant crops in 

ornamental hardy nursery stocks as well as in greenhouses. The studies showed the 

lack of specialization specific for host plant and pathogen. P. cryptogea is not a new 

pathogen in Polish horticulture. The species was earlier isolated from rooted stems 

rot of cineraria, pachypodium and pelargonium (Orlikowski et al., 1984; Orlikowski, 

1996; 2003). In the last decade of XX century P. cryptogea was the reason of root 

and stem rot of Abies alba, Pinus mugho var. pumilo and P. nigra (Orlikowski et al., 

1995). In the beginning of XXI century the species was also detected from diseased 

Chamaecyparis lawsoniana (Szkuta, 2004). In the last two years P. cryptogea 

was found in perennial nurseries on Aquilegia, Sempervivum and Saxifraga species 

(Orlikowski, Ptaszek, 2007) and also was isolated from rotted stem base of Forsythia 

intermedia (Orlikowski, Ptaszek, 2008). In Germany, P. cryptogea was detected in 

water and sediments of hardy ornamental nursery reservoirs (Themann et al., 2002). 

The pathogen was also recovered from Polish water ponds and drainage canals situated 

in ornamental nurseries (Orlikowski unpbl.). It is possible that the species will spread 

on other plants till now not known as pathogen hosts. 

Conclusions. Phytophthora cryptogea was the most often isolated species from 

diseased A. discolor, A. x hybrida and S. arachnoideum as casual agent of stem/leaf 

bases and root rot. 

162

The species was isolated first time in Poland from diseased alstroemeria, forsythia 

and coniferous plants and some perennials including Aquilegia Sempervivum and 

Saxifraga. 

Results of laboratory trials showed significant differences in pathogenicity of 

P. cryptogea isolates toward alstroemeria, columbine and houseleek. 

Gauta 2009 06 30 


References 

1. Boersma J. G., Cooke D. E. L., Sivasithamparam K. 2000. A survey of wildflower 

forms in the south-west of Western Australia for Phytophthora spp. associated 

with root rots. Aust. J. Exp. Agric., 40: 1 011–1 019. 

2. Erwin D. C., Ribeiro O. K. 1996. Phytophthora DiseasesWorldwide. The APS, 

St. Paul. Minesota. 562. 

3. Orlikowski L. B. 1976/77. Przyczyny zamierania gerbery w niektórych gospodarstwach 

ogrodniczych w Polsce. Prace Inst. Sadownictwa Seria B, 2: 197–201. 

4. Orlikowski L. B. 1996. Phytophthora stem rot of Pelargonium. Phytopathol. Pol. 

12: 79–86. 

5. Orlikowski L. B. 1996. Phytophthora species in Polish ornamental nurseries. II. 

Chemical and biological control of P. cinnamomi on Chamaecyparis lawsoniana 

cv. Ellwoodii. Phytopathol. Pol., 11: 111–120. 

6. Orlikowski L. B. 2003. Phytophthora stem rot of Pachypodium lameri and its 

control. Phytopathol. Pol., 5: 17–21. 

7. Orlikowski L. B., Gabarkiewicz R., Skrzypczak C. 1995. Phytophthora species 

in Polish ornamental nurseries. I. Isolation and identyfication of Phytophthora 

species. Phytopathol. Pol., 9: 73–79. 

8. Orlikowski L. B., Ptaszek M. 2007. Phytophthora spp. in Polish ornamental nurseries. 

I. Perennial plants, new hosts of P. cryptogea. J. Plant Prot. Res., 47(4): 

401–408. 

9. Orlikowski L. B., Ptaszek M. 2008. Phytophthora cryptogea and P. citrophthora; 

new pathogens of Forsythia intermedia in Polish ornamental hardy nursery 

stocks. J. Plant Prot. Res., 48(4): 511–517. 

10. Orlikowski L. B., Skrzypczak C., Wojdyła A. 1984. Occurence, biology, pathogenicity 

and control of Phytophthora cryptogea on cineraria. Prace Instytutu 

Sadownictwa, Seria B, 9: 79–85. 

11. Orlikowski L.B., Szkuta G. 2002 a. Occurence of Phytophthora citrophthora on 

Syringa vulgaris in Poland. Acta Mycol., 40: 175–180. 

12. Orlikowski L. B., Szkuta G. 2002 b. Occurence of Phytophthora cinnamomi on 

ericaceous plants in container-grown ornamental nurseries in Poland. J. Plant 

Prot. Res., 42(2): 157–163. 

163

13. Orlikowski L. B., Szkuta. 2003. Phytophthora citricola on rhododendron spp. in 

Polish nurseries. J.Plant Prot. Res., 43 (1): 19-24. 

14. Orlikowski L. B., Szkuta G., Sroczyński M. 2004. First notice of Phytophthora 

tip blight of Calluna vulgaris. Phytopathol. Pol,. 31: 67–71. 

15. Szkuta G. 2004. Występowanie, izolacja, identyfikacja i szkodliwość gatunków 

z rodzaju Phytophthora w szkółkach ozdobnych roślin iglastych. Praca doktorska 

. Akademia Rolnicza w Krakowie, 191. 

16. Themann K., Werres S., Luttmann R., Diener H. A. 2002. Observations of 

Phytophthora spp. in water recirculation systems in commercial hardy ornamental 

nursery stock. Europ. J. Plant Pathol., 108: 337–343. 


Nauji augalai Phytophthora cryptogea vystymuisi Lenkijoje 

M. Ptaszek, L. B. Orlikowski, C. Skrzypczak 

Santrauka 

Tirtas devynių Phytophthora cryptogea izoliatų iš skirtingų augalų patogeniškumas. 

Laboratorijos tyrimuose visos kultūros (išskyrus S. arachnoideum) kolonizavo 3 Alstroemeria 

veislių lapalakščius ir stiebus, tačiau patogeniškiausi buvo izoliatai iš Anthurium andreanum, 

Aquilegia discolor ir Alstroemeria × hybrida. Tyrimuose su sinavadais pastebėta, jog greičiausiai 

nekrozė plinta ant lapų, inokuliuotų su P. cryptogea nuo A. discolor ir Gerbera jamesonii, 

o lėčiausiai, kai buvo naudojami izoliatai iš Sempervivum arachnoideum. S. arachnoideum 

inokuliacija aštuoniais P. cryptogea izoliatais sukėlė lapų nekrozę, bet patogeniškiausi buvo 

izoliatai iš A. discolor, Campanula persicifolia ir S. arachnoideum. 

Reikšminiai žodžiai: izoliacija, kolonizacija, paplitimas, patogeniškumas, Phytophthora 

cryptogea, užkrėsti augalai. 

164




Occurrence of RBDV in Latvia and virus elimination 

in vitro by chemotherapy 

Neda Pūpola 1 , Līga Lepse 2 , Anna Kāle 1 , 

Inga Moročko-Bičevska 1 

1 

Latvia State Institute of Fruit-Growing, Graudu str. 1, Dobele, LV-3701, Latvia, 

e-mail neda.pupola@lvai.lv 

2 

Pūre Horticulture Research Centre, Abavas str. 2, Pūre, LV-3124, Latvia 

To determine the incidence and distribution of Raspberry bushy dwarf idaeovirus 

(RBDV) in Latvia 27 commercial and varietal collection plantations of Rubus spp. were 

surveyed in the spring of 2007. In total 224 leaf samples from 59 genotypes were collected 

for analyses. A combination of meristem tip culture with different antiviral compounds was 

used to test virus elimination possibilities in vitro from naturally infected plants of cultivar 

‘Babje Leto 2’. Plant samples for RBDV infection and the efficiency of virus elimination were 

verified by double-antibody sandwich enzyme-linked immunosorbent assay (DAS ELISA) 

using polyclonal antibodies. The obtained results showed that RBDV was spread in 70 % of 

surveyed raspberry plantations. The incidence of RBDV in the tested plants was 35 % and 

varied greatly among the cultivars. Most of the commonly grown cultivars from Eastern Europe, 

such as ‘Kirzach’, ‘Balzam’ and ‘Sputnica’, were infected with RBDV. Virus was not detected 

in plant samples of cultivar ‘Tulameen’. RBDV elimination combining meristem culture with 

ribavirin for all treated plants was unsuccessful. Treatment with azacytidine and dicyanamide 

was effective only for meristem clones originated from one mother plant. It suggests that the 

particular plants were infected with a stable virus isolate, which cannot be eliminated with 

chemotherapy and in vitro propagation techniques. To develop effective RBDV elimination 

procedures more work is necessary to characterize the virus isolates infecting raspberry and 

to optimise in vitro techniques. The experiments are being continued. 

Key words: antiviral compounds, chemotherapy, DAS ELISA, meristem culture, 

Rubus spp. 

Introduction. Red raspberry (Rubus idaeus L.) is one of the most important 

small fruit crop in Latvia, which occupies about 10 % of commercial fruit plantation 

area. Raspberry viruses are wide spread and they cause severe losses of yield 

and quality everywhere raspberry is cultivated. Raspberry bushy dwarf idaeovirus 

(RBDV) infects wild and cultivated Rubus spp. plants throughout the world and is 

one of the most important pathogens of red raspberry (Natsuaki et al., 1991). Every 

year viral diseases cause significant raspberry yield loses due to premature defoliation, 

decreased vigour, leaf curling, necrosis, abortion of drupelets, death of lateral 

165

shoots and increased winter kill (Mavrič Pleško et al., 2009). In some red raspberry, 

such as R. idaeus var. idaeus L. and R. idaeus var. strigosus Maxim, RBDV induces 

yellows disease, crumbly fruit and decrease of vigour. RBDV isolates are categorised 

into two groups: Scottish strain (RBDV-S) and resistance breaking strain (RBDV- 

RB). Most isolates studied worldwide fall into S group strain and some raspberry cultivars 

are resistant to this strain. Whereas RBDV-RB strain can infect raspberry cultivars 

that are immune or highly resistant to S type strain (Jones et al., 2000). RBDV 

is efficiently transmitted via seed and pollen. Other ways of natural transmission are 

not known. Infected wild and cultivated raspberries act as virus natural reservoirs 

(Wang et al., 2008). The only way to decrease RBDV spread in raspberry plantations 

is to use healthy planting material and resistant cultivars. Thermotherapy, chemotherapy, 

and tissue culture techniques have been used either alone or in combination 

to eliminate viruses from plants (Spiegel et al., 1993). The meristem-tip culture has 

been used widely for the production of virus-free plant material in many species 

propagated mainly by vegetative means (Manganaris et al., 2003). The chemotherapy 

is one of the newest methods for elimination of plant viruses and is widely used in 

combination with micro-propagation (Cieslinska, 2003). Compounds such as ribavirin, 

5-azacytidine and other antiviral compounds have been successfully utilized for 

virus elimination in other economically important crops (Nascimento et al., 2003). 

The certification program for raspberry planting material has not been established 

in Latvia. Therefore, the risk that RBDV has spread uncontrolled in raspberry plantations 

with infected planting material and by natural transmission during the long period 

of time is very high. Previous research on raspberry viruses was carried out in 1970s 

and was based only on visual observations and biological indexing. Nowadays data are 

not available about the spread of viral diseases in raspberry plantations including such 

important pathogen as RBDV. The aim of this research was to determine the incidence 

of RBDV in raspberry plantations and to investigate RBDV elimination possibilities 

by combining micro-propagation and chemotherapy techniques. 

Object, methods and conditions. S u r v e y s a n d p l a n t s a m p l i n g. 

During the spring of 2007 twenty seven commercial and varietal collection plantations 

of red raspberry and blackberry were surveyed in all regions of Latvia. In total 

224 leaf samples were collected from 59 genotypes. From them 60 samples were 

collected in varietal collections. Leaflets were collected randomly from each cultivar 

in rows approximately ¼, ½ and ¾ of the way across the fields (Strik, Martin, 2003). 

From plants with visible symptoms additional samples were collected. The samples 

were transported to the laboratory in ice bag, proceeded immediately for analyses or 

stored at −80 °C. 

M i c r o p r o p a g a t i o n a n d c h e m o t h e r a p y. To test the virus elimination 

possibilities in vitro 23 plants of raspberry cultivar ‘Babje Leto 2’ were selected 

from field trial at Pūre Horticulture Research Centre. Plant initiation in vitro was carried 

out by using meristem tip explants from the apical and lateral buds of naturally 

infected plants with RBDV. Modified M&S basal salt medium (Murashige, Skoog, 

1962) containing ¼ of nitrates and double strength Fe salts, supplemented with 170 

mg L -1 KH 2 

PO 4 

and 0.4 mg L -1 thiamine, 2 mg L -1 BAP, 0.05 mg L -1 ISS and 0.1 mg L -1 

GA 3 

were used for initiation of microplants. The microplants were cultivated in 

vitro for five passages in above described medium containing BAP 1 mg L -1 . For 

166

chemotherapy three variants of media supplemented with antiviral chemicals were 

compared. Antiviral compounds used were ribavirin 30 mg L -1 (Sharma et al., 2007), 

azacytidine 25 mg L -1 (Nascimento et al., 2003) and dicyanamide 25 mg L -1 (Bittner 

et al., 1989). The previously used propagation medium without antiviral compounds 

was used as a control. Twenty microplants were used in each treatment. The effect of 

antiviral compounds on microplants was recorded as a number of survived plants after 

25 days. The data from the chemotherapy treatments were subjected to analysis of 

variance (ANOVA) and mean values were compared using less significant difference 

(LSD) at 95 % significance level. 

R B D V d e t e c t i o n. For the detection of RBDV in plant material commercially 

available double-antibody sandwich enzyme-linked immunosorbent assay (DAS 

ELISA) kit (Bioreba AG, Switzerland) was used in all investigation steps according 

to the manufacturer instructions with some modifications. The coating and conjugate 

conditions were changed from manufacturer standard procedure of 4 h incubation at 

30 °C to an overnight incubation in the refrigerator at 4–6 °C. The absorbance was 

read at 405/492 nm with dual filter microplate reader (Asys Expert 96, Hitech, Austria) 

after 30 min, 1 h and 2 h incubation. A “cut-off” value was calculated according to 

manufacturer technical information (Bioreba AG, Switzerland). 

Results. During the surveys in Rubus spp. plantations chlorotic spots on leaves, 

yellowing, mottling, undersized height and crumbly fruits were observed. The presence 

of RBDV by DAS ELISA was detected in 70 % of surveyed plantations. From 

three surveyed raspberry farms all the tested samples were infected with RBDV. The 

incidence of RBDV in plant samples was 35 %. 

Raspberry cultivars ‘Balzam’ and ‘Sputnitsa’ were the most infected with RBDV 

among the other tested cultivars, but all the samples from cv. ‘Tulameen’ were negative 

with DAS ELISA (Fig.). 

Fig. Occurrence (%) of RBDV in tested raspberry samples 

Pav. RBDV paplitimas tirtuose aviečių pavyzdžiuose, % 

167

For the tests for virus elimination possibilities in the field selected raspberry 

plants of cultivar ‘Babje Leto 2’ showed typical symptoms of RBDV infection, such 

as, undersized height, small, yellow foliages and crumbly fruits. According to DAS 

ELISA test results, it was proved that nine raspberry plants are infected with RBDV. 

Infected raspberry plants with typical RBDV symptoms and positive reaction in DAS 

ELISA test were propagated in vitro for five passages and obtained microplants were 

tested for RBDV infection after third and fifth passage. After third passage RBDV was 

detected in meristem clones originated from two mother plants, but after fifth passage 

RBDV was detected in meristem clones from four mother plants (Table 1). 

Table 1. RBDV in raspberry meristem clones after third and fifth passage in vitro 

1 lentelė. RBDV aviečių meristeminiuose klonuose po trečiojo ir penktojo persodinimo 

in vitro 

Mother plant No. 

Motininio augalo Nr. 

3 rd passage* 

Trečiasis persodinimas 

5 th passage* 

Penktasis persodinimas 

1.13 - - 

2.18 + + 

2.19 + + 

2.21 - - 

3.13 - + 

3.14 - - 

3.15 - + 

* + RBDV detected by DAS ELISA; - RBDV not detected by DAS ELISA test 

* + RBDV aptikta DAS ELISA testu; - RBDV neaptikta DAS ELISA testu 

To improve RBDV elimination from infected microplants, infected meristem 

clones from three mother plants were treated by chemotherapy in mediums amended 

with different antiviral compounds. After 25 days different reaction of plants to the 

amendment of antiviral compounds in the medium was observed. The significantly 

highest amount of necrotic plants was observed in the medium containing ribavirin. 

The percentage of survived plants in other two media containing antiviral chemicals 

was high and did not differ significantly from control medium (Table 2). 

Table 2. Amount of survived raspberry microplants (%) after 25 days of chemotherapy 

2 lentelė. Išlikusių aviečių mikro augalų kiekis (%) praėjus 25 dienoms po chemoterapijos 

Treatment 

Variantas 

Mother plant No. 

Motininio augalo Nr. 

2.18 2.19 3.15 

Control / Kontrolė 99 96 99 

Ribavirin 30 mg L -1 70 60 83 

Azacytidine 25 mg L -1 100 100 100 

Dicyanamide 25 mg L -1 98 100 100 

γ 0.05 

0.04 0.04 0.03 

168

After chemotherapy microplants were tested with DAS ELISA for RBDV infection. 

Meristem clones originated from mother plants 2.18 and 2.19 after chemotherapy 

showed positive results on RBDV (Table 3). 

Table 3. The efficiency of chemotherapy 

3 lentelė. Chemoterapijos efektyvumas 

Mother plants No. Control 

Motininio augalo Nr. Kontroė 

Ribavirin Azacytidine Dicyanamide 

2.18. + + + n/a 

2.19. + + + + 

3.15. + + - - 

The RBDV elimination with ribavirin, azacytidine or dicyanamide for those clones 

was unsuccessful. Meristem clones originated from mother plant 3.15 remained 

infected with RBDV after treatment with ribavirin, but treatment with azacytidine or 

dicyanamide was successful according to DAS ELISA test. 

Discussion. The research presented here demonstrated that RBDV is widespread 

in raspberry plantations in Latvia and that all commonly grown cultivars are infected. 

In the countries where certification programmes are not established, like in Chile, the 

incidence of RBDV is 35–68 % (Medina et al., 2006), what corresponds to the data 

obtained in this study. The percentage of infected plants in commercial plantations 

was lower than in varietal collections. Possibly it could be explained that nowadays 

farmers prefer to grow new varieties with Bu gene that are resistant to RBDV Scottish 

strain (RBDV-S). However, another RBDV-RB strain that widely occurs in central 

Europe, Russia and Siberia is able to infect cultivars that are resistant to S strain 

(Diekmann et al., 1994; Knight, Barbara, 1999). Although no information is available 

if Russian raspberry cultivars have gene Bu, our research showed that in Latvia 

common raspberry cultivars from Eastern Europe, such as ‘Kirzach’, ‘Balzam’ and 

‘Sputnica’, are highly infected with RBDV, probably with RBDV-RB strain. RBDV 

infection was not found in collected samples from raspberry cultivar ‘Tulameen’, 

which lack resistance gene Bu and in other European countries was found that ‘Tulameen’ 

is susceptible to both strains (Chard et al., 2001; Wood, Hall, 2001; Martin, 

1999). It indicates that this cultivar was probably introduced as virus-free stock in 

Latvia and had not yet been infected with RBDV in the fields. 

The RBDV elimination by combination of micro-propagation and chemotherapy 

did not give the expected results. As shown in other studies RBDV is difficult or 

impossible to eliminate from certain genotypes of raspberry by meristem tip culture 

(Wang, Valkonen, 2009). No one of the used antiviral compounds eliminated the virus 

in raspberry meristem clones from all the mother plants. Ribavirin has been demonstrated 

to give highest results in virus elimination in comparison with other antiviral 

chemicals (Nascimento et al., 2003). However, ribavirin was shown be effective in 

elimination fruit viruses from apple, raspberries and Prunus spp. (Cieslinska, 2007; 

Sharma et al., 2007). In our work better results were obtained in treatment by azacytidine 

and dicyanamide for meristem clones originated from mother plant 3.15. This 

could be explained by the combination of host and virus genotype as shown in other 

169

studies that the efficiency of virus elimination in host species differs depending on the 

virus and the host genotype (Wang et al., 2008). Possibly the meristem clones 3.15 

were infected with other virus strain than clones from mother plants 2.18 and 2.19. 

Obtained results suggest that plants were infected with stable virus strains, which 

cannot be readily eliminated with chemotherapy and micro-propagation techniques. To 

improve RBDV elimination effect it is necessary to combine tissue culture techniques 

with chemotherapy and thermotherapy. The results obtained in this study will be useful 

in the establishment of virus-free planting material propagation and certification 

program in the country. The work is being continued. 

Conclusions. 1. In Latvia commonly grown raspberry cultivars from Eastern Europe, 

such as ‘Kirzach’, ‘Balzam’ and ‘Sputnica’, are highly infected with RBDV. 

2. Raspberry cultivar ‘Tulameen’ possibly was introduced as virus-free stock in 

Latvia and had not yet been infected with RBDV in the fields. 

3. Antiviral compounds ribavirin, azacytidine and dicyanamide alone are not 

enough effective for RBDV elimination from raspberry in vitro. 

4. In this experiment treated meristem plants were infected with stable virus 

strain, which cannot be readily eliminated with micro-propagation techniques and 

chemotherapy. 

Gauta 2009 06 30 


References 

1. Bittner H., Schenk G., Schuster G., Kluge S. 1989. Elimination by chemotherapy 

of potato virus S from potato plants grown in vitro. Potato Research, 32 (2): 

175–179. 

2. Chard J., Irvine S., Roberts A. M. I., Nevison I. M., McGavin W. J., Jones A. T. 

2001. Incidence and distribution of Raspberry bushy dwarf virus in commercial 

red raspberry (Rubus idaeus) crops in Scotland. Plant Disease, 85 (9): 985–988. 

3. Cieslinska M. 2003. Elimination of Strawberry mottle virus (SMoV) from 

Fragaria virginiana UC-11 indicator plants by thermotherapy and chemotherapy. 

Phytopathologia polonica, 30: 51–59. 

4. Cieslinska M. 2007. Application of thermo- and chemotherapy in vitro for 

eliminating some viruses infecting Prunus sp. fruit trees. Journal of Fruit and 

Ornamental Plant Research, 15: 117–124. 

5. Diekmann M., Frison E. A., Putter T. (eds.) 1994. FAO/IPGRI Technical 

Guidelines for the Safe Movement of Small Fruit Germplasm. Food and 

Agriculture Organization of the United Nations, Rome/International Plant 

Genetic Resources Institute. 70–72. 

170

6. Jones A. T., McGavin W. J., Mayo M. A., Angel-Diaz J. E., Kärenlampi S. O., 

Kokko H. 2000. Comparisons of some properties of two laboratory variants of 

Raspberry bushy dwarf virus (RBDV) with those of three previously characterised 

RBDV isolates. European Journal of Plant Pathology, 106: 623–632. 

7. Knight V. H., Barbara D. J. 1999. A review of Raspberry bushy dwarf virus 

at Hri-East Malling and the situation on a sample of commercial holdings in 

England in 1995 and 1996. Acta Horticulturae, 505: 263–271. 

8. Manganaris G. A., Economou A. S., Boubourakas I. N., Katis N. I. 2003. 

Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture 

in nectarine. Plant Cell Reports, 22: 195–200. 

9. Martin R. R. 1999. Raspberry viruses in Oregon, Washington and British 

Columbia. Acta Horticulturae, 505: 259–262. 

10. Mavrič Pleško I., Viršček Marn M., Širca S., Urek G. 2009. Biological, serological 

and molecular characterization of Raspberry bushy dwarf virus from grapevine 

and its detection in the nematode Longidorus juvenilis. European Journal of 

Plant Pathology, 123: 261–268. 

11. Medina C., Matus J. T., Zuniga M., San-Martin C., Arce-Johnson P. 2006. 

Occurrence and distribution of viruses in commercial plantings of Rubus, Ribes 

and Vaccinium species in Chile. Ciencia e Investigacion Agraria, 33(1): 23–28. 

12. Murashige T., Skoog F. 1962. A revised medium for rapid growth and bioassays 

with tobacco tissue cultures. Physiologia Plantarum, 15(3): 473–497. 

13. Nascimento L. C., Pio-Ribeiro G., Willadino L., Andrade G. P. 2003. Stock indexing 

and Potato virus Y elimination from potato plants cultivated in vitro. 

Scientia Agricola, 60 (3): 525–530. 

14. Natsuaki T., Mayo M. A., Jolly C. A., Murant A. F. 1991. Nucleotide sequence 

of Raspberry bushy dwarf virus RNA-2: a bicistronic component of a bipartite 

genome. Journal of General Virology, 72: 2 183–2 189. 

15. Sharma S., Singh B., Rani G., Zaidi A. A., Hallan V., Nagpal A., Virk G. S. 

2007. Production of Indian citrus ringspot virus free plants of kinnow employing 

chemotherapy coupled with shoot tip grafting. Journal Central European 

Agriculture, 8(1): 1–8. 

16. Spiegel S., Frison E. A., Converse R. H. 1993. Recent developments in therapy 

and virus-detection procedures for international movement of clonal plant germ 

plasm. Plant Disease, 77(12): 1 176–1 180. 

17. Strik B., Martin R. R. 2003. Impact of Raspberry bushy dwarf virus on ‘Marion’ 

blackberry. Plant Disease, 87(3): 294–296. 

18. Wang Q., Cuellar W. J., Rajamäki M. L., Hirata Y., Valkonen J. P. T. 2008. 

Combined thermotherapy and cryotherapy for efficient virus eradication: relation 

of virus distribution, subcellular changes, cell survival and viral RNA degradation 

in shoot tips. Molecular Plant Pathology, 9(2): 237–250. 

19. Wang Q., Valkonen J. P. T. 2009. Cryotherapy of shoot tips: novel pathogen 

eradication method. Trends in Plant Science, 14(3): 119–122. 

20. Wood G. A., Hall H. K. 2001. Source of Raspberry bushy dwarf virus in Rubus 

in New Zeeland, and the infectibility of some newer cultivars to this virus. New 

Zealand Journal of Crop and Horticultural Science, 29: 177–186. 

171


Aviečių žemaūgiškumo viruso (RBDV) paplitimas Latvijoje ir 

viruso naikinimas in vitro chemoterapija 

N. Pūpola, L. Lepse, A. Kāle, I. Moročko-Bičevska 

Santrauka 

Siekiant nustatyti aviečių žemaūgiškumo viruso (RBDV) paplitimą ir pasiskirstymą 

Latvijoje, 2007 metų pavasarį buvo ištirtos 27 Rubus spp. komercinės ir veislių kolekcijų plantacijos. 

Analizei buvo surinkti 224 lapų pavyzdžiai priklausantys 59 genotipams. Siekiant ištirti 

viruso pašalinimo in vitro iš natūraliai užkrėstų veislės ‘Babje leto 2’ augalų galimybes, buvo panaudotas 

meristeminės kultūros ir skirtingų antivirusinių cheminių medžiagų derinys. Aiškinantis 

užkrėstumą šiuo virusu ir pastarojo pašalinimo veiksmingumą, augalų pavyzdžiai buvo patikrinti 

imunofermentiniu metodu (DAS-ELISA), panaudojant polikloninius antikūnius. Gauti rezultatai 

parodė, kad RBDV paplitęs 70 % tirtų aviečių plantacijų. RBDV paplitimas tirtuose augaluose 

siekė 35 % ir atskirose veislėse labai skyrėsi. Daugelis populiariausių Rytų Europos veislių, kaip 

‘Kirzach’, ‘Balzam’ ir ‘Sputnica’, buvo užkrėstos AŽV. Viruso nerasta veislės ‘Tulameen’ pavyzdžiuose. 

Mėginimas išnaikinti RBDV, derinant meristeminę kultūrą su ribavirinu terpėje, visuose 

apdorotuose augaluose buvo nesėkmingas. Apdorojimas azacitidinu ir dicianamidu paveikė tik 

tuos meristemų klonus, kurie buvo kilę iš motininio augalo. Tai rodo, kad kai kurie augalai buvo 

užkrėsti stabiliu viruso izoliatu, kurio chemoterapija ir in vitro dauginimo metodais išnaikinti 

neįmanoma. Norint sukurti veiksmingas RBDV naikinimo priemones, reikia tiksliai apibūdinti 

avietes užkrečiančias viruso atmainas ir optimizuoti in vitro metodus. Tyrimai bus tęsiami. 

Reikšminiai žodžiai: antivirusinės medžiagos, chemoterapija, DAS-ELISA, meristeminė 

kultūra, Rubus spp. 

172




Toxicity of biopesticides to green apple aphid in 

apple-tree 

Laimutis Raudonis, Alma Valiuškaitė, Laisvūnė Duchovskienė, 

Elena Survilienė 



During two-year trial there was examined the toxicity of biopesticides BioNature R2000 

(a. i. azadirachta indica 210 g/l, Pinus resinosa 180 g/l, Ricinus communis), Bioshower (a. i. 

100 % fatty acids) and Insecticidal Soap (a. i. 20 % fatty acids) to green apple aphid (Aphis pomi 

Deg.) in apple-tree at the Lithuanian Institute of Horticulture. BioNature R2000, Bioshower 

and Insecticidal Soap applied to A. pomi were moderately toxic after 3 days and very toxic after 

14 days. It is allowed to use biopesticides BioNature R2000, Bioshower and Insecticidal Soap 

in organic farming in many countries and according to the trial data they could be effectively 

applied for control of green apple aphid in apple growing. 

Key words: biopesticides, green apple aphid, toxicity. 

Introduction. European guidelines for fruit production require restrictions in 

pesticide use (Dickler, Schaefermeyer, 1991). On the other hand, increasing consumer 

concerns over the use of pesticides has lead growers to adopt more environmentally 

friendly Integrated Pest Management (IPM) or Organic Farming (OF) approaches. 

The Integrated Pest Management (IPM), which is based on selective toxicity 

of pesticides to the invertebrate pests and harmless to predatory mites and insects, 

became the most relevant strategy of plant protection (Gruys et al., 1980; Tuovinen, 

1992; Edland, 1994; Leake, 2000; Cuthbertson, Murchie, 2003). For the development 

of successful IPM and OF strategies, information is required on the direct effect of 

chemicals upon the invertebrate pests of a given ecosystem. The toxicity of pesticides 

on pests has been widely studied in many countries, using a range of different chemical 

pesticides at different rates on various developmental stages (Duso et al., 1992; 

Sterk et al., 1994; Kim, Yoo, 2002; Choi et al., 2003; Hardman et al., 2003; Cuthbertson, 

Murchie, 2003; Raudonis et al., 2004; Martínez-Villar et al., 2005). However, 

the existing active substances of pesticides are reviewed by European Commission 

and many old with detrimental effect active substances will be withdrawn from the 

market. 

173

The organic market has grown exponentially in Europe during the last ten years. 

However, the organic fruit industry has shown the lowest growth rates (1–5 % market 

share) in comparison with the other approaches. One major reason is the high production 

risk due to high pest pressure and lack of control means (Tamm, 2004). Only 

some biopesticides, like Insecticidal soap, Azadirachtin and etc. are allowed to use 

for pest control in organic farming (Compendium of UK Organic standards, 2006). 

Only few studies on toxicity of biopesticides to A. pomi were carried out (Castagnoli 

et al., 2002). 

The aim of this study was to investigate the toxicity of some biopesticides on A. 

pomi. 

Object, methods and conditions. Field trials were carried out on apple-trees 

under organic growing conditions at the Lithuanian Institute of Horticulture in 2005– 

2006. Biopesticides BioNature R2000 3.0 l ha l -1 (a. i. Azadirachtin 210 g l -1 , Pinus 

resinosa 180 g l -1 , Ricinus communis), Bioshower 5.0 l ha l -1 (a. i. 100 % fatty acids) 

and Insecticidal Soap 20.0 l ha -1 (a. i. 20 % fatty acids) were tested. Apple-trees by 

naturally occurring green apple aphid (Aphis pomi Deg.) were directly sprayed with 

spray mixture equivalent to 500 l/ha at 73 growth stage according to BBCH scale 

(Meier, 1997). 5 trees were sprayed and 3 trees assessed for each replicate. The treatments 

were repeated four times at random plot distribution. 

The number of living aphids were counted or estimated on 10 previously marked 

extension shoots per plot. The first assessment was made immediately before application 

at 73 growth stage, the second – 3 (73 growth stage) and the last 14 (75 growth 

stage) days after application. 

Mortality of aphids was calculated: x = 100 (1 - Ab/Ba) (x – mortality, %, A – 

number of aphids before spraying in untreated plot, B – before spraying in treated plot, 

a – after spraying in untreated plot, b – after spraying in treated plot). 

The mortality of aphids was explained according to the quantitative toxicity 

categories employed by the International Organization for Biological Control for 

assessment of pesticide toxicity to predatory and phytophagous mites in field trials: 

non-toxic (< 25 % mortality), slightly toxic (25–50 %), moderately toxic (51–75 %), 

very toxic (> 75 %) (Hassan et al., 1985). 

The number of aphids was compared among treatments in this study with a 

single factor analysis of variance (ANOVA). Specific differences were identified with 

Duncan’s multiple range test. 

Results. Tables 1 and 2 describes the effect and mortality of biopesticides to 

green apple aphid. In 2005 there was increase in the population of A. pomi in untreated 

plots. There were observed from 185 to 237 and from 82 to 96 A. pomi per 

10 shoots in 2005 and 2006, respectively. All the tested products (BioNature R2000, 

Bioshower and Insecticidal soap) significantly reduced mean numbers of A. pomi. 

The mortality of BioNature R2000 3.0 l ha -1 to A. pomi ranged from 67.0 to 78.5 %, 

Bioshower 5.0 l ha -1 – 68.4–83.1 % and Insecticidal soap 20.0 l ha -1 – 64.9–84.8 %. 

BioNature R2000, containing 210 g l -1 Azadirachtin was moderately and/or very toxic 

to A. pomi. Higher toxicity of BioNature R2000 to A. pomi was achieved 14 days 

after treatment than 3 days after treatment. Insecticidal soap and Bioshower were 

rated as moderately or very toxic to A. pomi. 

174

Table 1. Effects of biopesticides on survival of Green apple aphid (A. pomi) in 

apple-tree in 2005 

1 lentelė. Biopesticidų poveikis žaliųjų obelinių amarų (A. pomi) mirtingumui, 2005 m. 

Treatment 

Variantai 

Rate 

Norma 

(l ha -1 ) 

Mean number of aphids per 10 shoots 

Vidutinis amarų skaičius 10 ūglių 

before 

treatment 

A 

prieš 

apdorojimą 

3 days after 

treatment 

3 dienos po 

apdorojimo 

14 days after 

treatment 

14 dienų po 

apdorojimo 

Mortality 

Mirtingumas (%) 

3 days after 

treatment 

3 dienos po 

apdorojimo 

14 days after 

treatment 

14 dienų po 

apdorojimo 

Control 

- 185 a 153 b 111 b - - 

Nepurkšta 

BioNature R2000 3.0 220 ab 60 a 35 a 67.0 a 78.5 a 

Bioshower 5.0 237 b 62 a 24 a 68.4 a 83.1 a 

Insecticidal Soap 20.0 231 b 67 a 21 a 64.9 a 84.8 a 

Insekticidinis muilas 

Means followed by the same letter are not different significantly (P = 0.05) according to 


Tomis pačiomis raidėmis pažymėtos reikšmės pagal Dunkano kriterijų (P = 0,05) iš esmės nesiskiria 

Table 2. Effects of biopesticides on survival of Green apple aphid (A. pomi) in 

apple-tree in 2006 

2 lentelė. Biopesticidų poveikis žaliųjų obelinių amarų (A. pomi) mirtingumui, 2006 m. 

Treatment 

Variantai 

Rate 

Norma 

(l ha -1 ) 

Mean number of aphids per 10 shoots 

Vidutinis amarų skaičius 10 ūglių 

before 

treatment 

A 

prieš 

apdorojimą 

3 days after 

treatment 

3 dienos po 

apdorojimo 

14 days after 

treatment 

14 dienų po 

apdorojimo 

Mortality 

Mirtingumas (%) 

3 days after 

treatment 

3 dienos po 

apdorojimo 

14 days after 

treatment 

14 dienų po 

apdorojimo 

Control 

- 82 a 89 b 81 c - - 

Nepurkšta 

BioNature R2000 3.0 96 b 30 a 26 b 71.2 a 76.6 a 

Bioshower 5.0 95 ab 27 a 16 a 73.8 a 82.9 b 

Insecticidal Soap 20.0 91 ab 23 a 14 a 74.7 a 84.4 b 

Insekticidinis muilas 




Discussion. Little is known concerning the toxicity of biopesticides on pests in 

orchards. The toxicity of biopesticides on aphids has been studied in some experiments, 

using Insecticidal soap or products based on Azadirachtin (Schuster, Stansly, 

2000; Ahmad et al., 2003; Bostanian et al., 2005; Bostanian, Akalach, 2006; Karagounis 

et al., 2006; Kraiss, Cullen, 2008; Tremblay et al., 2008). However, studies 

showed controversial toxicity of biopesticides depending on pest or their predators 

life stage, species, abiotic factors or even different years (Imai et al., 1995; Schuster, 

175

Stansly, 2000; Ahmad et al., 2003; Karagounis et al., 2006; Kraiss, Cullen, 2008). 

BioNature R2000, containing 210 g l -1 Azadirachtin was moderately and/or very 

toxic to A. pomi. Little is known on toxicity of BioNature R2000 to A. pomi from 

other studies. Similar toxicity results on Dysaphis plantaginea Passerini have been 

obtained in Germany. Various preparations based on Azadirachta indica were highly 

efficient against aphids (Hummel, Kleeberg, 1997; Wyss, 1997). In our study higher 

toxicity of BioNature was achieved 14 days after treatment than 3 days after treatment. 

It can be explained that Azadirachtin does not kill directly and insects remain alive, 

but it stops insects to take up food. Only few days later insects become inactive, move 

uncoordinatedly and cannot molt successfully and in consequence it dies (Kleeberg et 

al., 1997). There are not published data on toxicity of Bioshower to A. pomi. However, 

both formulations of Bioshower and Insecticidal soap are based on fatty acids. Some 

authors have found that Insecticidal soap provided significant control of populations 

of D. plantaginea, A. pomi and Myzus persicae Sulzer or other aphid species (Lawson, 

Weires, 1991; Reuter et al., 1993; Imai et al., 1995; Karagounis et al., 2006; Kraiss, 

Cullen, 2008). Similar results were obtained in our test, whereas Insecticidal soap and 

Bioshower were rated as moderately or very toxic to A. pomi. 

Conclusions. In conclusion, the results of this study show that BioNature R2000, 

Bioshower and Insecticidal Soap are from moderately toxic to very toxic to A. pomi. 

The mortality of BioNature R2000 3.0 l ha -1 to A. pomi ranged from 67.0 to 78.5 %, 

Bioshower 5.0 l ha -1 – 68.4–83.1 % and Insecticidal soap 20.0 l ha -1 – 64.9–84.8 %. 

All the tested biopesticides are allowed to use in organic farming in many countries 

and according to the trial data could be effectively applied for control of A. pomi in 

apple growing. 

Gauta 2009 06 


References 

1. Ahmad M., Ossiewatsch H. R., Basedow T. 2003. Effects of neem-treated aphids 

as food/hosts on their predators and parasitoids. Journal of Applied Entomology, 

127(8): 458–464. 

2. Bostanian N. J., Akalach M. 2006. The effect of indoxacarb and five other insecticides 

on Phytoseiulus persimilis (Acari : Phytoseiidae), Amblyseius fallacis 

(Acari : Phytoseiidae) and nymphs of Orius insidiosus (Hemiptera : 

Anthocoridae). Pest Management Science, 62(4): 334–339. 

3. Bostanian N. J., Akalach M., Chiasson H. 2005. Effects of a Chenopodium-based 

botanical insecticide/acaricide on Orius insidiosus (Hemiptera : Anthocoridae) 

and Aphidius colemani (Hymenoptera : Braconidae). Pest Management Science, 

61(10): 979–984. 

176

4. Castagnoli M., Angeli G., Liguori M., Forti D., Simoni S. 2002. Side effects of 

botanical insecticides on predatory mite Amblyseius andersoni (Chant). Journal 

of Pest Science, 75(5): 122–127. 

5. Choi W., Lee S. G., Park H. M., Ahn Y. J. 2003. Toxicity of Plant Essential 

Oils to Tetranychus urticae (Acari: Tetranychidae) and Phytoseiulus persimilis 

(Acari: Phytoseiidae). Journal of Economic Entomology, 97(2): 553–558. 

6. Compendium of UK Organic standards. 2006. http://www.defra.gov.uk/farm/ 

organic/standards/ 

7. Cuthbertson A. G. S., Murchie A. K. 2003. The impact of fungicides to control 

apple scab (Venturia inaequalis) on the predatory mite Anystis baccarum and 

its prey Aculus schlechtendali (apple rust mite) in Northern Ireland Bramley 

orchards. Crop protection, 22: 1125–1130. 

8. Dickler E., Schafermeyer S. 1991. General principles, guidelines and standards 

for integrated production of pome fruits in Europe and procedures for endorsement 

of national or regional guidelines and standards. IOBC/WPRS Bulletin, 

14(3): 1–67. 

9. Duso C., Camporese P., Geest L. P. S. 1992. Toxicity of a number of pesticides 

to strains of Typhlodromus pyri and Amblyseius andersoni (Acari: Phytoseiidae). 

Entomophaga, 37: 363–372. 

10. Edland T. 1994. Side-effects of fungicide and insecticide sprays on phytoseiid 

mites in apple orchards. Norwegian Journal of Agricultural sciences, 17: 

195–204. 

11. Gruys P., Jong D. J., Mandersloot H. J. 1980. Implementation of integrated control 

in orchards. In: A. K. Minks, P. Gruys (eds.). Integrated control of insects 

pests in the Netherlands. Wageningen: Pudoc, 11–17. 

12. Hardman J. M., Franklin J. L., Moreau D. L., Bostanian N. J. 2003. An index for 

selective toxicity of miticides to phytophagous mites and their predators based 

on orchard trials. Pest Management Science, 59: 1324–1332. 

13. Hassan E., Oomen P. A., Overmeer W. P., Plevoets J. P., Reboulet J. N., 

Rieckmann W., Samsoe-Petersen L., Shires S. W., Staubli A., Stevensen J., 

Tuset J. J., Vanwetswinkel G., Zon A. Q. 1985. Standard methods to the test of 

side-effects of pesticides on natural enemies of insects and mites developed by 

the IOBC/WPRS Working Group “Pesticides and Beneficial Organisms”, OEPP/ 

EPPO Bulletin, 15: 214–255. 

14. Hummel E., Kleeberg H. 1997. New results on the practical application of 

NeemAzal-formulations. In: K. H. Hoffmann, W. Volkl (eds). Mitteilungen der 

deutschen gesellschaft fuer allgemeine und angewandte entomologie. Bremen, 

331–336. 

15. Imai T., Tsuchiya S., Fujimori T. 1995. Humidity effects on activity of insecticidal 

soap for the Green Peach Aphid, Myzus-Persicae (Sulzer) (Hemiptera, 

Aphididae). Applied Entomology And Zoology, 30(1): 185–188. 

16. Karagounis C., Kourdoumbalos A. K., Margaritopoulos J. T., Nanos G. D., 

Tsitsipis J. A. 2006. Organic farming-compatible insecticides against the aphid 

Myzus persicae (Sulzer) in peach orchards. Journal Of Applied Entomology, 

130(3): 150–154. 

177

17. Kim S. S., Yoo S. S. 2002. Comparative toxicity of some acaricides to the predatory 

mite, Phytoseiulus persimilis and the two spotted spider mite, Tetranychus 

urticae. BioControl, 47(5): 563–573. 

18. Kleeberg H., Hummel E., Tross R. 1997. Different strategies to control the rosy 

apple aphid Dysaphis plantaginea in organic apple growing. In K. H. Hoffmann, 

W. Volkl (eds.). Mitteilungen der deutschen gesellschaft fuer allgemeine und 

angewandte entomologie. Bremen, 223–226. 

19. Kraiss H., Cullen E. M. 2008. Efficacy and nontarget effects of reduced-risk 

insecticides on Aphis glycines (Hemiptera : Aphididae) and its biological control 

agent Harmonia axyridis (Coleoptera: Coccinellidae). Journal Of Economic 

Entomology, 101(2): 391–398. 

20. Lawson D. S., Weires R. W. 1991. Management of European red mite (Acari, 

Tetranychidae) and several aphid species on apple with petroleum oils and an 

insecticidal soap. Journal of Economic Entomology, 84(5): 1 550–1 557. 

21. Leake A. 2000. The development of integrated crop management in agricultural 

crops: comparisons with conventional methods. Pest Management Science, 

56(11): 950–953. 

22. Martínez-Villar E., Sáenz-De-Cabezón F. J., Moreno-Grijalba F., Marco 

V., Pérez-Moreno I. 2005. Effects of azadirachtin on the two-spotted spider 

mite, Tetranychus urticae (Acari: Tetranychidae). Experimental and Applied 

Acarology, 35(3): 215–222. 

23. Meier U. 1997. Growth stages of Mono- and Dicotyledonous plants. BBCH 

Monograph. Berlin: Blackwell Wissenschafts-Verlag. 

24. Raudonis L., Survilienė E., Valiuškaitė A. 2004. Toxicity of Pesticides to Predatory 

Mites and Insects in Apple-tree site under field conditions. Environmental 

Toxicology, 19 (4): 291–295. 

25. Reuter L. L., Toscano N. C., Perring T. M. 1993. Modification of feeding-behavior 

of Myzus-Persicae (Homoptera, Aphididae) by selected compounds. 

Environmental Entomology, 22(5): 915–919. 

26. Schuster D. J., Stansly P. A. 2000. Response of two lacewing species to biorational 

and broad-spectrum insecticides. Phytoparasitica, 28(4): 297–304. 

27. Sterk G., Creemers P., Merckx K. 1994. Testing the side effects of pesticides on 

the predatory mite Typhlodromus pyri (Acari, Phytoseiidae) in the field trials. 

IOBC/WPRS Bull, 17(10): 27–40. 

28. Tamm L., Haseli A., Fuchs J. G., Weibel F. P., Wyss E. 2004. Organic fruit production 

in humid climates of Europe: Bottlenecks and new approaches in disease 

and pest control. Acta Horticulturae, 638: 333–339. 

29. Tremblay B., Belanger A., Brosseau M., Boivin G. 2008. Toxicity and sublethal 

effects of an insecticidal soap on Aphidius colemani (Hymenoptera : Braconidae). 

Pest Management Science, 64(3): 249–254. 

30. Tuovinen T. 1992. Predatory mites in Finish apple orchards. Acta Phyt. Ent. 

Hungarica, 27(2): 609–613. 

31. Wyss E. 1997. Different strategies to control the rosy apple aphid Dysaphis 

plantaginea in organic apple growing. In: K. H. Hoffmann, W. Volkl (eds.). 

Mitteilungen der deutschen gesellschaft fuer allgemeine und angewandte entomologie. 

Bremen, 233–236. 

178


Biopesticidų toksiškumas žaliesiems obeliniams amarams 

L. Raudonis, A. Valiuškaitė, L. Duchovskienė, E. Survilienė 

Santrauka 

Lietuvos sodininkystės ir daržininkystės institute dvejus metus tirtas biopesticidų 

BioNature R2000 (a.i. azadirachta indica 210 g/l, Pinus resinosa 180 g/l, Ricinus communis), 

Bioshower (v. m. 100 % riebiosios rūgštys) ir insekticidinio muilo (v. m. 20 % riebiosios 

rūgštys) toksiškumas žaliesiems obeliniams amarams (Aphis pomi Deg.). BioNature 

R2000, Bioshower ir insekticidinis muilas praėjus 3 dienoms po purškimo buvo vidutiniškai 

toksiški žaliesiems obeliniams amarams, o praėjus 14 dienų – labai toksiški. Biopesticidai 

BioNature R2000, Bioshower ir insekticidinis muilas yra leidžiami naudoti ekologi - 

niuose ūkiuose kenkėjams naikinti daugelyje šalių. Mūsų tyrimai rodo, kad šie produktai 

yra efektyvūs nuo žaliųjų obelinių amarų ir gali būti naudojami ekologiškai auginant obelis. 

Reikšminiai žodžiai: biopesticidai, toksiškumas, žalieji obeliniai amarai. 

179




Integrated aproach of apple scab management using 

iMETOS warning system 

Laimutis Raudonis, Alma Valiuškaitė 



In 2007–2008 in field trials two different apple scab control strategies were compared: the 

current strategy – conventional disease management (CDM) and integrated disease management 

(IDM), according to scab infection periods. A new internet based scab warning system 

iMETOS was used for detection of infection periods and forecast of disease intensity at three 

levels: light, moderate and severe. According to CDM, apple-trees were sprayed 9 times per 

season. Scab warning system gave a possibility to optimize the use of fungicides against scab 

and to reduce to 7–8 instead 9 the total spray applications per season. CDM and IDM gave high 

scab control in apple-trees and there were not found any essential difference in scab incidence 

between two control strategies. An efficiency of IDM and CDM against disease incidence on 

leaves was 90.5–95.1 and 88.5–94.1 %, respectively. Efficiency against disease incidence on 

fruits raged from 95.1 to 95.6 and from 91.2 to 94.1 %, respectively. 

Key words: ascospores, conidia, conventional disease management, efficacy, infection, 

iMETOS warning system, integrated, . 

Introduction. Venturia inaequalis causal agent of apple scab is the most important 

apple disease and its control depends almost exclusively on frequent use of 

fungicides. 75 % of the pesticide use in apple production is related to control of 

fungal diseases, in which apple scab has a share of 70 % (Creemers, Laer, 2006). European 

agriculture faced the strict demands to decrease the use of pesticides in order 

to reduce any human or environmental hazards; therefore, the need for evaluation 

and reduction of the use of pesticides is expressed. To obtain the reduction of the use 

of fungicides without any significant damage to the crop, the control of apple scab 

should be based on registration of climatic data, scouting of biotic parameters, infection 

risks and simulation disease models. The bioecology of V. inaequalis has been 

widely studied in many countries. The development of infection risks of apple scab 

highly depends on apple cultivar susceptibility, inoculum of the pathogen in orchards, 

ascospore release and the influence of such parameters as air temperature, duration of 

leaf wetness, relative humidity or light (Gadoury et al., 1998; Stensvand et al., 1998; 

181

Li, Xu, 2002; Rossi et al., 2003; Holb et al., 2004; Holb et al., 2005; Carisse et al., 

2007). Based on air temperature duration of leaf wetness and other parameters apple 

scab models have been developed, evaluated and recently successfully used for the 

disease control in apple orchards. Met stations are equipped with sensors for registration 

and transmission of data about temperature, relative humidity, rainfall, leaf 

wetness and others for prediction of apple scab infection risks (Berrie, 1994; Butt, 

Xu, 1994; Bühler, Gesle, 1994; Rossi et al., 1999; Raudonis, 2002; Raudonis et al., 

2003; Holb et al., 2003; Xu, Robinson, 2005; Atlamaz et al., 2007; Rossi, Bugiani, 

2007). However, little is known about the effect of recently introduced iMETOS scab 

warning system. Therefore, the objective of this study was to evaluate the effects of 

iMETOS scab warning system for the use in integrated disease management. 

Object, methods and conditions. In 2007–2008 field trials were carried out 

in the orchards of Lithuanian Institute of Horticulture to compare the current apple 

scab control strategy – conventional disease management (CDM) and integrated disease 

management (IDM), according to scab infection periods. Internet based pest and 

disease of horticultural plants warning system iMETOS (G. Pessl, Austria) recorded 

rainfall, air temperature, relative humidity, leaf wetness and calculated infection periods 

according to Mills and Laplante at three levels. At the beginning of the season 

iMETOS calculates release of scab ascospores, later infection and conidia infection 

when degree-day accumulation (base = 0 °C) was 500 °C. Susceptible to scab apple 

variety ‘Lobo’ was sprayed when release of ascospores, ascospores or conidia light 

infection risk reached more than 70–80 %. CDM was based on prophylactic applications 

and apple-trees were sprayed at 10–14 days intervals. Plot size consisted at 

least of 5 trees, 4 replications at random plot distribution. Fastac 50 EC 0.4 l ha -1 at 

09 and 73 growth stages according to BBCH scale (Meier, 1997) was used for control 

of insect pests. 

Disease incidence was calculated: P = n/N · 100. (P – disease incidence, %, n – 

number of attacked leaves or fruits, N – total number of investigated leaves or fruits). 

Disease intensity was calculated: R = ∑ab × 100/NK; R – disease intensity; a – the 

number of leaves or fruits damaged the same level, b – score of the scale; ∑ – the 

sum numbers of damaged leaves or fruits of different scores; K – the highest score of 

the scale (5). Injures caused by fungal diseases were evaluated according to a 6 point 

scale: 0 – no disease symptoms detected on leaves or fruits, 5 – injured more 75 % 

of leaf or fruit area. 

Scab incidence and intensity on leaves and fruits was compared among treatments 

in this study with a single factor analysis of variance (ANOVA). Specific differences 

were identified with Duncan’s multiple range test. 

The trial was carried out according to trial plan as presented in Table 1. 

182

Table 1. Trial plan 

1 lentelė. Bandymo planas 

rate g, AI ha -1 of 

fungicides 

fungicidų v. m. 

norma g ha -1 

IDM 

infection risk 

infekcijos 

laipsnis 

Cyprodinil 100 % ascospore 

release 

750 g kg -1 , 150 g ha -1 askosporų 

plitimas 


750 g kg -1 , 150 g ha -1 infection 

užsikrėtimas 

askosporomis 

Trifloxistrobin 100 % conidia 



konidijomis 

Trifloxistrobin 

— “ — 

500 g kg -1 , 75 g ha -1 

Krezoksim-methyl 

— “ — 

500 g kg -1 , 100 g ha -1 

Dithianon 

— “ — 

700 g kg -1 , 70 g ha -1 

Dithianon 

— “ — 

700 g kg -1 , 70 g ha -1 

date 

data 

BBCH 

growth 

stage 

augimo 

tarpsnis 

CDM 

rate g, AI ha -1 of 

fungicides 

fungicidų v. m. 

norma g ha -1 

date 

data 

BBCH 

growth 

stage 

augimo 

tarpsnis 

IV.28 07 

1 2 3 4 5 6 7 

2007 

IV.28 07 Cyprodinil 

750 g kg -1 , 150 g ha -1 

Dithianon 

700 g kg -1 , 70 g ha -1 — “ — 

V.14 59 Cyprodinil 

750 g kg -1 , 150 g ha -1 V.10 57 

V.28 69 Trifloxistrobin 

500 g kg -1 , 75 g ha -1 V.22 67 

VI.14 73 Trifloxistrobin VI.04 71 

500 g kg -1 , 75 g ha -1 

VI.29 75 Krezoksim-methyl VI.18 73 

500 g kg -1 , 100 g ha -1 

VII.12 77 Dithianon VII.02 76 

700 g kg -1 , 70 g ha -1 

VII.29 81 Dithianon VII.12 77 

700 g kg -1 , 70 g ha -1 

VIII.12 85 Dithianon VII.26 81 

700 g kg -1 , 70 g ha -1 

– – – – Dithianon 

700 g kg -1 , 70 g ha -1 VIII.08 85 


release 

750 g kg -1 , 150 g ha -1 askosporų 

plitimas 




askosporomis 

Trifloxistrobin 100 % ascospore 

infection 

500 g kg -1 , 75 g ha -1 užsikrėtimas 

askosporomis 

IV.18 07 

2008 

IV.18 07 Cyprodinil 

750 g kg -1 , 150 g ha -1 

V.02 57 Cyprodinil 

750 g kg -1 , 150 g ha -1 V.2 57 

V.23 69 Trifloxistrobin 

500 g kg -1 , 75 g ha -1 V.16 67 

183



1 2 3 4 5 6 7 

Trifloxistrobin 100 % conidia VI.16 75 Trifloxistrobin V.29 71 



konidijomis 

500 g kg -1 , 75 g ha -1 

Krezoksim-methyl — “ — VI.30 77 Krezoksim-methyl VI.12 73 

500 g kg -1 , 100 g ha -1 500 g kg -1 , 100 g ha -1 

Dithianon 

— “ — VII.14 79 Dithianon VII.02 77 

700 g kg -1 , 70 g ha -1 700 g kg -1 , 70 g ha -1 

Dithianon 

— “ — VII.31 85 Dithianon VII.12 79 

700 g kg -1 , 70 g ha -1 700 g kg -1 , 70 g ha -1 

– – – – Dithianon VII.25 81 

700 g kg -1 , 70 g ha -1 

– – – – Dithianon 

700 g kg -1 , 70 g ha -1 VII.08 85 

Results. Apple scab ascospore release, ascospore and conidia infection periods, 

depending on air temperature, duration of leaf wetness and relative humidity 

in 2007–2008 are presented in Fig. 1 and 2. Apple trees were sprayed when release 

of ascospores, ascospores or conidia light infection risk reached more than 70–80 %, 

according to IDM using iMETOS warning system. In 2007 conditions for ascospore 

release reached 100 % on 26th of April (Fig. 1 a) and first fungicide application was 

made. Second application was made just after ascospore infection when it reached 

more than 85 % (Fig. 1 b). 

184

Fig. 1. Scab ascospore release (a), ascospore infections (b) and 

conidia infections (c) depending on climatic parameters in 2007 according to iMETOS. 

1 pav. Rauplių askosporų plitimas (a), užsikrėtimas askosporomis (b) bei 

konidijomis (c) priklausomai nuo oro sąlygų, pagal iMETOS prognozavimo sistemą 2007 m. 

Longer duration of leaf wetness resulted in more favourable conditions for ascospore 

release and infection in 2008. The duration of leaf wetness in April–May lasted 

9 055 and 14 015 min in 2007 and 2008, respectively (Fig. 1 a, b, 2 a, b). After flowering, 

when degree-day accumulation (base = 0 °C) was over 500 °C, scab conidia infections 

were led by next sprays. Warm weather and continuously lasting duration of leaf 

wetness often caused severe conidia infections during the summer of 2007 (Fig. 1 c). 

Only from the end of May till the middle of June and from the middle till the end of 

July there was no conidia infection observed. Less favourable weather conditions for 

conidia infections were in 2008. It can be explained by shorter duration of leaf wetness. 

The duration of leaf wetness in June–July lasted for 24 795 min in 2007 and near 

two times less (13 725 min) in 2008 (Fig. 1 c, 2 c). CDM was based on prophylactic 

185

applications and apple-trees were sprayed at 10–14 days intervals, depending only on 

growth stages. Higher number of sprays was made in CDM comparing with IDM strategy, 

when fungicides were applied only depending on scab infection. Nine fungicide 

treatments were applied in CDM strategy during both experimental years. Meanwhile, 

treatments were reduced to eight and seven in 2007 and 2008, respectively according 

IDM. Nevertheless, significant difference in scab incidence and intensity was not found 

on leaves and on fruits at harvest between both spray programmes (Table 2). In 2007 

there were 8.2 and 3.7 % scabbed leaves and fruits in IDM strategy and 9.5 and 6.7 % 

in CDM, respectively. In 2008 the damage on leaves and fruits was 2.0 and 3.75 % in 

IDM and 4.0 and 8.2 % in CDM, accordingly. Meanwhile, the leaves and fruits were 

damaged by 85 and 97.7 in 2007 and by 49.0–91.5 % in 2008 in unsprayed orchard. An 

efficiency of IDM strategy against apple scab incidence on leaves/fruits ranged from 

90.5/95.6 to 95.1/98.6 % and intensity from 90.4/95.9 to 95.9/96.2 %, respectively 

during 2007–2008. Similar efficiency results were obtained in CMD. 

186

Fig. 2. Scab ascospore release (a), ascospore infections (b) and 

conidia infections (c) depending on climatic parameters in 2008 according to iMETOS. 

2 pav. Rauplių askosporų plitimas (a), užsikrėtimas askosporomis (b) 

bei konidijomis (c) priklausomai nuo oro sąlygų, 

pagal iMETOS prognozavimo sistemą 2008 m. 

Table 2. Scab incidence and intensity under Integrated and Conventional Disease 

Management 

2 lentelė. Rauplių paplitimas ir intensyvumas taikant integruotą ir įprastinę apsaugą nuo ligų 

Treatments 

Variantai 

Apple scab 

Obelų rauplės (%) 

leaves 

lapai 

fruits 

vaisiai 

incidence 

paplitimas 

intensity 

intensyvumas 

incidence 

paplitimas 

intensity 

intensyvumas 

2007 

85.0 b 50.4 b 97.7 b 77.9 b 

Untreated 

Nepurkšta 

IDM 8.2 a 2.5 a 3.7 a 1.1 a 

CDM 9.5 a 3.0 a 6.7 a 2.2 a 

2008 

Untreated 

49.0 b 24.3 b 91.5 b 68.3 b 

Nepurkšta 

IDM 2.0 a 2.3 a 3.75 a 3.0 a 

CDM 4.0 a 2.8 a 8.2 a 6.0 a 




187

Fig. 3. Impact of Integrated and Conventional Disease Management on Apple scab incidence 

3 pav. Integruotos ir įprastinės apsaugos sistemų įtaka rauplių paplitimui 

Fig. 4. Impact of Integrated and Conventional Disease Management on Apple scab intensity 

4 pav. Integruotos ir įprastinės apsaugos sistemų įtaka rauplių intensyvumui 

188

Discussion. Air temperature, relative humidity and leaf wetness are the factors, 

which mostly influence the development of apple scab infections (Gadoury et al., 

1998; Hartman et al., 1999; Rossi, Bugiani, 2007; Laer, Creemers, 2004; Hamada, 

2005; Fröhling et al., 2005; Xu, Robinson, 2005). Refereeing these factors, scab 

warning systems for prediction of ascospores or conidia infection and scab control 

are developed (Berrie, 1994; Bühler, Gesle, 1994; Butt, Xu, 1994; Atlamaz et al., 

2007). In the orchards of the Lithuanian Institute of Horticulture IDM strategy was 

successful during experimental years, though climatically conditions for disease development 

were favourable. Scab incidence on fruits was increasing in unsprayed 

plots to 97.7 and 91.5 % in 2007 and 2008. With the improved apple scab warning 

system of the IDM, which is based on iMETOS scab warning system, it was possible 

to avoid one fungicide application in 2007 and two in 2008, comparing with CDM. 

Totally IDM strategy gave a possibility to optimize the use of fungicides against scab 

and to reduce on average to 7–8, instead 9 the total spray applications per season 

without any significant increase of damage to the fruits and their quality. High effect 

of the use of different apple scab warning systems have been reported in other studies 

(Berrie, 1994; Bühler, Gesle, 1994; Butt, Xu, 1994; Grauslund, Loshenkohl, 1994; 

Pessl, 1994; Raudonis, 2002; Holb et al., 2003; Raudonis et al., 2003; Atlamaz et al., 

2007; Rossi, Bugiani, 2007). 

Conclusions. A new internet based scab warning system iMETOS was used for 

detection of infection periods and forecast of disease intensity at three levels: light, 

moderate and severe. According to CDM apple-trees were sprayed 9 times per season. 

Scab warning system gave a possibility to optimize the use of fungicides against 

scab and to reduce to 7–8 instead 9 the total spray applications per season. CDM and 

IDM gave high scab control (88.5–95.1 %) in apple-trees and there were not found 

any essential difference in scab incidence between two control strategies. 

Gauta 2009 06 


References 

1. Atlamaz A., Zeki C., Uludag A. 2007. The importance of forecasting and warning 

systems in implementation of integrated pest management in apple orchards 

in Turkey. EPPO Bulletin, 37(2): 295–299. 

2. Berrie A. 1994. Practical experiences with the Ventem TM system for managed 

control of apple scab in the United Kingdom. Norwegian Journal of Agricultural 

Sciences. 17: 295–301. 

3. Bühler M., Gesle C. 1994. First experiences with an improved apple scab control 

strategy. Norwegian Journal of Agriculture Sciences. 17: 229–240. 

4. Butt D. J., Xu X. M. 1994. VentemTM – a comparison apple scab warning system 

for use on farms. Norwegian Journal of Agricultural Sciences. 17: 247–251. 

189

5. Carisse O., Rolland D., Savary S. . 2007. Heterogeneity of the aerial concentration 

and deposition of ascospores of Venturia inaequalis within a tree canopy 

during the rain. European Journal of Plant Pathology, 117(1): 13–24. 

6. Creemers P., van Laer S. 2006. Key strategies for reduction of the dependence 

on fungicides in integrated fruit production. Phytopathology, 39: 19–29. 

7. Gadoury D. M., Stensvand A., Seem R. C. 1998. Influence of light, relative humidity, 

and maturity of populations on discharge of ascospores of Venturia inaequalis. 

Phytopathology, 88(9): 902–909. 

8. Grauslund J., Loshenkohl. 1994. Control of apple scab according to warning 

equipment. Norwegian Journal of Agricultural Sciences. 17: 241–246. 

9. Fröhling P., Steiner U., Oerke E.C. 2005. Influence of temperature on pre- and 

post- infectional development of Venturia inaequalis isolates. Modern fungicides 

and antifungal compounds IV: 14th International Reinhardsbrunn Symposium, 

25–29 April, Friedrichroda, Thuringia, Germany, 193–199. 

10. Hamada, N. A. 2005. Influence of temperature and leaf wetness period (LWP) on 

the incidence and severity of gala leaf spot (Colletotrichum spp.). Agropecuária 

Catarinense, 18(2): 73–77. 

11. Hartman J. R., Parisi L., Bautrais P. 1999. Effect of leaf wetness duration, temperature, 

and conidial inoculum dose on apple scab infections. Plant Disease, 

83(6): 531–534. 

12. Holb I. J., Heijne B., Jeger M. J. 2003. Summer epidemics of apple scab: the relationship 

between measurements and their implications for the development of 

predictive models and threshold levels under different disease control regimes. 

Journal of Phytopathology, 151(6): 335–343. 

13. Holb I. J., Heijne B., Jeger M. J. 2004. Overwintering of conidia of Venturia 

inaequalis and the contribution to early epidemics of apple scab. Plant disease, 

88(7): 751–757. 

14. Holb I. J., Heijne B., Jeger M. J. 2005. The widespread occurrence of overwintered 

conidial inoculum of Venturia inaequalis on shoots and buds in organic 

and integrated apple orchards across the Netherlands. European Journal of Plant 

Pathology, 111(2): 157–168. 

15. Laer S. V. Creemers P. 2004. Preventive apple scab infection warnings: optimization 

of leaf wetness models and evaluation of regional weather forecast data. 

Proceedings of the IOBC/WPRS 6th International Conference on Integrated 

Fruit Production. 26–30 September, Baselga di Piné, Italy, 37–41. 

16. Li B., Xu X. 2002. Infection and development of apple scab (Venturia inaequalis) 

on old leaves. Journal of Phytopathology, 150(11–12): 687–691. 

17. Meier U. 1997. Growth stages of Mono- and Dicotyledonous plants. Berlin. 

18. Pessl G. 1994. Monitored and forecast weather for use on farm. Norwegian 

Journal of Agricultural Sciences. 17: 425–427. 

19. Raudonis L. 2002. Integrated control strategy of apple scab according to warning 

equipment. Plant Protection Science, 38(2): 700–703. 

190

20. Raudonis L., Valiuškaitė A., Rašinskienė A., Survilienė E. 2003. Pests and 

disease management model of apple-trees according to warning equipment. 

Sodininkystė ir daržininkystė, 22(3): 528–537. 

21. Rossi V. S., Bugiani G. R. 2007. A-scab (Apple-scab), a simulation model for 

estimating risk of Venturia inaequalis primary infections. EPPO Bulletin, 37 (2): 

300–308. 

22. Rossi V., Giosue S., Bugiani R. 2003. Influence of air temperature on the release 

of ascospores of Venturia inaequalis. Journal of Phytopathology, 151 (1): 

50–58. 

23. Rossi V., Ponti I., Marinelli M., Giosue S., Bugiani R. 1999. Field evaluation of 

some models estimating the seasonal pattern of airborne ascospores of Venturia 

inaequalis. Journal of Phytopathology, 147(10): 567–575. 

24. Stensvand A., Amundsen T., Semb L., Gadoury D. M., Seem R. C. 1998. 

Discharge and dissemination of ascospores by Venturia inaequalis during dew. 

Plant disease, 82(7): 761–764. 

25. Xu X. M., Robinson J. 2005. Modelling the effects of wetness duration and fruit 

maturity on infection of apple fruits of Cox’s orange Pippin and two clones of 

gala by Venturia inaequalis. Plant Pathology, 54(3): 347–356. 


Integruotas obelų rauplių valdymas naudojant iMETOS 

prognozavimo sistemą 

L. Raudonis, A. Valiuškaitė 

Santrauka 

2007–2008m. buvo palygintos dvi skirtingos obelų apsaugos nuo rauplių strategijos: 

įprastinė ir integruota obelų apsaugos sistema. Pastaroji pagrįsta nauja iMETOS prognozavimo 

sistema, kuri parodo obelų rauplių mažą, vidutinį ir didelį infekcijos rizikos lygį. 

Pagal tradicinę apsaugos nuo rauplių sistemą obelys buvo purkštos devynis kartus per sezoną. 

Rauplių prognozavimo sistema leido optimizuoti fungicidų panaudojimą nuo rauplių ir 

purškimų skaičių sumažinti iki septynių – aštuonių. Obelų rauplių paplitimas nesiskyrė panaudojus 

tiek tradicinę, tiek ir integruotą obelų apsaugos sistemą. Nustatytas integruotos ir tradicinės obelų 

apsaugos sistemų efektyvumas, rauplių plitimas ant lapų atitinkamai sumažėjo 90,5 – 95,1 ir 88,5 – 

94,1 %. Sistemų efektyvumas nuo rauplių ant vaisių atitinkamai buvo 95,1 – 95,6 ir 91,2 – 94,1. 

Reikšminiai žodžiai: askosporos, infekcija, iMETOS, integruota ir tradicinė apsaugos 

sistema, konidijos. 

191




First evidence of Itersonilia perplexans on dill 

(Anethum graveolens) in Bulgaria 

Rossitza Rodeva 1 , Jutta Gabler 2 , Zornica Stoyanova 1 

1 

Institute of Genetics, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria, 

e-mail rrodeva@yahoo.com 

2 

Institute for Epidemiology and Pathogen Diagnostics of the Julius Kühn-Institute 

(JKI) – Federal Research Center of Cultivated Plants, D-06484 Quedlinburg, 

Erwin-Baur-Str. 27, e-mail jutta.gabler@jki.bund.de 

A blight disease was detected on dill (cultivar ‘Dukat’) in private greenhouses in Bulgaria. 

The purpose of this investigation was to describe the symptoms of the disease, to identify 

the causal agent and to determine the pathogenicity and host range. Initial symptoms were 

small grey-green spots and wilting of leaf tips. Wilted leaves turned brown and collapsed as 

the disease developed. Necroses broadened so quickly that entire leaves dried within a short 

time. Foliage became a blighted making the leaves unsuitable for harvest. A fungus was 

consistently isolated from symptomatic leaves, petioles and stems of dill. The pathogen grew 

slowly on nutrient media and formed white to pale cream-colored colonies, velvety and flat 

with minimum aerial mycelium. The pathogenicity was confirmed on dill and other Apiaceae 

hosts. The fungus was identified as Itersonilia perplexans on the basis of colony morphology, 

hyphae with clamp connections and ballistospores. Disease caused by I. perplexans has not 

been found previously either on dill or any other host plants in Bulgaria up till now. 

Key words: Anethum graveolens, Apiaceae, Itersonilia perplexans. 

Introduction. The fungus Itersonilia perplexans Derx has been reported to be 

the causal agent of leaf blight of dill (Anethum graveolens L.) in several European 

countries, as Italy (Matta, Garibaldi, 1968), Germany (Geßner, 1988), Austria (Bedlan, 

1988), Swiss (Usoltseva, Dahl, 2006) as well as in USA (Koike, Tjosvold, 2001) 

and New Zealand (Anonymous, 2001; 2002; 2004; 2005; 2007). 

In the middle of June 2008 and in the end of May 2009, symptoms characteristics 

to I. perplexans infection were observed on dill grown in private greenhouses in 

Bulgaria. 

The purpose of this investigation was to describe the symptoms of the disease, to 

identify the causal agent and to determine the pathogenicity and host range. 

193

Object, methods and conditions. Infected dill plants (cultivar ‘Dukat’) were 

collected from private greenhouses in Sofia region. The causal agent was isolated 

from symptomatic leaves, leaf petioles and stems. Small pieces of diseased tissue 

were on the surface sterilized and plated on potato dextrose agar (PDA). The cultures 

were stored on cornmeal agar (CMA) slopes at 4 °C and subcultured every 

6 months. 

Three isolates obtained from leaf and stem tissue were selected to study cultural 

and morphological characteristics of the fungus. To determine radial growth rates of the 

isolates, 8 mm plugs were taken from the periphery of growing colonies and transferred 

to plates of PDA, CMA and 0.2 % malt extract agar (MEA). Inoculated plates were 

incubated at 22 °C and 12 h photoperiod. The diameter of each colony was measured 

4 and 16 days after inoculation. The rate of increase in colony radius was determined 

as the difference in colony size between 4 th and 16 th day after inoculation. There were 

four replicate plates. Morphological characters, including size of ballistospores were 

recorded for cultures grown for 2–3 weeks on three nutrient media. 

Ballistospores of each isolate were harvested by flooding the plates with sterile 

distilled water, scraping the plates with sterile paintbrush and filtering through double 

layers of sterile cheesecloth. Ballistospores were counted with a hematocytometer and 

adjusted to approximately 1 × 10 4 spores/ml. 

Potted seedlings of dill, coriander, celery, parsley, fennel, caraway and carrot were 

inoculated by spraying with ballistospore suspension. Inoculated plants were put in 

a humid chamber for 48 hours. Plants sprayed with sterile distilled water served as 

controls. Disease symptoms were evaluated 10 days after inoculation. Disease severity 

was rated on a 0–4 scale: 0 = no visible symptoms; 1 = small grey-green lesions; 

2 = brown lesions on leaves and petioles; 3 = leaves snapped off at the diseased lesions 

or 30–50 % of the leaf diseased; 4 = 50–100 % of the plant diseased. The data were 

processed by the McKinney’s formula (McKinney, 1923), which generates a numeric 

disease index (DI) of the severity of the attack: DI = (Σvn)/(NV) × 100, where v represents 

the numeric value of the class, n is the number of plants assigned to the class, 

N is the total number of the plants in the replication and V is the numeric value of the 

highest class. Reisolations were made from the diseased plant parts. 

Results. The symptoms on the naturally and artificially infected dill plants were 

characterized initially as small silvery-grey to grey-green spots distributed mainly on 

the leaf tips (Plate 1, a). Dark grey-green streaks were detected on leaf petioles and 

stems (Plate 1, b). The necroses broadened so quickly, that entire damaged leaves 

dried soon. Wilted leaves turned brown and collapsed (Plate 1, c). The foliage became 

blighted making the leaves unsuitable for harvest (Plate 1, d). The disease affected 

the umbel very seldom. 

A fungus was consistently isolated from symptomatic leaves, petioles and stems 

of dill. The colonies were slow growing. Average values for growth rates (mm/day) 

of three representative isolates used in the study on three nutrient media are presented 

in Fig. CMA nutrient medium assured the fastest growth of all isolates. Among them 

isolate 22 manifested the best growth on all nutrient media. 

194

Fig. Average growth rates of Itersonilia perplexans (isolates 21, 22 and 83) on 

three nutrient media (PDA, CMA and MEA) at 22 °C and 12 h photoperiod. 

Bars represent standard deviations (± SD). Least significant differences are 

0.05, 0.07, 0.09 for isolates and media and 0.09, 0.12, 0.16 for their interaction at 

P = 5 %, P = 1 %, P = 0.1 %, respectively. 

Pav. Vidutiniai Itersonilia perplexans (izoliatai 21, 22 ir 83) 

augimo greitis ant trijų mitybinių terpių (PDA, CMA ir MEA), 

esant 22 °C temperatūrai ir 12 h fotoperiodui. Stulpeliai atspindi standartinius 

nukrypimus (± SN). Mažiausi izoliatų ir terpės nukrypimai yra 0,05, 0,07, 0,09, 

o jų sąveikos – 0,09, 0,12, 0,16, kai atitinkamai P = 5 %, P = 1 %, P = 0.1 %. 

The studied isolates were all very similar, showing different colony morphology 

on three nutrient media (Plate II, a). The fungus grew slowly in 12 h photoperiod. In 

the dark the growth was still more stunted. The colony did not fill up the Petri dish in 

50 days (Plate II, b, c). 

On PDA the colonies were circular, more compact, pale cream-colored, velvety. 

The colonies on CMA and MEA were translucent, white and flat with minimal aerial 

mycelium (feathery margins on MEA). The mycelium was composed primarily of 

branched septate hyphae with prominent clamp connections at the septa (Plate III, 

a). Hyphae were hyaline, 2.7–4.5 μm wide and terminated in inflated sporogenous 

cells thin-walled, pyriform, ovoid to subglobose. These cells germinated to form 

hyphae or germ tubes, on which ballistospores formed. Ballistospores were lemonshaped, 

broadly-lunate, ovoid to pyriform with a basal flattering and a pointed tip, 

smooth-walled with granular content, (10) 14.12 ± 0.25 (17) × (7) 8.95 ± 0.16 (12) μm 

(Plate II, b) and germinated either with hyphae (Plate III, c) or secondary ballistospores. 

Chlamidospores were globose to subglobose, solitary or clustered, thick-walled, 

11–16 × 10–15 μm, produced mainly in old cultures (Plate III, d). 

Besides the usual method of tissue plantings, the fungus was isolated readily by 

spore shootings from surface disinfected diseased tissue suspended on the lids of Petri 

dishes over PDA. White spore deposits of typical ballistospores of I. perplexans were 

observed on agar plate under the suspended diseased plant material and numerous 

colonies developed. 

195

The fungus was identified as I. perplexans (Boekhout, 1991; Boekhout et al., 

1991). 

Inoculations proved the pathogenicity of the fungus. First symptoms were observed 

48 hours after inoculation. All tested isolates caused typical symptoms on dill – greygreen 

discoloration and wilting of leaf tips and streak lesions on leaf petioles and stems 

(DI = 79). The symptoms on fennel were similar but less severe than on dill (DI = 61). 

Small (1–2 mm in diameter) circular to lens-shaped, brown necrotic spots often surrounded 

by yellow haloes appeared on the leaf laminae of coriander (DI = 32). On 

carrot, caraway and parsley only single small spots were observed on the leaf periphery 

(DI = 25). The celery remained healthy. The fungus I. perplexans was consistently 

reisolated from the plant parts with symptoms. Water-treated control plants did not 

have any symptoms of disease and were negative to I. perplexans. 

Discussion. The causal agent was identified as I. perplexans based on colony 

morphology, hyphae with prominent clamp connections and ballistospores. The 

symptoms on artificially infected dill plants were similar to those on the naturally 

diseased ones observed in the greenhouses during June 2008 and May 2009. Some 

colonies of I. perplexans, free of contaminants, grew out from lesions but others 

were overrun by various fungi because of its slow growth. The fungus was isolated 

more readily by suspending diseased tissue on the lids of Petri dishes over PDA due 

to water-drop mechanism of ballistospore discharge. Lesions caused by I. perplexans 

have not been observed previously on dill plants in vegetable gardens and field 

experiments carried out at the Institute of Genetics, Sofia, during the last 10 years. 

Among the other representatives of Apiaceae family artificially inoculated fennel 

and coriander showed the most intensively developed disease symptoms. The fungus 

I. perplexans has been reported to be pathogenic on two main group plants belonging 

to Asteraceae (Sackston, 1958; McRitchie et al., 1973; McGovern, Seijo, 1999; Seijo 

et al., 2000; Horita, Yasuoka, 2002) and Apiaceae (Channon, 1963; Matta, Garibaldi, 

1968; Channon, 1969; Geßner, 1988; Bedlan, 1988, Koike, Tjosvold, 2001; Usoltseva, 

Dahl, 2006). The isolates showed specificity being pathogenic to the host group 

they were obtained from (Channon, 1963; Koike, Tjosvold, 2001; Horita, Yasuoka, 

2002). 

Conclusions. The disease caused by I. perplexans has not been recorded previously 

either on dill or any other host plants in Bulgaria up till now. Since I. perplexans 

grows best at high relative humidity the way to diminish the disease appearance and 

development is to keep the dill plants under as dry conditions as possible. The initial 

symptoms of I. perplexans resembled those caused by lack of nutrient substances or 

water. For this reason it is very important to clear up the cause as the first symptoms 

appear and to take control measures. The infected plants have to be destroyed and do 

not put in the compost. 

196

Acknowledgements. The investigations were carried out within the framework 

of the bilateral research project between Institute of Genetics, Bulgarian Academy 

of Sciences, Sofia, Bulgaria and Institute for Resistance Research and Pathogen Diagnostics, 

Federal Center for Breeding Research on Cultivated Plants, Aschersleben, 

Germany. Financial support by the Bulgarian National Science Fund (Project 

B-1301) is gratefully acknowledged. 

Gauta 2009 06 30 


References 

1. Anonymous. 2001. New organism records: 12/05/01–22/06/01. Biosecurity, 29: 

23–24. 

2. Anonymous. 2002. New organism records: 17/08/02–27/09/02. Biosecurity, 39: 

27–28. 

3. Anonymous. 2004. New organism records: 10/11/03 –12/12/03. Biosecurity, 49: 

18–19. 

4. Anonymous. 2005. Plant kingdom records: 18/12/2004–04/02/2005. Biosecurity, 

58: 21–23. 

5. Anonymous. 2007. Pest watch: 05/08/2007–14/09/2007. Biosecurity, 79: 27. 

6. Bedlan G. 1988. Erstmaliges Nachweis von Itersonilia perplexans Derv an Dill 

in Österreich. Pflanzenschutzberichte, 49 (1): 43–44. 

7. Boekhout T. 1991. Systematics of Itersonilia: a comparative phenetic study. 

Mycological Research, 95 (2): 135–146. 

8. Boekhout T., Poot G., Hackman P. 1991. Genomic characteristics of strains 

of Itersonilia: taxonomic sequences and life cycle. Canadian Journal of 

Microbiology, 37: 188–194. 

9. Channon A. G. 1963. Studies of parsnip canker. I. The causes of the disease. 

Annals of applied Biology, 51: 1–15. 

10. Channon A. G. 1969. Infection of the flowers and seed of parsnip by Itersonilia 

pastinacae. Annals of applied Biology, 64: 281–288. 

11. Geßner E. 1988. Blattspitzendürre an Dill: Erreger bekant – viele Frage offen. 

Phytomedizin. Mitteilungen der Deutschen Phytomedizinischen Gesellschaft, 

18(1): 10. 

12. Horita H., Yasuoka S. 2002. Black streak of edible burdock caused by Itersonilia 

perplexans in Japan. Journal of General Plant Pathology, 68: 277–283. 

13. Koike S. T., Tjosvold S. A. 2001. A blight disease of dill in California caused by 

Itersonilia perplexans. Plant Disease, 85(7): 802. 

14. Matta A., Garibaldi A. 1968. L’Itersonilia pastinacae Channon su Aneto. 

Phytopathologia Mediterranea, 7: 34–39. 

15. McGovern R. J., Seijo T. E. 1999. Petal blight of Callistephus chinensis caused 

by Itersonilia perplexans. Plant Disease, 83(4): 397. 

197

16. McKinney H. H. 1923. Influence of soil temperature and moisture on infection 

on wheat seedling by Helmintosporium sativum. Journal of Agricultural 

Research, 26(5): 195–217. 

17. McRitchie J. J., Kimbrough J. W., Engelhard A. W. 1973. Itersonilia petal blight 

of Chrisanthemum in Florida. Plant Disease Reporter, 57(2): 181–182. 

18. Sackston W. E. 1958. Itersonilia perplexans on sunflower in Uruguay. 


19. Seijo T. E., McGovern R. J., Marenco de Blandino A. 2000. Plant Disease, 

84(10): 1 153. 

20. Usoltseva M., Dahl Å. 2006. Förebygg svampangrepp i dill. Växtskydd, 9: 26. 


Pirmieji Itersonilia perplexans požymiai ant krapų (Anethum Graveolens) 

Bulgarijoje 

R. Rodeva, J. Gabler, Z. Stoyanova 

Santrauka 

Privačiuose Bulgarijos šiltnamiuose ant krapų (veislė ‘Dukat’) buvo aptiktos nekrozės. 

Šio tyrimo tikslas buvo apibūdinti ligos simptomus, išaiškinti sukėlėją ir nustatyti jo 

patogeniškumą bei augalus maitintojus. Pirmieji simptomai buvo mažos pilkšvai žalsvos 

dėmelės ir vystantys lapų galiukai. Ligai progresuojant, nuvytę lapai rudavo ir krito. Nekrozė 

plito taip greitai, kad per trumpą laiką nudžiūvo visi lapai ir nebetiko derliui. Grybas buvo 

izoliuotas nuo krapų lapų, žiedų ir stiebų su ligos požymiais. Ligos sukėlėjas iš lėto augo ant 

mitybinės terpės ir formavo nuo baltos iki pilkšvai kreminės spalvos kolonijas – aksomines 

ir plokščias, su minimaliu grybienos plotu. Patogeniškumas buvo patvirtintas krapuose ir 

kituose Apiaceae šeimos augaluose. Pagal kolonijos morfologiją, siūlinius grybus su gnybto 

tipo jungtimis ir balstosporas, nustatyta, jog tai Itersonilia perplexans. Patogeno I. perplexans 

sukeliama liga lig šiol nebuvo aptikta nei ant krapų, nei ant kokių nors kitų augalų Bulgarijoje. 

Reikšminiai žodžiai: Anethum graveolens, Apiaceae, Itersonilia perplexans. 

198




Investigation of Tobacco rattle virus infection in 

peonies (Paeonia L.) 

Marija Samuitienė 1 , Meletėlė Navalinskienė 1 , 

Stasė Dapkūnienė 2 

1 

Institute of Botany, Žaliųjų Ežerų 49, LT-08406 Vilnius, 

e-mail marija.samuitiene@botanika.lt 

2 

Plant Gene Bank, Vilnius University Botanical Garden, Kairėnų 43, LT-1023 

Vilnius, e-mail stase.dapkuniene@gf.vu.lt 

The aim of this study was to evaluate the occurrence of peony ring spot disease in the 

collections of foreign and Lithuanian origin peony cultivars and hybrids and to identify a 

causal agent of this viral disease. During surveys of ornamental plant collections grown in 

Lithuania, solitary instances of affected by peony ring spot disease plants have been detected 

in all collections surveyed. Diseased plants occurred in the similar frequency in cultivars of 

foreign and Lithuanian origin from species Paeonia lactiflora, P. lutea, P. officinalis, P. suffruticosa. 

Samples of diseased plants were collected for investigation of disease causal agent. 

Virus was isolated and identified using classical virological (test-plants, electron microscopy) 

and modern molecular biology (DAS-ELISA, RT-PCR) methods. Inoculated test-plants expressed 

reaction characteristic to Tobacco rattle virus (TRV); electron microscopy investigation 

revealed characteristic to this virus rod-shaped particles of two modal lengths. Data of 

DAS-ELISA and RT-PCR confirmed TRV infection in naturally infected peony plants and 

in inoculated test-plants. 

Key words: DAS-ELISA, RT-PCR, Tobacco rattle virus, test-plants. 

Introduction. Genus Paeonia L. has been attributed to Paeoniaceae family. 

There are 40 species in this genus of beautiful perennials and shrubs. The genus 

name goes back to classical Greek and arose from the mythological God Peon – the 

protector of human health and doctors. Peonies are all deciduous and have long-lived, 

rather woody rootstocks with swollen roots, and large compound leaves with leaflets 

usually toothed or lobed. Each stem in spring terminates in one to several large, roselike 

flowers. Their centers are the mass of short stamens that almost conceal the 2 to 

5 large ovaries, which develop into short pods containing large seeds. The flowers are 

mostly in shades of pink or red, but there are also white and yellow-flowered species. 

The great majority of peonies are herbaceous, dying back to the ground in autumn, 

but there is a small group of Chinese species, known as the tree peonies – shrubs 

(Cheifetz et al., 2004). 

199

Peonies have been grown for hundreds of years and remain very popular flower 

traditionally grown in every flower-garden of our country. Lithuanian breeders 

O. Skeivienė, E. Tarvidienė and J. Tarvidas carried out the breeding of Paeonia lactiflora 

Pall., S. Eicher-Lorka – of P. suffruticosa Andrews cultivars. Breeders have 

created many valuable peony cultivars and hybrids. Three cultivars of O. Skeivienė 

(P. lactiflora ‘Virgilijus’, ‘Garbė Motinai’, ‘Prof. K. Grybauskas’), four cultivars of 

S. Eicher-Lorka (P. suffruticosa ‘Elf’, ‘Žizel’, ‘Odeta’, ‘Otkrovenije’) and 25 hybrids of 

E. Tarvidienė and J. Tarvidas were selected and presented for conservation in Lithuanian 

Plant Genetic resources (Dapkūnienė, 2007; Dapkūnienė et al., 2008). 

Data on peony viral diseases are limited in literature. Tobacco rattle virus (TRV) 

infection has been reported in Japan (Chang et al., 1976), New Zealand (Jones, Young, 

1978) and China (Ningshen, Yunfeng, 1990). Strawberry latent ringspot and Raspberry 

ringspot viruses have been isolated from peony in Finland (Bremer, 1985), Alfalfa 

mosaic virus in Italy (Bellardi et al., 2003). First publication on peony ringspot disease 

in Lithuania was reported in 1974 (Макутенайте, 1974). Preliminary data on TRV as 

a causal agent of peony ringspot disease have been published in 1999 (Samuitienė, 

Navalinskienė, 1999). 

The aim of this study was to evaluate the occurrence of peony ring spot disease 

in the collections of foreign and Lithuanian origin peony cultivars and hybrids and to 

identify TRV as a causal agent of this disease. 

Object, methods and conditions. The plant material was collected in Botanical 

Gardens of Vilnius, Kaunas Vytautas Magnus Universities, and flower collection of 

The Lithuanian institute of Horticulture. 

According to Lithuanian State Program “Genefund” the virological evaluation 

of peony cultivars and hybrids created by Lithuanian breeders was carried out during 

period of 2004–2008. Lithuanian peony cultivars have been accumulated and grown 

in botanical gardens of Vilnius University (90 P. lactiflora cultivars and hybrids, 

originated by O. Skeivienė, E. Tarvidienė and J. Tarvidas); Kaunas Vytautas Magnus 

University (19 P. lactiflora cultivars and hybrids created by O. Skeivienė); flower collection 

of Lithuanian Institute of Horticulture (4 P. suffruticosa cultivars created by 

S. Eicher-Lorka). Peony plants grown in collections were surveyed once a year and 

tested for visual viral symptoms. 

Tobacco rattle virus has been identified by the methods of test-plant (Robinson, 

Harrison, 1989; Brunt et al., 1996), electron microscopy (EM) (Dijkstra, de Jager, 

1998), double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) 

(Clark, Adams, 1977) and reverse transcription polymerase chain reaction (RT-PCR) 

(Weidemann, 1995). 

The test-plants (Table) were inoculated in early stages of growth by mechanical 

sap transmission, applying carborundum as an abrasive. The inocula were prepared 

by homogenizing infected plant tissue in 0.1 M phosphate buffer pH 7.0, containing 

1 % nicotine acid as virus-stabilizing additive. 

Virus particles were examined in leaf dip preparations negatively stained with 

3 % uranyl acetate electron microscopically using a JEM-100S electron microscope, 

at the instrumental magnification of 25 000. 

200

DAS-ELISA was carried out using commercial kit (DSMZ Plant Virus Collection, 

Germany), according to standard procedure. TRV IgG and their conjugate with alkaline 

phosphatase were used at a dilution of 1/1 000. 50 mg of sample was extracted 

in 1 ml of sample buffer. 0.1 % p-nitrophenylphosphate was used as substrate. The 

reactions were measured after 90 min incubation with substrate photometrically at 

405 nm (Labsystems Multiskan RC). 

Reverse transcription polymerase chain reaction (RT–PCR) for TRV detection 

was accomplished using the primers designed from sequence of the 16 k protein gene 

of RNA–1 of TRV strain ‘SYM’. Oligonucleotide primer A was synthesized complementary 

to the nucleotides 6461 to 6478 5’–GGT GTC CCA AAT TCT CTG–3’ and 

oligonucleotide primer B in sense to orientation to nucleotides 6115 to 6133: 5’–CGT 

TGT GTA CTC AAG GGT TG–3’. Primers A and B define a target sequence of 364 bp 

(Hamilton et al., 1987; Weidemann, 1995). 

Total RNA was extracted from symptomatic test plant material stored frozen at 

-20 °C using QuickPrep TM Total RNA Extraction Kit (Amersham Biosciences UK). 

Extraction procedure was carried out according to the manufacturer’s instructions. 

All PCR procedures were carried out in Eppendorf Master Cycler Personal. For 

RNA denaturation mixture of 9 µl RNA (for each sample) and 1 µl 20 pM of downstream 

primer B was incubated 5 min at 70 °C and 5 min at 4 °C. 

Reverse transcription (RT, cDNA synthesis) reaction mixture (for one sample): 

4 µl 5 × PCR buffer; 1 µl RNasin; 2 µl 10 mM dNTPs; 1 µl 200 U/µl MulRev 

Transcriptase and 11 µl of denatured RNA mix. Reaction was performed incubating 

at 42 °C for 60 min, at 70 °C for 10 min and at 4 °C for 5 min. 

PCR reaction mixture contained (for one sample): 7 µl cDNA; 34.75 µl PCR water, 

4 µl 2 mM dNTPmix, 1 µl 20 pM primer A, 1 µl 20 pM primer B, 5 µl 10 × PCR buffer 

without Mg 2+ , 3 µl MgCl 2 

, 0.25 µl 5U/µl Taq DNA polymerase. Reaction mixtures 

were incubated at 94 °C for 5 min (for first step), 25 cycles of 94 °C for 45 sec, 49 °C 

for 46 sec, 72 °C for 1 min, and at 72 °C for 5 min (final step). 

Resulting PCR products were analyzed by electrophoresis through 5 % polyacrylamide 

gel, stained with ethidium bromide, and DNA bands visualized using a 

UV transilluminator. DNA fragment size standard was PhiX174 RFI DNA HaeIII 

digest, fragment sizes (from top to bottom): 1 353, 1 078, 873, 603, 310, 281, 271, 

234, 194, 118, 72 bp. 

Results. During regular surveys to monitor phytosanitary status of ornamental 

plants grown at Botanical gardens solitary instances of diseased plants from species 

Paonia lactiflora, P. lutea, P. officinalis, P. suffruticosa were observed in all collections 

surveyed. Peony plants showed symptoms characteristic of peony ring spot disease. 

At early stage of disease leaf tissue develops light green irregular-shaped spot, 

later becoming yellowish. Around them and also around green patches yellow rings 

appear. The pattern looks like concentric irregular rings and semi-rings. The pattern 

covers all leaf lamina. Symptoms make progress later in season, becoming particularly 

visible in autumn. The rings turned into large sinuous yellow bands. Considerable 

variation in symptom expression can be noticed depending on cultivar, growing 

conditions and season (Fig. 1). Some peony cultivars show chlorotic spots with green 

blotches in the middle, surrounded by yellow bands or rings with light-green dots. 

201

Fig. 1. Symptoms of ringspot disease on peony leaves 

1 pav. Žiediškosios dėmėtligės simptomai and bijūnų lapų 

Virus was isolated from collected samples of diseased peonies by the method of 

test-plants. Using mechanical inoculation the group of 17 test-plants representing five 

families was inoculated with virus isolates separated from naturally infected peonies. 

Test-plants and results of their reaction to inoculation are presented in Table. 

Table 1. Test-plant reactions to inoculation of virus isolated from peony 

1 lentelė. Augalų indikatorių reakcija užkrėtus iš bijūnų išskirtu virusu 

Test-plant 

Augalas indikatorius 

Reaction 

Reakcija 

1 2 

Aizoceaeae F. Rudolphi 

Tetragonia expansa Murr. 

L: NLL 

Amaranthaceae 

Amaranthus paniculatus L. 

L: BrLL 

Gomphrena globosa L. 

L: NSp 

Asteraceae Dumort 

Galinsoga parviflora Cav. 

L: NStr, RiSp 

Chenopodiaceae Vent. 

Atriplex hortensis L. 

L: LL 

Celosia argentea 

L: BrLL; S: LeDis,VB 

Chenopodium amaranticolor Coste et Reyn L: NLL 

C. ambrosioides L. L: NLL 

C. foetidum Schrad. L: LL 

C. quinoa Willd. L: Cl and NLL 

C. urbicum L. L: LL 

Cucurbitaceae Juss. 

Cucumis sativus L. 

L: Cl or NLL 

202

Table continued 

Lentelės tęsinys 

1 2 

Solanaceae Juss. 

Nicandra physalodes (L.) Gaertn. 

L: NLL; S: ClSp, RiSp 

Nicotiana debneyi Domin. 

L: L: NLL; S: LeDis, M 

N. glutinosa L. L:GNRi; S: NSp, NRi, Stu 

N. occidentalis Wheeler L: NRiSp; S: M, LeNr 

N. tabacum ‘Xanthi’ L. L: NSp; S: EtNRiPat, N, Dis, VC 

Abbreviations: L – local reaction; S – systemic reaction; LL – local lesions; Le – leaf; Cl – 

chlorotic, chlorosis; N – necrotic, necrosis; Stu – stunting; Sp – spots; M – mosaic; Nr – narrowing; 

Ri –rings; Dis – distortion; Str – streaks; C – clearing; Et – etching; Pat – pattern; 

Br – brown; G – grey; VB – vein banding; VC – vein clearing. 

Sutrumpinimai: L – vietinė reakcija; S – sisteminė reakcija; LL – vietiniai pažeidimai; Le – lapas; Cl – 

chlorozė, chlorotinis; N – nekrozė, nekrotinis; Stu – žemaūgė; Sp – dėmės; M – mozaika; Nr – susiaurėjimas; 

Ri – žiedai; Dis – deformacija; Str – dryžiai; C – pašviesėjimas; Et – graviruotė; Pat – piešinys; 

Br – rudas; G – pilkas; VB – apvadas apie gyslas; VC – gyslų pašviesėjimas. 

The resulting reaction of test-plants was typical for Tobacco rattle virus (TRV), 

described in literature (Robinson, Harrison, 1986; Brunt et al., 1996). Virus induced 

chlorotic and necrotic local lesions in all inoculated test-plants (Fig. 2). 

Fig. 2. Necrotic local lesions induced by TRV isolated 

from peony on leaves of inoculated test-plants: 

a – Nicandra physalodes; b – Tetragonia expansa 

2 pav. Vietiniai nekrotiniai pažeidimai sukelti iš bijūnų išskirtu 

TRV ant inokuliuotų augalų indikatorių lapų: 

a – Nicandra physalodes; b – Tetragonia expansa 

Celosia argentea and test-plants representatives of Solanaceae developed local 

and systemic reactions (Fig. 3). 

203

Fig. 3. Systemic reaction on test plants induced by TRV isolated from peony: 

a – vein banding and leaf distortion symptoms on Celosia argentea; 

b – necrotic ring etching on Nicotiana tabacum ‘Xanthi’ leaf 

3 pav. Sisteminė reakcija ant augalų indikatorių užkrėstų iš bijūnų išskirtu TRV: 

a – gyslų apvadų ir lapų deformacijos požymiai ant Celosia argentea; 

b – nekrotinė žiediškoji graviruotė ant Nicotiana tabacum ‘Xanthi’ lapo 

EM revealed rod-shaped virus particles of two modal lengths of 55–110 nm 

(short particles) and 200 nm (long particles) (Fig. 4). Such morphology of particles is 

characteristic of TRV (Robinson, Harrison, 1986; Brunt et al., 1996). 

Fig. 4. Electronomikrograph of TRV particles. Bar represents 100 nm. 

4 pav. TRV dalelių elektronomikrografija. Brūkšnys – 100 nm. 

Symptomatic host plants and inoculated test-plants were tested in DAS–ELISA. 

Reaction was considered positive when absorbance at 405 nm was higher than twice the 

mean of healthy (negative) controls. All tested plants gave clearly expressed positive 

reaction confirming TRV infection (data not shown). 

TRV identification by test-plant reactions, EM and serological test were verified 

in RT-PCR using as samples test-plants inoculated with virus isolated from peony 

(isolate No 0429). Total RNA was extracted from frozen leaf tissue of infected testplants. 

Leaf tissue from healthy Nicotiana tabacum plant was used as negative control 

(K-). Previously identified TRV isolate from gladiolus was used as positive control 

204

(K+). Specific for TRV PCR products were obtained with locally infected Tetragonia 

expansa, Chenopodium quinoa, systemically infected Celosia argentea, Nicotiana 

tabacum and positive control, but not with negative control. Specific bands in polyacrylamide 

gel of analyzed products after electrophoresis at a position corresponding 

to the expected size of amplification product of 364 bp were obtained, confirming 

TRV identity (Fig. 5). 

Fig. 5. RT-PCR products of amplified DNA fragments from TRV isolated from peony 

(EF 5 % PAGE). Lanes: 1, 8 – DNA size standard PhiX174 DNA HaeIII digest, fragment 

sizes (from top to bottom): 1 353, 1 078, 873, 603, 310, 281, 271, 234, 194, 118, 72 bp., 

2 – Tetragonia expansa; 3 – Chenopodium quinoa (local reaction); 4 – Celosia argentea; 

5 – Nicotiana tabacum (systemic reaction); 6 – (K-); 7 – (K+). Size of product – 364 bp. 

5 pav. Iš bijūnų išskirto TRV DNR fragmento pagausinto AT-PGR reakcijoje produktai 

(EF 5 % poliakrilamido gelyje). Takeliai: 1, 8 – DNR fragmentų dydžio standartas 

PhiX174 DNA HaeIII digest, fragmentų dydžiai (nuo viršaus į apačią): 

1 353, 1 078, 873, 603, 310, 281, 271, 234, 194, 118, 72 bp., 

2 – Tetragonia expansa 3 – Chenopodium quinoa (vietinė reakcija); 4 – Celosia argentea; 

5 – Nicotiana tabacum (sisteminė reakcija); 6 – K– ; 7 – K+. Produkto dydis – 364 bp. 

During surveying of Lithuanian peony cultivars in collections it was revealed that 

great majority of plants were healthy. One diseased plant in the cultivar ‘Garbė Motinai’ 

and two plants in hybrids were found showing symptoms of ring spot disease. These 

plants were removed in order to avoid spreading of infection. So, virological state of 

Lithuanian peony cultivars and hybrids growing in collections is good. 

Discussion. TRV was isolated from peony plants showing symptoms characteristic 

to peony ring spot disease. Virus was reliably identified using classical virological 

methods of test-plants, electron microscopy, and modern methods of molecular 

biology, DAS-ELISA, RT-PCR. TRV as the most prevalent agent of virus disease 

in peonies causing ring spot disease was described in literature (Chang et al., 1976; 

Jones, Young, 1978; Ningshen, Yunfeng, 1990; Brunt et al., 1996). TRV is the type 

member of Tobravirus genus, and it is the only virus of the genus to infect ornamental 

plants (Brunt, 1995). TRV has a very wide natural host range, including ornamentals; 

205

its infection was reported in ornamental species: Alstroemeria L., Lilium L., Narcissus 

L., Aster L., Crocus L., Freesia Eckl. Ex Klatt, Gladiolus L., Hyacinthus L., 

Iris L., Nerine Herb., Phlox L., Tulipa L. (Loebenstein et al., 1995). This virus is 

widespread on ornamentals in our country; it was isolated and identified in 15 species 

belonging to 10 botanical families (Samuitienė, Navalinskienė, 2000). More than 

400 species in more than 50 dicotyledonous and monocotyledonous families can be 

infected experimentally; in many instances the infection does not become systemic. 

The virus has rod-shaped particles c. 22 nm in diameter and of two predominant 

lengths 180–215 nm (long) and 46–115 nm (short). It is transmitted by Trichodorus 

and Paratrichodorus nematodes, species of which are strain specific vectors and can 

retains virus for many months if non-feeding. The virus is also seed-borne in some 

host species (Robinson, Harrison, 1989; Brunt, 1995; Brunt et al., 1996). 

Methods of controlling peony viruses consist of growing and multiplying selected 

healthy stocks, visual inspection of plants for symptoms and elimination of affected 

plants. The losses by nematode transmitted viruses can be reduced by means to control 

the nematodes. The application of meristem tip culture method can be applied for 

propagation virus-free plant material especially for the most valuable peony cultivars 

(Spiegel, Loebenstein, 1995). 

Conclusions. 1. Peony plants expressing symptoms characteristic to peony ring 

spot disease have been found as solitary instances in all collections surveyed in cultivars 

of foreign and Lithuanian origin from species Paeonia lactiflora Pall., P. lutea 

Delav. ex Franch., P. officinalis L., P. suffruticosa Andrews. 

2. The causal agent of peony ring spot disease was isolated and identified by the 

methods of test-plants, electron microscopy, DAS-ELISA and RT-PCR as Tobacco 

rattle virus (TRV). 

Acknowledgements. This work was supported in part by the grant No. 18 from 

Lithuanian State Program “Genefund” and by Lithuanian State Science and Studies 

Foundation (Project No. V–25/2009). 

Gauta 2009 06 30 


References 

1. Bellardi M. G., Rubies-Antonell C., Bianchi A. 2003. First report of a disease of 

peony caused by Alfalfa mosaic virus. Plant Disease, 87(1): 99. 

2. Bremer K. 1985. Strawberry latent ringspot virus in ornamental plants in Finland. 

Annales Agriculturae Fenniae, 24(2): 101–102. 

3. Brunt A. A. 1995. Major genera of plant viruses. In: Loebenstein G., Lawson R. H., 

Brunt A. A. (eds.). Virus and Virus-like Diseases of Bulb and Flower Crops. 

Wiley, Jerusalem: 29–66. 

206

4. Brunt A. A., Crabtree K., Dalwitz M. J., Gibbs A. J., Watson L. (eds.) 1996. 

Viruses of Plants. Descriptions and Lists from VIDE Database. UK Univ. Press, 

Cambridge. 1 484. 

5. Chang M. U., Doi Y., Yora K. 1976. A rod-shaped virus found in the peony ringspot. 

Annals of the Phytopathological Society of Japan, 42(3): 325–328. 

6. Cheifetz A., Double C., Barnard L., Imwold D. (eds.). 2006. Annuals and perennials. 

Tandem Verlag, Sydney. 

7. Clark M. F., Adams A. N. 1977. Characteristics of the microplate method of 

enzyme-linked immunosorbent assay for detection of plant viruses. Journal of 

General Virology, 34: 475–483. 

8. Dapkūnienė S. 2007. Lietuvos augalų nacionaliniai genetiniai ištekliai. 

Lietuviškos bijūnų veislės. Spaudvita, Kėdainiai. 

9. Dapkūnienė S., Motiejūnaitė O., Varkulevičienė J., Pakulienė J., Guseva, V., 

Mažeikienė I. 2008. Lietuviški bijūnai miestų gėlynams. In: Miestų želdynų 

formavimas’ 2008: gėlės ir gėlynai / Formation of Urban Green Areas’ 2008. 

Flowers and Paterres. Mokslinių straipsnių rinkinys, Klaipėdos verslo ir 

technologijų kolegija, Klaipėda. 

10. Dijkstra J., de Jager C. P. 1998. Practical Plant Virology. Protocols and Exercises. 

Springer-Verlag, Berlin. 

11. Hamilton W. D., Boccara M., Robinson D. J., Baulcombe D. C. 1987. The complete 

nucleotide sequence of tobacco rattle virus RNA-1. Journal of General 

Virology, 68: 2 563–2 575. 

12. Jones A. T., Young B. R. 1978. Some properties of two isolates of tobacco rattle 

virus obtained from peony and narcissus plants in New Zealand. Plant Disease 

Reporter, 62(11): 925–928. 

13. Loebenstein G., Lawson R. H., Brunt A. A. (eds.), 1995. Virus and Virus-like 

Diseases of Bulb and Flower Crops. Wiley, Jerusalem. 

14. Ningsheng W., Yunfeng W., 1990. About tobacco rattle virus peony isolate (TRV- 

Pa). Acta Phytopathology Sinica, 20(4): 247–251. 

15. Robinson D. J., Harrison B. D. 1989. Tobacco rattle virus. AAB Descriptions of 

Plant Viruses, 346 (No 12 revised). 6. 

16. Samuitienė M., Navalinskienė M. 1999. Identification of peony ringspot causal 

agent and its distribution in Lithuania. Material of International Scientific 

Conference on Plant Genefund Accumulation, Evaluation and Protection in the 

Botanical Gardens. 1–2 July, Vilnius, 85–87. 

17. Samuitienė M., Navalinskienė M., 2000. Natural occurrence of Tobacco rattle 

tobravirus on ornamental plants in Lithuania. Biologija, 2: 293–295. 

18. Spiegel S., Loebenstein G., 1995. Control of virus diseases. In: Loebenstein G., 

Lawson R. H., Brunt A. A. (eds.). Virus and Virus-like Diseases of Bulb and 

Flower Crops. Wiley, Jerusalem: 203–218. 

19. Weidemann H. -L. 1995. Detection of tobacco rattle virus in potato tubers and 

roots by polymerase chain reaction (PCR). Journal of Phytopathology, 143: 

455–458. 

20. Макутенайте М. 1974. Вирусные болезни декоративных растений семейства 

Ranunculaceae в Литве. Труды ВНИИ защиты растений, 4: 80–82. 

207


Tabako garbanotosios dryžligės (Tobacco rattle virus, TRV) 

viruso infekcijos bijūnuose (Paeonia L.) tyrimas 

M. Samuitienė, M. Navalinskienė, S. Dapkūnienė 

Santrauka 

Darbo tikslas buvo įvertinti mūsų šalies botanikos sodų kolekcijose auginamų lietuviškos 

selekcijos ir užsieninių veislių bijūnų pažeistumą bijūnų žiediškąja dėmėtlige ir identifikuoti 

šios virusinės ligos sukėlėją. Tikrinant Lietuvoje auginamų dekoratyvinių augalų 

kolekcijų fitosanitarinę būklę, buvo aptinkama pavienių bijūnų augalų su ryškiais bijūnų 

žiediškosios dėmėtligės požymiais. Sergančių augalų buvo aptinkama panašiu dažnumu tarp 

Paeonia lactiflora, P. lutea, P. officinalis, P. suffruticosa rūšių ir užsieninių, ir lietuviškos 

selekcijos veislių augalų. Buvo surinkti sergančių augalų pavyzdžiai virusinės ligos sukėlėjo 

tyrimams. Iš pažeistų augalų išskirtas ir klasikiniais virusologijos (augalų indikatorių, 

elektroninės mikroskopijos) bei moderniais molekulinės biologijos (DAS-ELISA, AT-PGR) 

metodais identifikuotas tabako garbanotosios dryžligės (Tobacco rattle virus, TRV) virusas. 

Reikšminiai žodžiai: DAS-ELISA, RT-PCR, tirti augalai, Tobacco rattle virus. 

208




Evaluation of agronomical characters and 

resistance to fungal diseases of apple cultivars 

Audrius Sasnauskas, Dalia Gelvonauskienė 


Lithuania, e-mail A.Sasnauskas@lsdi.lt 

In 2004–2009 at the Lithuanian Institute of Horticulture blooming period and abundance, 

trunk cross sectional area, yield, productivity, resistance to scab (Venturia inaequalis) and 

apple blotch (Phyllosticta mali) were studied in 7 apple (Malus × domestica Borkh.) cultivars. 

Trees were grafted on rootstock B.118 with spacing 4 × 2.5 m. 

‘Auksis’ and ‘Connell Red’ were the best ones among the tested cultivars. The following 

apple cultivars were distinguished for particular characteristics: ‘Kim’ – earliest blooming, 

‘Winterbanana’ – latest blooming, ‘Connell Red’– blooming abundance, ‘Connell Red’ – 

weakest growth, ‘Auksis’ and ‘Connell Red’ – yield in young orchard, ‘Auksis’ and ‘Birgit 

Bonnier’– resistance to scab, ‘Auksis’ and ‘Connell Red’– resistance to apple blotch. 

Key words: apple, cultivars, growth, phenology, Phyllosticta mali, productivity, Venturia 

inaequalis, yield. 

Introduction. Superior fruit qualities, high productivity, resistance to diseases 

and pests, winter hardiness, storability and marketability are the crucial requirements 

to new apple cultivars (Kellerhals et al., 1999; Kellerhals et al., 2003; Sansavini et al., 

2005; Rutkowski et al., 2005). Apple scab (Venturia inaequalis) is one of the most 

important diseases in fruit orchards in Lithuania (Raudonis, Valiuškaitė, 2003). High 

quality scab resistant cultivars have been developed over the last 50 years. Scab resistance 

determined by the gene V f 

, derived from Malus floribunda 821, has been 

considered durable for a long time (Kellerhals et al., 1999). However, before the appearance 

of races of scab virulent to V f 

(Parisi et al., 1993) it was obvious that breeding 

for scab resistance should include alternative sources of resistances. On the other 

hand, cultivars with durable resistance to other diseases – apple blotch (Phyllosticta 

mali), powdery mildew (Podosphaera leucotricha) and fire blight (Erwinia amylovora) 

– are important for ecological and economical considerations. Resistance to 

biotic and adaptation to abiotic factors of newly introduced apple cultivars and promising 

hybrids is being studied at the Lithuanian Institute of Horticulture (Sasnauskas 

et al., 2006; Gelvonauskienė et al., 2007; Sasnauskas et al., 2007; Sasnauskas et 

al., 2009). 

Objective of the work was to study some agronomical characters and resistance 

to fungal disease of the introduced apple cultivars. 

209

Object, methods and conditions. T r i a l y e a r s a n d p l a c e. The trial of 7 

apple cultivars was planted at the Lithuanian Institute of Horticulture in the spring of 

2004. Trees were grafted on B.118 rootstock. Evaluation and characterization of the 

cultivars was performed in 2006–2009. 

M e t e o r o l o g i c a l c o n d i t i o n s. The late spring frost at the beginning of 

bloom injured blossoms in 2007. At this time the minimal air temperature above the 

ground dropped from -3 °C to -2.5 °C, what injured fruit settings. Such conditions 

negatively affected yield parameters. In 2006 during flowering and after it weather 

was cold, but were no frosts and such condition didn’t affect yield. 

P l a n t m a t e r i a l. The following introduced apple cultivars were compared 

with the standard cv. ‘Auksis’ (Lithuania): ‘Bavendorf’ (Germany), ‘Birgit 

Bonnier’ (Sweden), ‘Connell Red’ (USA), ‘Kim’ (USA), ‘Nyckelby’ (Sweden) and 

‘Winterbanana’ (USA). 

E x p e r i m e n t a l d e s i g n. The trees were planted at the distance of 4 × 2.5 m. 

The trial was established in five replications. Each plot contained 1 fruit-tree. They were 

formed as spindle. Growing, fertilizing, pest, disease and weed control, soil cultivation, 

pruning, shaping and care of apple cultivars were maintained as recommended for 

commercial orchards (Intensyvios obelų ir kriaušių auginimo technologijos, 2005). 

O b s e r v a t i o n s a n d s t a t i s t i c a l a n a l y s i s. In the trial the following 

characters of apple cultivars was established: blooming periods, days; blooming 

abundance, scores (1-no blooming detected on trees, 9-bloomed more than 90 % of 

flowers); apple tree trunk cross-section area, cm 2 ; yield, t ha -1 ; productivity, kg/cm 2 , 

resistance to scab (Venturia inaequalis) and apple blotch (Phyllosticta mali), scores 

(1-no disease symptoms detected on leaves, 9-injured more than 75 % of leaf area). All 

data were subjected to analysis of variance. The significance of differences between the 

cultivars was estimated at 0.05 levels (Fisher’s Protected LSD and Duncan’s Multiple 

Range Test) (Tarakanovas, Raudonius, 2003). 

Results. B l o o m i n g p e r i o d. Apple trees started blooming in their third 

year in the orchard. All the blooming stages (beginning of blooming, beginning of 

full blooming, end of full blooming and end of blooming) began earliest in ‘Kim’, 

while the latest blooming was observed in ‘Winterbanana’ trees (Table 1). According 

to investigation data, blooming period continued 10–12 days. 

Table 1. Dates of blooming periods of apple cultivars 

1 lentelė. Obelų veislių žydėjimo tarpsniai 

Babtai, 2006–2008 

Cultivars 

Veislės 

Beginning of 

blooming 

(month, day) 

Žydėjimo pradžia, 

mėn., diena 

Beginning of full 

blooming 

(month, day) 

Masinio žydėjimo 

pradžia, mėn., diena 

210 

End of full blooming 

(month, day) 

Masinio žydėjimo 

pabaiga, 

mėn., diena 

End of blooming 

(month, day) 

Žydėjimo pabaiga, 

mėn., diena 

1 2 3 4 5 

‘Auksis’ 05-12 ab* 05-16 ab 05-20 ab 05-22 a 

‘Bavendorf’ 05-16 bcd 05- 20 bcd 05-23 bcd 05-26 bc 

‘Birgit Bonnier’ 05-16 bcd 05-20 bcd 05-22 bcd 05-25 abc



1 2 3 4 5 

‘Connell Red’ 05-17 cd 05-19 bcd 05-24 bcd 05-27 bc 

‘Kim’ 05-10 a 05-15 a 05-18 a 05-21 a 

‘Nyckelby’ 05-18 cd 05-22 cd 05-26 d 05-28 bc 

‘Winterbanana’ 05-18 d 05-23 d 05-25 cd 05-29 c 

Mean 

05-16 05-19 05-22 05-25 

Veislių vidurkis 

* Duncan’s multiple range t-test / Duomenys apskaičiuoti pagal Dunkano kriterijų Means 

followed by the same letter are not significantly different at P = 0.05 

* Tarp vidurkių, pažymėtų tomis pačiomis raidėmis, nėra esminių skirtumų, kai P = 0,05 

B l o o m i n g a b u n d a n c e. In the third year of growth apple trees ‘Connell 

Red’ (5.5 scores) bloomed most abundantly, while cvs. ‘Kim’ (1.1 scores) and ‘Auksis’ 

(1.3 scores) bloomed little (Table 2). In the fourth year of growth apple trees bloomed 

little (3.3 scores). During the evaluation period spring frosts in this year at the beginning 

of bloom injured blossoms. In the fifth year of growth apple trees bloomed better 

(4.5 scores). Apple trees ‘Connell Red’ (6 scores) and ‘Nyckelby’ (5.3 scores) bloomed 

most abundantly. On the other hand, cv. ‘Bavendorf’ (2.3 scores) bloomed little. 

Table 2. Blooming abundance of apple cultivars 

2 lentelė. Obelų žydėjimo gausumas 

Cultivars 

Veislės 

Babtai, 2006–2008 

Blooming abundance (scores) 

Obelų žydėjimo gausumas, balais 

2006 2007 2008 

‘Auksis’ 1.3 a* 4 bcd 4.6 ab 

‘Bavendorf’ 1.5 ab 2 a 2.3 a 

‘Birgit Bonnier’ 2.5 ab 3 ab 3.6 ab 

‘Connell Red’ 5.5 d 6 d 6 c 

‘Kim’ 1.1 a 3 ab 5 bc 

‘Nyckelby’ 2.5 ab 2 a 5.3 bc 

‘Winterbanana’ 3.5 bcd 5.6 cd 4 ab 

Mean 

2.5 3.3 4.5 


* Duncan’s multiple range t-test / Means followed by the same letter are not significantly 

different at P = 0.05 

* Duomenys apskaičiuoti pagal Dunkano kriterijų / Tarp vidurkių, pažymėtų tomis pačiomis 

raidėmis, nėra esminių skirtumų, kai P = 0,05 

T r e e g r o w t h. In the fifth year of growth apple trees of the investigated cultivars 

showed that the trees of ‘Birgit Bonnier’ (36.7 cm 2 ) and ‘Kim’ (36.5 cm 2 ) grew 

significantly stronger, while ‘Connell Red’ (23.5 cm 2 ) – weaker, compared with the 

other tested apple cultivars (Fig. 1). 

211

Fig. 1. Trunk cross sectional area (cm 2 ) 

1 pav. Obelų veislių kamieno skerspjūvio plotas, cm 2 

Babtai, 2008 

Y i e l d a n d p r o d u c t i v i t y. The yield per year is cumulatively presented in 

Fig. 2. These data show that the highest cumulative yield per hectare is from ‘Auksis’ 

(23.7 t ha -1 ). ‘Connell Red’ (21.8 t ha -1 ) gives almost the same yield. The next in yield 

capacity were cvs. ‘Bavendorf’ (18 t ha -1 ) and ‘Nyckelby’ (15.1 t ha -1 ). ‘Kim’ (6.1 t ha -1 ), 

‘Birgit Bonnier’ (8.9 t ha -1 ) and ‘Winterbanana’ (13.3 t ha -1 ) had the lowest yield. 

Fig. 2. Cumulative yield of apple cultivars (t ha -1 ) 

2 pav. Obelų veislių suminis derlius, t ha -1 

Babtai, 2006–2008 

212

The average of productivity of the investigated apple cultivars was 0.40 kg/cm 2 

(Fig. 3). The highest productivity was assessed for cultivar ‘Auksis’ (0.77 kg/cm 2 ). 

The significantly lowest productivity showed ‘Birgit Bonnier’ (0.14 kg/cm 2 ) and ‘Kim’ 

(0.16 kg/cm 2 ). Productivity of cultivars ‘Bavendorf’ (0.42 kg/cm 2 ) ‘Winterbanana’ 

(0.42 kg/cm 2 ) and ‘Nyckelby’ (0.50 kg/cm 2 ) was close to the average and significantly 

differed from cultivars discussed before. 

Fig. 3. Productivity of apple cultivars (kg/cm 2 ) 

3 pav. Obelų veislių produktyvumas, kg/cm 2 

Babtai, 2008 

D i s e a s e r e s i s t a n c e. Tested apple genotypes demonstrated different field 

resistance to apple scab. In the fourth year in orchard, cvs. ‘Bavendorf’ (2.6 scores) 

and ‘Nyckelby’ (2.9 scores) were the most susceptible on leaves (Table 3). In the 

fifth year in orchard, ‘Kim’ (2 scores) was the most susceptible to this disease. In the 

sixth year in orchard, all cultivars displayed higher apple scab symptoms on leaves. 

More resistant to this disease on leaves was cvs. ‘Auksis’, ‘Birgit Bonnier’ and ‘Kim’ 

(2 scores), while ‘Nyckelby’ (5 scores) was the most susceptible to apple scab. 

All cultivars displayed apple blotch symptoms on leaves in 2007 (Table 3). 

Cv. ‘Auksis’ (1.1 scores) showed high resistance. Cv. ‘Bavendorf’ (2.6 scores) was 

the most susceptible. In the fifth and sixth year in orchard, cvs. ‘Auksis’ and ‘Connell 

Red’ (1 score) were free from apple blotch symptoms on leaves. On the other hand, 

cv. ‘Kim’ and ‘Birgit Bonnier’ were the most susceptible to this disease. 

213

Table 3. Scab and apple blotch injury on apple-tree (scores) 

3 lentelė. Obelų pažeidimas rauplėmis ir filostiktoze, balais 

Scab 

Rauplės 

Apple blotch 

Filostiktozė 

Babtai, 2007–2009 

Cultivars 

Veislės 

2007 2008 2009 

Vidurkis 

Average 

2007 2008 2009 

Vidurkis 

Average 

‘Auksis’ 1.7 1.6 2.0 1.7 1.1 1.0 1.0 1.03 

‘Bavendorf’ 2.6 1.5 3.0 2.37 2.6 1.0 2.3 1.97 

‘Birgit Bonnier’ 1.3 1.9 2.0 1.73 2 1.5 2.0 1.87 

‘Connell Red’ 1.8 1.8 3.2 2.27 1.8 1.0 1.0 1.27 

‘Kim’ 1.8 2.0 2.0 1.93 1.6 1.5 2.6 1.9 

‘Nyckelby’ 2.9 1.6 5.0 3.17 1.5 1.0 2.0 1.5 

‘Winterbanana’ 1.4 1.8 2.2 1.80 1.8 1.7 1.8 1.77 

Average 1.92 1.74 2.78 2.15 1.77 1.24 1.82 1.61 


LSD 05 

/ R 05 

0.49 0.26 1.10 0.51 0.38 0.13 0.51 0.36 

Discussion. One of the requirements for successful pollination is that the pollinator 

must flower at the same time as the cultivar to be pollinated (Grauslund, 1996). For 

this purpose cultivars were divided into groups of similar blooming time. The early 

blooming apple cultivars suffer a great risk of being damaged by the late spring frost 

(Kiprijanovski et al., 2009). Results of investigation show that ‘Kim’ is characterized 

with the earliest start of blooming, while ‘Winterbanana’ start blooming much later. 

During three years of full blooming apple trees of cv. ‘Connell Red’ bloomed 

most abundantly. Less intensive bloom was characterized for other apple cultivars. 

Apple trees of cv. ‘Bavendorf’ bloomed worst. 

Trunk cross sectional area (TCSA) is an integral indicator of the development of 

the complete above ground part of the fruit-tree (Ristevski, 1986). The data of investigation 

showed that cvs. ‘Birgit Bonnier’ and ‘Kim’ had the largest TSCA at the end 

of the fifth year, whereas the ‘Connell Red’ type had the smallest. 

Highly significant yield differences were found among the trees of the tested apple 

cultivars. The highest yield and productivity showed trees of ‘Auksis’ and ‘Connell 

Red’. As reported earlier (Uselis et al., 2007; Sasnauskas et al., 2008), cvs. ‘Auksis’ 

and ‘Connell Red’ were the most productive. The lowest yield and productivity was 

observed for ‘Kim’ and ‘Birgit Bonnier’. 

Resistance breeding in apple is aimed at the development of cultivars durably 

resistant to biotic and abiotic stresses. Resistant cultivars are suited for sustainable 

and ecological production methods due to less application of pesticides needed for 

healthy growth. Tolerance to abiotic stresses can promote a better adaptation to the 

environment and climatic changes (Peil, 2009). Our investigation data demonstrated 

that ‘Auksis’ and ‘Birgit Bonnier’ were resistant to scab, ‘Auksis’ and ‘Connell Red’ – 

resistant to apple blotch. 

214

Conclusions. 1. ‘Auksis’ and ‘Connell Red’ were the best ones among the tested 

cultivars. 

2. The following apple cultivars were distinguished for particular characteristics: 

‘Kim’ – earliest blooming, ‘Winterbanana’ – latest blooming, ‘Connell Red’ – blooming 

abundance, ‘Connell Red’ – weakest growth, ‘Auksis’ and ‘Connell Red’ – yield 

in young orchard, ‘Auksis’ and ‘Birgit Bonnier’– resistance to scab, ‘Auksis’ and 

‘Connell Red’ – resistance to apple blotch. 

Gauta 2009 07 01 


References 

1. Gelvonauskienė D., Sasnauskas A., Gelvonauskis B. 2007. The breeding of apple 

tree resistant to European canker (Nectria Galligena Bres.). Sodininkystė ir 

daržininkystė, 26 (3): 174–178. 

2. Grauslund J. 1996. Flowering dates of pome and stone fruit cultivars –10 years 

results. Acta Horticulturae, 423: 31–37. 

3. Intensyvios obelų ir kriaušių auginimo technologijos. 2005. N. Uselis (sudaryt.). 

Lietuvos sodininkystės ir daržininkystės institutas, Babtai. 

4. Kellerhals M., Viviani A., Goerre M., Gessler C. 1999. New challengers for 

apple breeding. Acta Horticulturae, 484: 131–134. 

5. Kellerhals M., Sauer C., Frey J., Hohn E. 2003. High fruit quality, optimal fruit 

set and durable disease: are these the requirements for new apple varieties 

Proceedings Eufrin workshop on fruit quality, Bologna, Italy, 11–14 June. 

6. Kiprijanovski M., Arsov T., Gjamovski V., Damovski. 2009. Study of certain 

introduced apple cultivars in the Prespa region. Acta Horticulturae, 825: 

125–132. 

7. Parisi L., Lespinasse Y., Guillaumes J., Krüger J. 1993. A new race of Venturia 

inaequalis virulent to apples with resistance due to the Vf gene. Phytopathology, 

83: 533–537. 

8. Peil, A., Duneman F., Flachowsky H., Hanke M.V. Update on breeding for disease 

resistance in apple. Proceedings of the COST Action 864: Combining traditional 

and advanced strategies for plant protection in pome fruit growing. 3–5 

June, Valencia, Spain, 22. 

9. Raudonis L., Valiuškaitė A. 2003. Research on pest and disease control in horticultural 

plants and its development in Lithuania. Sodininkystė ir daržininkystė, 

22(3): 3–14. 

10. Ristevski B. 1986. Opsto ovostarstvo. Nasa kniga, Skopje. 

11. Rutkowski K. P., Kruczynska D. E., Plocharski W., Wawrzynczak A. 2005. 

Scab resistant apple cultivars – quality and storage. Acta Horticulturae, 682: 

681–686. 

215

12. Sansavini S., Belfanti E., Costa F., Donati, F. 2005. European apple breeding 

programs turn to biotechnology. Chronica Horticulturae, 45(2): 16–19. 

13. Sasnauskas A., Gelvonauskienė D., Gelvonauskis B., Bendokas V., Baniulis D. 

2006. Resistance to fungal diseases of apple cultivars and hybrids in Lithuania. 

Agronomy Research, 4: 349–352. 

14. Sasnauskas A., Gelvonauskienė D., Bendokas V., Stanys V., Rugienius R., 

Bobinas Č., Baniulis D. 2007. Characterization of scab resistant Lithuanian apple 

cultivars. Acta Horticulturae, 760: 507–511. 

15. Sasnauskas A., Gelvonauskienė D., Viškelis P., Šabajevienė G., Duchovskis P. 

2008. Introdukuotų obelų veislių tyrimų apžvalga. Sodininkystė ir daržininkystė, 

27(3): 77–84. 

16. Sasnauskas A., Gelvonauskienė D., Šikšnianienė J. B., Šabajevienė G., 

Duchovskis P., Bobinas Č. 2009. Investigation of biological traits of fifteen apple 

cultivars. Acta Horticulturae, 825: 97–102. 


analizė taikant kompiuterines programas ANOVA, STAT, SPILT-PLOT iš paketo 

SELEKCIJA ir IRRISTAT. Metodinė priemonė. Akademija: 57. 

18. Uselis N., Duchovskis P., Kviklys D., Šabajavienė G. 2007. Effect of planting 

systems and canopy form on apple trees grafted on P 22. Sodininkystė ir daržininkystė, 

26(4): 22–29. 


Obelų veislių agronominių požymių ir atsparumo 

grybinėms ligoms tyrimas 

A. Sasnauskas, D. Gelvonauskienė 

Santrauka 

2004–2009 m. Lietuvos sodininkystės ir daržininkystės institute tirta septynių introdukuotų 

obelų (Malus × domestica Borkh.) veislių žydėjimo tarpsniai, žydėjimo gausumas, vaismedžių 

augumas, derlius, produktyvumas, atsparumas rauplėms (Venturia inaequalis) ir filostiktozei 

(Phyllosticta mali). Dvimečiai obelų sodinukai su B.118 poskiepiu pasodinti 2004 m. pavasarį. 

Sodinimo schema – 4 × 2,5 m, po vieną vaismedį laukelyje penkiais pakartojimais. 

Pagal tirtų požymių visumą, vaismedžiai ‘Auksis’ ir ‘Connell Red’ įvertinti geriausiai. 

Tirtais požymiais išsiskyrė šios obelų veislės: ‘Kim’ – ankstyvu žydėjimu, ‘Winterbanana’ – 

vėlyvu žydėjimu, ‘Connell Red’ – žydėjimo gausumu, ‘Connell Red’ – silpniausiu augumu, 

‘Auksis’ ir ‘Connell Red’ – derliumi jauname sode, ‘Auksis’ ir ‘Birgit Bonnier’– atsparumu 

rauplėms, ‘Auksis’ ir ‘Connell Red’ – atsparumu filostoktozei. 

Reikšminiai žodžiai: augumas, derlius, fenologija, obelys, Phyllosticta mali, produktyvumas, 

veislės, Venturia inaequalis. 

216




Influence of preplant and vegetable crop rotation links 

on carrot yield and damage of pests 

Roma Starkutė, Laisvūnė Duchovskienė, Vytautas Zalatorius 


Lithuania, e-mail r.starkute@lsdi.lt 

At the Lithuanian Institute of Horticulture in 2003–2007 there were investigated the 

most suitable plants for green manure and evaluated their influence on ecologically grown 

carrot yield and damage of pests. Experiments were carried out in the experimental field for 

ecological vegetable growing, in calcaric epihypogleyic luvisol of sandy loam on light loam. It 

was established that biomass of the plants grown for green manure left in the ploughing layer 

uneven amount of organic matter. Pea and oat mixture produced the biggest amount of organic 

matter (43.2 t ha -1 ), barley – the least one (24.5 t ha -1 ). All the sideral plants influenced humus 

positively. All the preplants increased carrot yield. The biggest carrot yield (correspondingly 

40.4 and 41.2 t ha -1 ) was obtained growing them after barley and pea-oat mixture for green 

manure. In the first year after harvesting of the plants for green manure, when carrot yield 

was gathered, there was found only very small amount of root-crops damaged by pests (Psila 

rosae Fabr. and Pemphigus phenax Born et Blunck). At the end of rotation, i. e. in third year 

after plant ploughing for green manure, the percent of carrot root-crops damaged by pests 

increased. The least amount of damaged root-crops was found in carrot, which preplant in 

the beginning of rotation was barley for green manure. 

Key words: carrot root-crops, crop rotation, green manure, marketable yield, Pemphigus 

phenax, Psila rosae. 

Introduction. Carrot is widely grown vegetable in Lithuania especially valued 

because of carotene in its content (Gaučienė, 2001; Gaučienė, Viškelis, 2001). It is 

rather difficult to grow carrot ecologically, since they are demanding to nutrients, 

besides, they are attacked by diseases and pests. The most prevalent carrot pests are 

carrot flies, aphids and bugs. 

The suitable selection of preplants and creation of crop rotations is one of the most 

important means to supply nutrients to ecologically grown plants, to destroy weeds 

and to protect plants from diseases and pests. Soil preparation and sowing time, the 

ratio of humidity and nutrients in the soil, weediness, pest prevalence in the crop and 

the degree of infection by diseases depend on preplant (Danilčenko et al., 2004; Luik, 

1997; Lampkin, 1990). 

Sideral plants for green manure play important role in the crop rotation of 

217

ecological vegetable growing. In recent years they became especially popular, because 

only little amount of manure is accumulated in farms. Besides, green manure is much 

more cheaper and its influence in the first year often is better than this of manure 

(Алексеев, 1996; Arlauskienė, Maikštėnienė, 2002). For green manure, leguminous 

plants are sown most often (Abiodun et al., 2008; Šarūnaitė et al., 2008), but as the 

experience of foreign countries show, cereals are good as well (Andersson, Wivstad, 

1992). Various plant mixtures are especially suitable. After the ploughing of the plants 

for green manure, the soil is enriched by much organic matter, biological soil properties 

improve, and because of the phytoncides present in crucials phytosanitaric condition 

of the soil improves also (Žekonienė, 2002; Pretty, 1995). It is recommended to select 

for carrot in crop rotation early preplants, in order soil structure would be regenerated. 

Otherwise, there are good conditions for carrot fly development (Lampkin, 1990). 

The aim of the study is to select the most suitable plants for green manure and 

to establish their influence on ecologically grown carrot yield and pest prevalence in 

the crop. 

Object, methods and conditions. Experiments were carried out at the Lithuanian 

Institute of Horticulture, in the experimental field for ecological vegetable 

growing in 2003–2007. Soil – calcaric epihypogleyic luvisol of sandy loam on light 

loam (IDg 8-k, / Calc(ar)i –Epihypogleyc Luvisols LVg-p-w-cc) (Buivydaitė et al., 

2001). pH KCL 

of ploughing layer – 7.4, agile phosphorus (P 2 

O 5 

) – 211–370 mg kg -1 , 

agile potassium (K 2 

O) – 151–249 mg kg -1 , humus 1.64 – 1.69 %, mineral nitrogen at 

a depth of 0–60 cm – 92 kg ha -1 . 

Two field experiments were carried out according to the same schema; one of 

them was started in 2003, the other – in 2004. 

In the first year, for green manure there was grown barley, barley with clover 

undercrop (of the first year), summer wheat, and peat-oat mixture. Black fallow was 

regarded as control. In the second year, after each plant for green manure there were 

grown vegetables of four types – carrot, red beet, onion, and cabbage. Plants were 

grown according to the s c h e m e: 

First yield – Second year – Third year – Fourth year 

Various plants for green manure – Cabbage – C a r r o t – Red beet 

Various plants for green manure – C a r r o t – Red beet – Onion 

Various plants for green manure – Red beet – Onion – Cabbage 

Various plants for green manure – Onion – Cabbage – C a r r o t. 

Investigations were carried out in four replications. Area of sideral variants (of all 

the plots) was 264 m 2 . Area of vegetable record plots – 14 m 2 . Preplant of the plants 

for green manure – black fallow. 

Early in the spring, when the soil is already dry, it is cultivated and harrowed. In the 

second decade of May for green manure there were sown summer wheat (180 kg ha -1 ), 

peat and oat mixture (125 : 125 kg ha -1 ) and barley (170 kg ha -1 ). When they germinated, 

half of barley field was sown with clover undercrop (14 kg ha -1 ). Seed rate was 

calculated in 100 % economical value. 

Pea and oat mixture was cut during mass blooming, summer wheat and barley – 

during spikes forming. Grass was chopped fine and ploughed up. Straw was raked 

218

from the plot, in which clove undercrop was sown, and late in the autumn clover was 

ploughed down. Black fallow was cultivated according to layer method: twice ploughed, 

three times cultivated. In order to establish plant fresh weight yield, in 10 m 2 area of 

every plot plants were cut and weighted. For determination of dry matter and yield 

and chemical analyses, the sample of 1 kg fresh weight was taken. 

Pest damages to carrot root-crops were evaluated during harvesting: the injured 

carrots were selected, weighted and the percent of damage from the total yield was 

established. During vegetation, carrot crop was observed according to the standard 

methods of disease and pest record (Žemės ūkio augalų kenkėjai, ligos ir jų apskaita, 

2002). Experimental data was statistically processed by ANOVA program (Tarakanovas, 

Raudonius, 2003). 

M e t e o r o l o g i c a l c o n d i t i o n s. The spring of 2003 was dry, but warmer 

than the multiannual average. Growth conditions for plants designed for green manure 

were average. Summer time and September were slightly warmer, but the amount of 

precipitation was close to the multiannual average. In 2004 air was cooler and more 

humid than the multiannual data show, but the conditions for carrot growing were 

favourable. In May and June of 2005, there was more precipitation than on the average. 

July was hot and dry. August was much more humid than usually. Conditions 

for carrot germination, growth and maturing were average. The summer of 2006 was 

hot and very dry. Average air temperature was almost 1 °C higher than the multiannual 

average, and precipitation fell 20.5 mm less than the multiannual average. July 

and August were especially dry and hot; September was warm in comparison with the 

multiannual average, but humid (there was 1.5 more precipitation than the multiannual 

average). In 2007 vegetation period was cool. Air temperature in April–October was 

1.2 °C lower than the multiannual average, and precipitation fell 12.8 mm more than 

the multiannual average. 

Results. According to the average data of two years, pea-oat mixture produced the 

biggest amount (43.2 t ha -1 ) of fresh weight, and barley – the smallest one (24.5 t ha -1 ) 

(Table). Their fresh weight in comparison with this of pea-oat mixture decreased 

18.7 t ha -1 . The biggest amount of dry matter (7.3 t ha -1 ) was ploughed down with 

summer wheat fresh weight. The amount of barley, barley with clover undercrop, 

and pea-oat mixture dry matter was correspondingly smaller 1.8, 1.4, 0.8 t ha -1 . The 

calculation of the amount of nutrients ploughed down into soil together with fresh 

weight showed that 104 kg ha -1 of nitrogen got into soil together with pea-oat fresh 

weight. This is 42 kg ha -1 more than after barley, 17 kg ha -1 more than after barley 

with clover undercrop, 6 kg ha -1 more than after summer wheat. The biggest amount 

of phosphorus (21.0 kg ha -1 ) got into soil when barley with clove undercrop fresh 

weight was ploughed down, the least one (14 kg ha -1 ) – when barley fresh weight was 

ploughed down. The biggest amount of potassium (150 kg ha -1 ) with fresh manure 

was introduced by ploughing down pea-oat mixture, the least amount (79 kg ha -1 ) – 

by ploughing down barley. 

219

Table. Amount of nutrients in over-ground mass of sideral plants 

Lentelė. Maisto medžiagų kiekis sideratinių augalų antžeminėje masėje 

Variants 

Variantai 

Babtai, 2003–2004, average data of both fields 

Babtai, 2003–2004 m., vidutiniai abiejų laukų duomenys 

Organic manure 

Organinės trąšos (t ha -1 ) 

natural expression 

natūralioji 

išraiška 

220 

dry matter 

sausosios 

medžiagos 

Nutrients 

Maisto medžiagos (t ha -1 ) 

N P 2 

O 5 

K 2 

O 

Black fallow (control) 

Juodasis pūdymas (kontrolė) 

Barley for green manure 

24.5 4.1 62 14 79 

Miežiai žaliajai trąšai 

Barley with clover undercrop for 30.2 5.9 87 21 121 

green manure 

Miežiai su dobilų įsėliu žaliajai trąšai 

Summer wheat for green manure 32.5 7.3 98 19 125 

Vasariniai kviečiai žaliajai trąšai 

Pea-oat mixture for green manure 43.2 6.5 104 20 150 

Žirnių-avižų mišinys žaliajai trąšai 

LSD 05 

/ R 05 

3.0 0.5 

In the first year after plant gathering for green manure (in I field – in 2004, in II 

field – in 2005) all the preplants increased yield. Carrot marketable yields in comparison 

with the yields obtained growing carrot after black fallow increased on the average 

12.5 t ha -1 . The biggest carrot yields (correspondingly 40.4 and 41.2 t ha -1 ) were obtained 

growing them after barley and pea-oat mixture for green manure (Fig. 1). 

Carrot marketable yields in comparison with the yields obtained growing carrot 

in black fallow field increased 14.2 and 14.9 t ha -1 . Summer wheat and pea with clove 

undercrop for green manure also significantly (correspondingly 11.4 and 9.4 t ha -1 ) 

increased root-crop yield. 

Barley and barley with clove undercrop were the best preplants for carrot grown 

after cabbage (in the second year after green manure ploughing up, in I field – in 

2005, in II field – in 2006). Carrot marketable yields in comparison with the yields 

obtained growing carrot after cabbage in black fallow field increased 7.1 and 6.2 t ha -1 . 

Summer wheat for green manure also 2.2 t ha -1 increased marketable root-crop yield. 

Nevertheless, growing carrot after cabbage, which preplant was pea-oat mixture, there 

was obtained 0.8 t ha -1 smaller yield in comparison with the yield obtained growing 

carrot after cabbage in black fallow field. 

In the third year after plant gathering for green manure (in I field – in 2006, in 

II field – in 2007), when carrot was grown after cabbage and onion, the tendency of 

carrot yield decrease was observed in comparison with the yields obtained in the first 

and second years of growing. 

The most durable preplants were barley and summer wheat. Carrot marketable 

yield, growing them after cabbage and onion, increased correspondingly 5.8 and 

5.4 t ha -1 , in comparison with the yield, which produced carrot grown in black fallow 

field.

Fig. 1. Influence of preplants and crop rotation links on carrot root-crop marketable yield 

1 pav. Priešsėlių ir sėjomainos grandžių įtaka morkų šakniavaisių prekiniam derliui 

Babtai, average data of 2004–2006 and 2005–2007 / 

vidutiniai 2004–2006 ir 2005–2007 m. duomenys 

In the first year after plant gathering for green manure, when carrot was harvested, 

there was found very small amount of root-crops damaged by pests. Only 0.2 % of 

damaged root-crops were found in carrot grown after black fallow and 0.1 % – after 

barley for green manure. 

Fig. 2. Influence of preplants on pest (P. rosae Fabr. and 

P. phenanax Born et Bunck) prevalence in carrot 

2 pav. Priešsėlių įtaka kenkėjų (P. rosae Fabr. ir 

P. phenanax Born et Bunck) išplitimui morkose 

Babtai, average data of 2004–2006 and 2005–2007 / 

vidutiniai 2004–2006 ir 2005–2007 m. duomenys 

221

In the second year after plant gathering for green manure, on the average the 

biggest amount (1.8 %) of damaged root-crops in comparison with control was found 

in carrot grown after cabbage, which preplant – pea-oat mixture. On the average the 

least amount (correspondingly 0.5–0.6 %) of damaged root-crops in comparison with 

control was found in carrot grown after cabbage, which preplant – barley and barley 

with clover undercrop. 

In the third year after plant gathering for green manure, the amount of damaged 

root-crops in all the plots was similar and fluctuated from 1.2 up to 1.3 %. Nevertheless, 

in carrot, which preplant in the beginning of crop rotation was barley ploughed up for 

green manure, there were only 0.6 % of damaged carrot. 

In the third year after plant gathering for green manure the percent of damaged 

carrot root-crops increased and marketable root-crop yield decreased. 

Discussion. In Integrated Plant Protection (IPP), cultural measures like preplants 

and plant rotation play very important role; especially it is important in ecological 

farming (Glosary et al., 1997). While growing carrot ecologically, green manure is 

one of the means to increase the amount of nutrients in the soil and to improve vegetable 

productivity. In crop rotation, when plants are being fertilized with green manure, 

much nitrogen, phosphorus and potassium is left in the soil (Kozlova, Kaminski, 

1998), but different plants assigned for green manure differently enrich the soil with 

organic matter. The biggest amount of fresh weight produced red cloves (Šlepetienė, 

Kinderienė, 2007). The same tendency was observed in the investigations carried out 

at the LIH. 

Scientists (Thorup-Kristensen, Boogaar, 1999; Rosa, Jabłońska-Ceglarek, 2008) 

established that all the plants for green manure increased carrot root-crop and other 

vegetable yield, but it was difficult to control carrot pests. Organic growers have had to 

rely on techniques such as covering the crop with a fine-mesh netting and manipulating 

the time of sowing and harvest (Ellis et al., 1987; Jönsson, 1992). The former method 

is costly, and conditions under the netting can be favourable to the development of 

fungal pathogens and weeds (Eichin et al., 1987; Peacock, 1991). A delay in sowing 

or premature harvesting both may result in yield loses (Ellis et al., 1987). In our study 

effect of preplants used for green manure detectible also after three years. After one-year 

effect of preplants for reducing damage of pests was highest, maybe field was clean 

from pupa of carrot fly. The preplant barley reduced the harm of carrot pests during 

all the years and standard yield losses were the smallest. According to Rämert (1996 

b), growing vegetables ecologically predators play very important role. We suggest 

that in treatment with barley predators inhibited pests. 

Conclusions. 1. Pea and oat mixture produced the biggest amount of fresh 

weight (43.2 t ha -1 ), barley – the least one (24.5 t ha -1 ). 

2. All the sideral plants influenced the positive changes in humus. 

3. All the preplants increased carrot yield. The biggest carrot yields (correspondingly 

40.4 and 41.2 t ha -1 ) were obtained growing them after barley and pea-oat 

mixture for green manure. 

222

4. In the first year after plant gathering for green manure, when carrot was harvested, 

there was found very small amount of root-crops damaged by pests (P. rosae 

and P. phenax). At the end of crop rotation, i. e. in the third year, after plant gathering 

for green manure the percent of pest damaged carrot root-crops increased. The least 

amount of damaged root-crops was found in carrot, which preplant in the beginning 

of crop rotation was barley for green manure. 

Gauta 2009 06 30 


References 

1. Abiodun A. Kintomo a , Henry A. Akintoye a , Kayode O. Alasiri a ., 2008. 

Communications in Soil Science and Plant Analysis, 39(9 & 10): 1 261–1 268. 

2. Andersson T., Wivstad M. 1992. Vallen iväxtfö ijden: Betydelsen av vallensalder 

och botaniskasammansältning. – Uppsala, 38: 43 

3. Arlauskienė A., Maikštėnienė. 2002. Molingų dirvožemių savybių gerinimas 

ankštiniais augalais, jų biomasę panaudojant žaliajai trąšai. Žemdirbystė, 79(3): 

229–243. 

4. Danilčenko H. ir kt. 2004. Ekologinė daržininkystė. Kaunas, Akademija. 

5. Dufault C. P., Coaker T. H. 1987. Biology and control of the carrot rust fly, Psila 

rosae F. Agricultural Zoology Reviews, 2: 97–134. 

6. Eichin R., Deiser E., Buhl R. 1987. Netze und Vliese gegen Gemüsefliegen. 

Deutscher Gartenbau, 41: 206–213. 

7. Ellis P. R., Hardman J. A., Cole R. A., Phelps K. 1987. The complementary 

effects of plant resistance and choice of sowing and harvest times in reducing 

carrot fly (Psila rosae) damage to carrots. Annuals of Applied Biology, 111: 

415–424. 

8. Gaučienė O. 2001. Morkos. Babtai. 

9. Gaučienė O., Viškelis P. 2001. Tinkamiausių Lietuvoje auginti morkų (Daucus 

carota L.) derlius ir kokybė. Sodininkystė ir daržininkystė. 20(4)–1: 17–24. 

10. Jönsson B. 1992. Forecasting the timing of damage by the carrot fly. IOBC/ 

WPRS Bulletin, XVI 4: 40–43. 

11. Kozlova L., Kaminski J. R. 1998. Energy efficiency of crop rotations for sustainable 

agriculture. Agriculture, 64: 43–55 

12. Lampkin N. 1990. Organic farming. Farming press books, Ipswich, UK. 

13. Luik A. 1997. The influence of plants on insects. (In Estonian with English summary). 

Tartumaa kirjastus, Tartu. 

14. Oskam A. J., Vijftigschild R. A. N., Graveland C. 1997. Additional EU policy 

instruments for plant protection products. Final report. http://glossary.eea.europa.eu 

15. Peacock L. 1991. Effect on weed growth of short-term cover over organically 

grown carrots. Biological Agriculture and Horticulture, 7: 271–279. 

223

16. Pretty, J. N. 1995. Regenerating agriculture: policies and practice for sustainability 

and self – reliance. Earthscan, London. 

17. Rämert B. (a) 1996. Intercropping as a strategy for reducing damage to carrots 

caused by the carrot fly Psila rosae (F.). Biological Agriculture and Horticulture, 

13: 359–369. 

18. Rämert B. (b) 1996. The influence of intercropping and mulches on the occurrence 

of polyphagous predators in carrot fields in relation to carrot fly (Psila rosae 

(F.) (Dipt. Psilidae) damage. Journal of Applied Entomology, 120: 39–46. 

19. Rosa R., Jabłońska-Ceglarek R. 2008. The consecutive effect of using green 

manure forecrops and manurenin ‘BLIZZARD’ leek (Allium ampeloprasum ssp. 

porrum) cultivation, http://www.ejpau.media.pl/volume11/issue2/art-22.html 

20. Szwejda J., Wrzodak R. 2007. Phytophagous entomofauna occurring on carrot 

and plant protection methods. Vegetable Crops Research Bulletin, 67: 95–102. 

21. Šarūnaitė L., Kadžiulienė Ž., Kadžiulis L. 2008. Žemės ūkio mokslai. 15(1): 

10–16. 

22. Šlepetienė A., Kinderienė I. 2007. Humuso medžiagų pokyčiai kalvoto reljefo 

dirvožemyje praturtinus jį tarpinių augalų žalia mase. Žemdirbystė. 94(1): 

37–50. 


analizė, taikant kompiuterines programas ANOVA, STAT, SPLIT-PLOT iš paketo 

selekcija ir IRRISTAT. Akademija, Kauno r. 

24. Thorup-Kristensen K., Boogaar K. R. 1999.Vertical and horizontal development 

of the root system of carrots fallowing green manure. Plant and soil, 212(2): 

143–151(9). 

25. Žekonienė V. 2002. Žalioji trąša. Kaunas. 

26. Žemės ūkio augalų kenkėjai, ligos ir jų apskaita. 2002. J. Šurkus, I. Gaurilčikienė 

(sudaryt.). Lietuvos žemdirbystės institutas. Akademija, Kėdainių r. 

27. Алексеев С. В. П. 1996. Ппрактикум по экологии. М.: АО МДС. 


Priešsėlių ir daržovių sėjomainos grandžių įtaka morkų derliui ir 

kenkėjų daromiems pažeidimams 

R. Starkutė, L. Duchovskienė, V. Zalatorius 

Santrauka 

Lietuvos sodininkystės ir daržininkystės institute 2003–2007 metais ekologiškoms 

daržovėms auginti paruoštame bandymų lauke tirti žaliajai trąšai tinkamiausi augalai ir įvertinta 

jų įtaka ekologiškai auginamų morkų derliui bei kenkėjų daromiems pažeidimams. Nustatyta, kad 

žaliajai trąšai auginamų augalų biomasė armenyje paliko nevienodą organinės medžiagos kiekį. 

Daugiausia žaliosios masės (43,2 t ha -1 ) užaugino žirnių ir avižų mišinys. Vasariniai kviečiai – 

32,5 t ha -1 , miežiai su dobilų įsėliu – 30,2 t ha -1 . Mažiausiai žaliosios masės užaugino miežiai – 

24,5 t ha -1 . Humuso teigiamiems pokyčiams įtakos turėjo visi sideraliniai augalai. Visi priešsėliai 

224

didino morkų derlių. Didžiausias morkų derlius (atitinkamai po 40,4 ir 41,2 t ha -1 ) buvo auginant jas 

po miežių ir žirnių-avižų mišinio žaliajai trąšai. Pirmais po augalų nuėmimo žaliajai trąšai metais, 

nuėmus morkų derlių, kenkėjų (Psila rosae Fabr. ir Pemphigus phenax Born et Blunck) pažeistų 

šakniavaisių buvo rasta labai nedaug. Rotacijos pabaigoje, t. y. trečiais po augalų užarimo žaliajai 

trąšai metais, padidėjo kenkėjų pažeistų morkų šakniavaisių procentas. Mažiausiai pažeistų 

šakniavaisių rasta morkose, kurių priešsėlis rotacijos pradžioje buvo miežiai žaliajai trąšai. 

Reikšminiai žodžiai: morkų šakniavaisiai, Pemphigus phenax, prekinis derlius, Psila 

rosae, sėjomaina, žalioji trąša. 

225




Effect of essential oils on fungi isolated from 

apples and vegetables 

Elena Survilienė 1 , Alma Valiuškaitė 1 , Vilija Snieškienė 2 , 

Antanina Stankevičienė 2 

1 


Lithuania, e-mail: e.surviliene@lsdi.lt; a.valiuskaite@lsdi.lt 

2 

Kaunas Botanical Garden of Vytautas Magnus University; Z. E. Žilibero 6, 

Kaunas, Lithuania, e-mail: v.snieskiene@bs.vdu.lt; a.stankeviciene@bs.vdu.lt 

The aim was to investigate the effects of volatile fraction of essential oils from Picea 

abies, Eucalyptus globulus, Rosmarinus officinalis and volatile fraction of Abies sibirica oil 

on fungi isolated from apple, leek, carrot, onion. Researches were made in 2008–2009. Tested 

fungi were as follows: Penicillium roqueforti, Aspergillus flavus, Aspergillus flavus var. oryzae, 

Mortierella hyalina var. hyaline, Sclerotinia sclerotiorum, Sporotrichum aurantiacum, Phoma 

exiqua, Clonostachys rosea f. catenulata. All parts of the plant from which fungi were isolated 

were injured by rot. Fungi of the tested species grew on the potato dextrose agar medium in 

different colonies, which grew at different speed. The oils were dripped on the covers of Petri 

dishes. There were three different variations taking the different portions of oil: 0.005, 0.01 

and 0.015 ml. It was calculated the inhibitory activeness (R). Volatile fractions of all tested 

oils inhibited the growth of mycelium of all 8 species fungi. The inhibiting effect depended 

on: 1) the amount of oil, 2) the species of the plant, from which the oil was isolated, 3) the 

species of the tested fungi and 4) the incubation period. 

Key words: essential oil, fungi, inhibitory activeness. 

Introduction. Essential oils – volatile substance, which consists of many components 

and not all of them composition is known so long. Chemically, essential 

oils are the mixture of various substances and their combinations (Йopдaнoв et al., 

1976; Ragažinskienė et al., 2005). These fractions are responsible for the distinctive 

smell of each plant. Related plants (of the same family or genera) produce different 

fractions. Plants of some families are distinguished for especial richness of oils. One 

of these families is labiates (Labiatae Lindl.), also large amount of essential oils is 

isolated by conifers and eucalyptuses. 

Plants produce volatile fractions, which have been used for various purposes for 

almost 4000 years (Hansel et al., 1999). Antimicrobial features and amounts of essential 

oils isolated from the different plants vary; therefore, laboratory researches in vitro are 

constantly carried out. It is important to test the effect of essential oils of many plants 

227

on different microorganisms. The interest in essential oils as antimicrobial material 

is still growing. In Lithuania and also abroad new researches are made, the effect of 

essential oils of various plants on different microorganisms is tested. 

The essential oils of some plants are used for drugs, also as flavouring and for 

food (Holeman et al., 1984; Šarkinas, Šipailienė, 2003; Tуманова, 2005), in perfumery, 

medicine, especially in aromatherapy (Motiejūnaitė, Pečiulytė, 2004; Kühne, Friedrich, 

2007). The effect of essential oils was tested on fungi isolated from indoor environment 

(Motiejūnaitė, Kalėdienė, 2003; Moтеюнайте, Пячюлите, 2004; Mickienė 

et al., 2007). The results suggest that the essential oils inhibit the growth of most of 

the microorganisms and the effect on some fungi is fungicidal. 

Also one of the areas where essential oils can be applied is plant protection. It is 

considered that among other functions essential oils are helpful to protect plants from 

disease agents and pests. Researches of other authors confirm this (Simoni et al., 1993; 

Klimach et al., 1996; Antonov et al., 1997; Bartynska, 1999; Snieškienė et al., 2008). 

Nevertheless, so far the possibilities of essential oils usage in plant protection have 

been less explored compared with other areas. 

One of the most important causes, which interrupts the usage of the essential oils 

for plant protection, is a quick fragmentation of volatile fraction of oils, dispersion in 

the environment. These features are not such an impediment if oils are used in a room 

as fruit and vegetable storage. It is important to preserve the agricultural production 

using as far as possible less chemicals harmful to human health. 

Antimicrobial features characterize not only essential oils but also common oils 

of some plants. One of these oils is oil from Siberian fir (Abies sibirica). 

The aim of work was to define the effect of volatile fraction of essential oils of 

Picea abies, Eucalyptus globulus, Rosmarinus officinalis and volatile fraction of Abies 

sibirica oil on fungi (8 species) isolated from apple, leek, carrot and onion. 

Object, methods and conditions. Research was done at the Lithuanian Institute 

of Horticulture and Kaunas Botanical Garden of Vytautas Magnus University 

during 2008–2009. The following fungi were tested: Penicillium roqueforti Thom 

was isolated from apple; Aspergillus flavus Link. – from leek and onion; A. flavus 

var. oryzae (Ahlb.) Kurzman, M. J. Smiley, Robnett Wicklow (sin. A. oryzae (Ahlb. 

E. Cohn) – from carrot; Mortierella hyalina var. hyalina (Harz) W. Gams (sin. Mortierella 

hygrophila Linnem., M. hyalina (Harr) W. Gams) – from apple; Sclerotinia 

sclerotiorum (Lib.) de Bary – from carrot; Sporotrichum aurantiacum (Bull.) Fr. – 

carrot; Phoma exiqua Sacc – from apple; Clonostachys rosea f. catenulata (J. C. Gilman, 

E. V. Abbott) Schroers (sin. Gliocladium catenulatum J. C. Gilman, E. V. Abbott) 

– from carrot. Fungi were identified according to Klich (2007); Gams, (1971); 

Lugauskas et al. (2002); Studies …, (1981); Samson, Pitt (2000), currant names are 

given according to Index Fungorum (2004). 

Essential oils used in the study were isolated from three plant species: Picea abies 

Karst., Eucalyptus globulus Labill. and Rosmarinus officinalis L. (oils manufacturer 

Sensient Ess. Oils GmgH, Germany). Also there was tested the volatile fraction of 

Siberian fir (Abies sibirica) oil. The oil is extracted from thorns of young sprouts 

(producer “ALMEDA”, Russia). This oil is recommended as antiseptic in medicine 

and perfumery. 

228

Fungi were grown in Petri dishes on the potato dextrose agar medium (Билай, 

1982). The oils were not added into the medium but dripped on the covers of Petri 

dishes. There were three different variations taking the different portions of oil: 0.005, 

0.01 and 0.015 ml. After sowing the fungi and dripping oil, the dishes were sealed using 

the adhesive tape, turned over and put into a thermostat (temperature 26 ± 2 °C). The 

tests on the effect of the oil were started by measuring the diameter of fungi colonies 

after four days of incubation and by comparing them to the control sample (dishes 

with fungi but without oil). Fungi of the tested species produce different colonies, 

which grow at different speed. For instance the average growth rate of mycelium after 

four days of incubation in the control dishes were: Penicillium roqueforti – 8.3 cm, 

Aspergillus flavus – 50.3 cm, Aspergillus flavus var. oryzae – 32.8 cm, Mortierella 

hyalina var. hyaline – 80.3 cm, Sclerotinia sclerotiorum – 85.0 cm, Sporotrichum 

aurantiacum – 35.0 cm, Phoma exiqua – 25.0 cm, Clonostachys rosea f. catenulate – 

21.0 cm. Therefore, to compare the effect of the oils on the growth of various fungi 

mycelium the inhibitory activeness (%) was calculated using the formula (Билай, 

1982): 

R = (Do - D) / Do × 100 

R – inhibitory activeness (%) 

Do – diameter of a control colony (cm), 

D – diameter of a tested colony (cm). 

Results. Volatile fractions of all tested essential oils and Siberian fir oil inhibited 

the growth of mycelium of all eight species of fungi. 

The bigger concentration (amount of the oil) of volatile oil was in the sealed 

dishes the weaker was the radial growth of fungi mycelium. The growth of fungi of 

all species was the least in the dishes with the biggest oil amount used in a research 

(0.015 ml) (Table, Fig. 1, 2). 

Table. Effect of essential oils of Picea abies, Eucalyptus globulus and Rosmarinus 

officinalis L. on fungi isolated from vegetables and apples 

Lentelė. Picea abies, Eucalyptus globulus ir Rosmarinus officinalis eterinių aliejų poveikis 

grybams, išskirtiems iš daržovių ir obuolių 

Amount of Eucalyptus Rosmarinus Picea 

essential oil globulus 

officinalis abies 

Fungi 

Eterinio 

Inhibitory effect after days 

Grybai 

aliejaus kiekis 

Inhibicinis aktyvumas po dienų (%) 

(ml) 4 7 14 4 7 14 4 7 14 

1 2 3 4 5 6 7 8 9 10 11 

Aspergillus 

flavus 

A. flavus var. 

oryzae 

0.005 75.1 c 72.4 d 22.0 81.8 31.8 60.1 12.9 55.5 0 

0.01 89.5 d 74.4 d 33.8 100 100 18.5 18.5 81.8 3.2 

0.015 90.5 d 90.8 e 41.2 100 100 100 80.1 91.2 47.1 

0.005 41.2 a 26.2 a 0 54.9 47.5 0 4.6 3.3 0 

0.01 100 e 83.7 e 3.5 80.8 59.6 13.2 36.0 9.2 0 

0.015 100 e 91.9 e 13.8 100 100 86.1 52.7 25.9 0 

229



1 2 3 4 5 6 7 8 9 10 11 

0.005 66.7 b 43.5 b 51.3 100 100 77.6 60.5 51.1 40.1 

0.01 100 e 100 e 100 100 100 100 100 70.1 47.9 

0.015 100 e 100 e 100 100 100 100 100 85.6 57.8 

Clonostachys 

rosea f. 

catenulata 

Mortierella 

hyalina var. 

hyalina 

Penicillium 

roquefortii 

Phoma 

exiqua 

Sclerotinia 

sclerotiorum 

Sporotrichum 

aurantiacum 

0.005 75.3 c 38.2 b 0 89.4 82.0 37.1 40.8 0 0 

0.01 100 e 61.8 c 0 100 90.8 51.2 58.5 0 0 

0.015 100 e 100 e 39.6 100 95.5 60.6 71.6 0 0 

0.005 47.9 a 58.8 c 2.7 81.8 31.8 60.1 100 55.3 36.2 

0.01 75.8 c 41.2 b 29.3 100 100 100 100 72.9 50.5 

0.015 100 e 100 e 60.1 100 100 100 100 82.4 62.8 

0.005 65.5 b 49.4 b 30.1 70.0 52.0 45.0 40.7 26.9 14.0 

0.01 100 e 93.3 e 61.2 80.0 56.0 40.0 49.1 33.1 24.0 

0.015 100 e 100 e 64 90.0 60.0 32.0 67.3 46.9 30.1 

0.005 83.5 d 39.6 b 0 100 61.8 25.5 0 0 0 

0.01 100 e 91.2 e 21.2 100 100 87.9 25.3 0 0 

0.015 100 e 96.1 e 33.8 100 100 100 35.3 0 0 

0.005 100 e 83.1 e 52.9 100 100 100 27.7 21.2 19.6 

0.01 100 e 100 e 100 100 100 100 44.9 33.9 23.5 

0.015 100 e 100 e 100 100 100 100 53.4 48.3 26.1 

Essential oils are volatile and fissile fractions and their concentration in the 

environment after some time increased, ant the inhibitory effect increased also. The 

effect of all three species of essential oils was the greatest after four days incubation 

(R from 100 %) and later decreased. After seven days M. hyalina var. hyalina and 

S. sclerotiorum and after fourteen days A. flavus var. oryzae (in all test treatments) 

the space of the mycelium in the dishes with spruce (Picea abies) essential oil and 

S. sclerotiorum and A. flavus var. oryzae with Siberian fir oil have matched the mycelium 

area in control dishes. 

Volatile fraction of Rosmarinus officinalis essential oil had the strongest fungistatic 

and fungicidal effect on all of the investigated micromycetes. Especially sensitive 

to these fractions was C. rosea f. catenulata and S. aurantiacum. P. exiqua (R up to 

32–45 %) was the most resistant to the effect of Rosmarinus officinalis essential oil. 

Volatile fraction of Picea abies essential oil had the least fungistatic effect on all 

of the investigated fungi (Table). 

Fungi of different species have different reaction on different essential oils. From 

the investigated 8 fungi species the most resistant for all essential oil were M. hyalina 

var. hyaline, and sensitive for the influence of volatile fractions of all essential oils 

were C. rosea f. catenulate, P. roquefortii and S. aurantiacum. 

Volatile fractions of Siberian fir oil affected investigated fungi not very evenly. 

Mycelium of P. roquefortii responded firmly to the volatile fraction of this oil 

(Fig. 1). 

230

Fig. 1. Effect of essential oils on the growth of Penicillium roquefortii 

mycelium (%) 

1 pav. Eterinių aliejų poveikis Penicillium roquefortii grybienos augimui, % 

Fig. 2. Effect of essential oils on the growth of 

Mortierella hyalina var. hyalina mycelium (%) 

2 pav. Eterinių aliejų poveikis Mortierella hyalina var. hyalina grybienos augimui, % 

231

A. flavus responded a little bit less negatively to the volatile fractions of this oil. 

The impact persisted for quite a long time: after 14 days of incubation R have increased 

from 47.1 % (0.005 ml) to 67.6 % (0.015 ml). The oil of Siberian fir had almost no 

effect on the growth of M. hyalina var hyalina (Fig. 2). Fungus of this species was 

also resistant to the influence of essential oils of another conifer Picea abies. It can be 

hypothesized that M. hyalina var. hyalina is resistant to the volatile fractions of conifer 

essential oils. Investigated oils of other plants (Rosmarinus officinalis and Eucalyptus 

globulus) strongly inhibited the growth of this fungus (Fig. 2). 

The inhibiting effect volatile fractions of all the tested essential oils and Siberian 

fir oil depended on: 1) the amount of oil, 2) the species of the plant, from which the 

oil was isolated, 3) the species of the tested fungi and 4) the incubation period. 

Discussion. Many authors from different countries have described in their works 

the influence of essential oils on the microorganisms. Essential oils are complex substances; 

therefore, various effect manners of essential oils are tested (Билай, 1982; 

Antonov et al., 1997; Moтеюнайте, Пячюлите, 2004). We have tested the effect of 

essential oils and Siberian fir volatile fraction. Though volatile fractions stay in the 

environment not for a long time, yet the efforts are being made to use them in practice 

in Lithuania – for prophylaxis against bird diseases (Mickienė et al., 2007). Further 

researches are needed to discover the proper essential oils to see their usage possibility 

in vegetable protection. According to our current and previous works (Snieskienė 

et al., 2003; Snieškienė et al., 2008), there can be selected effective essential oils 

against particular fungi species isolated from putrescent vegetables and fruits. 

The volatile fraction of Rosmarinus officinalis and Eucalyptus globulus most 

strongly inhibited the mycelium growth of all fungi (Mortierella hyalina var. hyalinea, 

Penicillium roquefortii and Phoma exiqua) isolated from apples. Fungi (Aspergillus 

flavus var. oryzae, Clonostachys rosea f. catenulate, Sclerotinia sclerotiorum and 

Sporotrichum aurantiacum) isolated from carrots are also strongly affected by the 

same essential oils. Aspergillus flavus, isolated from leeks and onions, is affected by 

all tested essential oils of a higher concentration (0.01 ml and 0.015 ml). 

Conclusions. 1. Fungi of different species have different reaction on different 

oils. 

2. Essential oils of all investigated plants (Rosmarinus officinalis, Eucalyptus 

globulus and Picea abies) and cedar oil had the fungistatic effect. The strongest effect 

(fungistaticaly and fungicidicaly) on fungi was exerted by the volatile fraction 

of Rosmarinus officinalis essential oils. The mentioned oil most strongly affected 

Sporotrichum aurantiacum Penicillium roquefortii and Clonostachys rosea f. catenulate. 

3. The strength of the effect of oils depended on the amount of volatile fraction in 

fungi area: the greater the concentration the stronger was the inhibitory effect. 

Gauta 2009 06 30 


232

References 

1. Antonov A., Stewart A., Walter M. 1997. Inhibition of conidium germination 

and mycelial growth of Botrytis cinerea by natural products. Practical 50th New 

Zealand Plant Protection Conference, 159–164. 

2. Bartynska M. 1999. Effectiveness of essential oils in the control of fungi isolated 

from orchid plants. Bulletin of the Polish Academy of Sciences. Biological 

Sciences, 47(2–4): 123–127. 

3. Gams W. 1971. Hyphomycetes. WEB Gustav Fischer Verlg, Jena. 

4. Hansel R., Sticher O., Steinegger E. 1999. Pharmacognosie-Phytopharmazie, 

Berlin. 

5. Holeman M., Berrada M., Bellakhdar J., Ilidrissi A., Pinel R. 1984. Qualitative 

and quantitative analysis of the essential oil of Salvia officinalis L. from Morocco. 

Boletim da Sociedade Broteriana, 57: 61–67. 

6. Index Fungorum. CABI Bioscience Databases. 2004. www.indexfungorum.org 

7. Klich M. A. 2007. Identification of common Aspergillus species. Centraalbureau 

voor Chimmelcultures, Ultrecht. 

8. Klimach A., Gora J., Wieczorek W. 1996. Wplyw olejkow eterycznych na organiczanie 

wystepowania niektorych chorob grzybowych i bakteryjnych roslin. 

Pestycydy. 1: 45–54. 

9. Kűhne S., Friedrich B. 2007. Pflanzenöle. http://www.bba.de/oekoland/oeko3/ 

pfl-oele.htm 

10. Lugauskas A., Paškevičius A., Repečkienė J. 2002. Patogeniški ir toksiški mikroorganizmai 

žmogaus aplinkoje. Alderija, Vilnius. 

11. Mickienė R., Šiugždaitė J., Bakutis B. 2007. Eterinių aliejų poveikis mikromicetams, 

išskirtiems iš paukštynų oro. Veterinarija ir zootechnika, 40(62): 49–54. 

12. Motiejūnaitė O., Kalėdienė L. 2003. Antimicrobial activity of Lamiaceae plant 

essential oils on Aspergillus niger growth. Bulletin of the Polish Academy of 

Sciences. Biological Sciences, 51(3): 59–64. 

13. Motiejūnaitė O., Pečiulytė D. 2004. Pinus sylvestris L. fungicidai – patalpų oro 

kokybei gerinti. Medicina, 40(8): 787–794. 

14. Ragažinskienė O., Rimkienė S., Sasnauskas V. 2005. Vaistinių augalų enciklopedija. 

Lututė, Kaunas. 

15. Samson R. A., Pitt J. I. (ed.). 2000. Integration of modern Taxonomic methods 

for Penicillium and Aspergillus classification. Centraalbureau voor 

Chimmelcultures, Baarn. 

16. Simoni M., Reuveni R., Ravid U. 1993. Growth inhibition of plant pathogenic 

fungi by essential oils. Hassadeh, 74(3): 306–308. 

17. Snieškienė V., Stankevičienė A., Juronis V. 2003. Growth of micromycetes fungi 

in the presence of essential oils. Bulletin of the Polish Academy of Sciences. 

Biological Sciences, 51(3): 281–285. 

18. Snieškienė V., Stankevičienė A, Varkulevičienė J. 2008. The effect of the essential 

oils on micromycetes isolated from plants. Žemdirbystė, 95(3): 447–452. 

233

19. Studies Mycology 3. 1981. The Phoma and Ascochyta species described by 

Wolenweber and Hochapeel in their study on fruit-rotting. Centraalbureau voor 

Chimmelcultures, Baarn. 

20. Šarkinas A., Šipailienė A. 2003. Maisto saugumo didinimas natūralių medžiagų 

priedais. Veterinarija ir zootechnika, 22(44): 29–32. 

21. Билай В. И. (ред.). 1982. Методы экспериментальной микологии. Hayкoвa 

дyмкa, Киев. 

22. Йopдaнoв Д., Никoлoв П., Бoйчикoв A. 1976. Фитoтepaпия. Meдицинa и 

физкyльтypa, Coфия. 

23. Moтеюнайте О., Пячюлите Д. 2004. Фунгицидные свойства можжевельников 

(Juniperus). Успехи медицинской микологии. 3: 65–67. 

24. Tуманова Е. 2005. Состав эфирных масел растений сем. Яснотковые в 

среднетаежной подзоне республики Коми.




Investigation of pesticides effect on pollination of 

bumblebees in greenhouse tomatoes 

Elena Survilienė, Laimutis Raudonis, Julė Jankauskienė 


Lithuania, e-mail e.surviliene@lsdi.lt 

The study was conducted in three 400–800 m - ² type greenhouses: Multi Rovero 640 TR, 

Rovero 961 and Multispan 9.60 SR of the Lithuanian Institute of Horticulture in 2007. Trial 

data shows that the efficiency of tomato pollination by bumblebees during cultivation season 

ranged from 83.3 to 87.2 %. The pollination is effective, if there are found 60 % of flowers 

pollinated by bumblebees. An efficiency of pollination depends on used pesticides and number 

of hives. Insecticides Aztek 140 EC (triazamat) a concentration of 1.0 ml l -1 , NeemAzal-T/S 

(azadirachtin A) of 5.0 ml l -1 and fungicides Previcur 607 SL (propamocarb hydrochloride) of 

1.5 l ha -1 had not negative affect to bumblebee activity and efficiency of pollination. The effect 

of pollination and activity of bumblebees was reduced due to the use of Euparen M 50 WG 

(tolylfluanid) at the rate of 1.5 mg l -1 . 

Key words: Bombus terrestris, fungicide, insecticide, Lycopersicon esculentum, pollination. 

Introduction. To maximize fruit set in tomatoes and other crops, plant growth 

bioregulator (Pak, Kim, 1999), plant or truss vibration, honeybees and bumblebees 

(Banda, Paxton, 1991; Ilbi, Boztok, 1994; Paydas et al., 2000; Dag, Kammer, 2001) 

are frequently used. In the last years the use of Bombus terrestris (L.) colonies is 

more and more widespread in the pollination of protected crops. The results indicated 

that bumblebees could be used successfully in greenhouses for tomato plant pollination. 

Bumblebee-pollinated tomatoes gave higher yield, higher number of seeds, 

better weight-size correlation, higher specific gravity and higher fruit firmness than 

other pollinating agents; plant growth bioregulator and plant vibration (Banda, Paxton, 

1991; Ravestijn van, Steen van der, 1991; Morandin et al., 2001; Al-Attal et al., 

2003). 

During their foraging behaviour, bumblebees are exposed to the risk of poisoning 

due to pesticide treatments. Therefore, it is necessary to assess the possible dangerousness 

of pesticides. Such a research was begun in some European countries and 

permitted to prepare experimental protocols and to evaluate the toxicity of some of 

the most commonly used active compounds; the results have been recently reviewed 

and discussed by Thompson and Hunt (1999), Thompson (2001) and van der Steen 

235

(2001). The toxicity towards Bombus terrestris (L.) of various insecticides and acaricides 

widely used on protected crops was tested at the laboratory by oral, topic contact, 

and indirect contact trials (Marletto et al., 2003). 

Knowledge of the effects of pesticides on biological control agents is required 

for the successful implementation of integrated pest management (IPM) programs in 

greenhouse production systems. 

The aim of this study was to investigate the influence of pesticides widely used 

on protected crops on effectiveness of bumblebee (Bombus terrestris) pollination in 

tomato production. 

Object methods and conditions. The study was conducted in three 400–800 m - ² 

type greenhouses: Multi Rovero 640 TR, Rovero 961 and Multispan 9.60 SR of the 

Lithuanian Institute of Horticulture in 2007. Seedlings of tomato varieties ‘Cunero’ 

F 1 

were transplanted in rockwool into Multi Rovero 640 TR and Rovero 961 on 

27 of March, in peat bag into Multispan 9.60 SR greenhouse on 19 of March at the 

density of 2.5 plants m -2 . Tomato plants were grown up to the 15 cluster, and then the 

apical growing points were removed to stop plant growth. 

The bumblebee colonies used in the experiment were got from Biobest Belgium 

N. V. The hives were placed by following requirements: the first ones were introduced 

on 24 of April, when 10 % of flowers were opened, and additionally – on 8 of June 

and 30 of July. 

During research time tomato plants were treated with insecticides and fungicides. 

A concentration of 1.0 ml l -1 of Aztek 140 EC (active ingredient triazamat) was sprayed 

only in Multispan 9.60 SR greenhouse on 28 of May; 5.0 ml l -1 of NeemAzal-T/S (a. i. 

azadirachtin A) was sprayed in all greenhouse on 11 of June and 16 of July; 1.5 l ha -1 

Previcur 607 SL (a. i. propamocarb hydrochloride) was used by drip irrigation system 

on 9 of August and 1.5 mg l -1 of Euparen M 50 WG (a. i. tolylfluanid) was sprayed 

on 20 of August. 

The control and effectiveness of pollination were estimated on 10 closed tomato 

flowers in 10 places every week by monitoring bite marks on flowers according to 1–5 

points scale: 1 – black marks, damage on the pistil possible; 2 – brown marks; very 

well pollinated; 3 – flowers are slightly marked; well pollinated; 4 – not all flowers 

are marked; poorly pollinated; 5 – no bite marks; no pollination. The first observations 

were started on 30 of April, the last – on 23 of August. 

Data statistically were analyzed using Fisher LSD 0.5 

test. 

Results. I n v e s t i g a t i o n o f t o m a t o f l o w e r p o l l i n a t i o n. According 

to the efficiency of pollination, not only activity of bumblebees in greenhouses 

was analysed, but also introduction of new hives. Trial data shows that during all activity 

of bumblebees the effectiveness of pollination of tomato flowers was high. Efficacy 

of pollination in Multi Rovero 640 TR greenhouse on average reached 84.7 % 

(Fig. 1), Rovero 961 – 87.2 % (Fig. 2) and Multispan 9.60 SR – 83.3 % (Fig. 3). Air 

temperature was not higher than 25 °C, and relative humidity ranged from 78–85 % 

during all the investigation period. 

Efficiency of pollination of first hive during six weeks in Multi Rovero 640 TR 

reached on average 81.8 %, the quality of pollination corresponded to 2–3 points, which 

236

mean very well and well pollination. The lowest activity of bumblebees was during 

sixth week, the efficiency of pollination was only 56.7 %, the quality of pollination 

corresponded to 4th point, which means poorly pollination (Fig. 1). After introduction 

of additional hive on 8 of June, efficiency of pollination increased by 31.3 %. Activity 

of bumblebees of this hive during seven weeks reached on average 79.9 %. The efficiency 

of third hive activity was observed from 31 of July to 23 of August. During 

three weeks the efficiency of pollination was high and reached on average 92.5 %. 

Fig. 1. Efficiency of pollination using bumblebees in 

Multi Rovero 640 TR greenhouse, 2007. LSD 0.5 

– 7.9 

(treatment with NeemAzal-T/S on 11 of June, on 16 of July and 

with Previcur 607 SL – on 9 of August) 

1 pav. Pomidorų apdulkinimo efektyvumas naudojant kamanes Multi Rovero 640 TR šiltnamyje, 

2007 m. R 0,5 

– 7,9 (apdorota Nimazaliu T/S 10 g/l k.e. birželio 11 d., liepos 16 d. ir 

Previkuru 607 g/l v. t. rugpjūčio 9 d.) 

Fig. 2. Efficiency of pollination using bumblebees in Rovero 961 greenhouse, 

2007. LSD 0.5 

– 8.61 (treatment with NeemAzal-T/S on 11 of June and 

Previcur 607 SL on 9 of August) 

2 pav. Pomidorų apdulkinimo efektyvumas naudojant kamanes Rovero 961 šiltnamyje, 

2007 m. R 0,5 

– 8,61 (apdorota Nimazaliu T/S 10 g/l k.e. birželio 11 d. ir 

Previkuru 607 g/l v. t. rugpjūčio 9 d.) 

237

Efficiency of bumblebees from the first introduced hive during six weeks in 

Rovero 961 greenhouse reached on average 86.1 % (2–3 pollination quality points). 

The lowest pollination was during sixth week of introduction, the efficiency was 

77.5 % or 4 points of pollination quality (Fig. 2). Second additional introduction of 

hive increased efficiency of pollination by 19.17 %. Efficiency of pollination during 

seven weeks totally reached on average 83.4 % and only later decreased to 46.3 %. 

The third additional introduction of hive increased efficiency during three weeks from 

88 to 100 %. 

Analogical situation was in Multispan 9.60 SR greenhouse. After introduction 

of the first hive of bumblebees, pollination efficiency during six weeks reached on 

average 85.8 % (2–3 pollination quality points) (Fig. 3). The activity of bumblebees 

decreased in the end of sixth week and the efficiency of pollination reached 76.7 %. 

After additional second and third introduction of hives, the efficiency of bumblebee 

activity on average was 78.4 % and 85.7 %, respectively. 

Fig. 3. Efficiency of pollination using bumblebees in Multispan 9.60 SR greenhouse, 

2007. LSD 0.5 

– 15.62 (treatment with Aztek 140 EC on 28 of May, 

NeemAzal-T/S on 11 of June, Previcur 607 SL on 9 of August and 

Euparen M 50 WG on 20 of August) 

3 pav. Pomidorų apdulkinimo efektyvumas naudojant kamanes Multispan 9.60 SR šiltnamyje, 

2007 m. R 0,5 

– 15,62 (apdorota Acteku 140 g/l v. e. gegužės 28 d., 

Nimazaliu T/S 10 g/l k. e. birželio 11 d., Previkuru 607 g/l v. t. rugpjūčio 9 d. ir 

Euparenu M 50 % v. t. g. rugpjūčio 20 d.) 

S i d e - e f f e c t o f p e s t i c i d e s o n b u m b l e b e e s. Aficide Aztec 140 EC 

applied only in hot spots at the rate of 1.0 ml l -1 solution had very small side-effect 

on activity of bumblebees. Efficiency of pollination was reduced only by 5 %. This 

reduction can be explained as natural (Fig. 3). The activity of bumblebees was reduced 

to 56.7 % in Multi Rovero 640 TR, and to 77.5 in Rovero 961 greenhouses where 

Aztec 140 EC was not used (Fig. 1, 2). Insecticide NeemAzal-T/S used for the first 

time did not has any side-effect on bumblebees or pollination of flowers. Efficacy of 

238

pollination of flowers after a day and later was about 90 %. The second NeemAzal-T/S 

application superposed with the end of bumblebee life in hive; therefore, the activity 

of bumblebees reached only 42 % on 23 of July (Fig. 1). Efficiency of pollination of 

tomato flowers in Rovero 961 and Multispan 9.60 SR greenhouses shows that activity 

of bumblebees depends on life time of hives, not on the NeemAzal-T/S used. Though 

NeemAzal-T/S was not used additionally the efficacy of pollination on 23 of July 

reached only 45–46 % (Fig. 2, 3). 

Previcur 607 SL used by drip irrigation system on 9 of August in all types of 

greenhouses did not have any negative impact on activity of bumblebees. Efficiency 

of pollination reached about 90 % (Fig. 1–3). 

Euparen M 50 WG (1.5 mg l -1 ) used on 20 of August on the average reduced efficiency 

of tomato pollination in Multispan 9.60 SR greenhouse to 53 %; meanwhile, 

where Euparen M 50 WG was not used in other greenhouses efficiency was 96.7–100 % 

(Fig. 1, 2). 

Discussion. Pollination of flowers and activity of bumblebees depends on life 

time of hive. It was noticed the reduction of efficiency of pollination by bumblebees 

in greenhouse after 5–6 weeks. Efficiency of pollination on 4 of June was 56.7 % 

in Multi Rovero 640 TR, 77.5 % in Rovero 961 and 76.7 % in Multispan 9.60 SR 

greenhouses. Efficiency of pollination on 20 of July (before introduction of additional 

hives) was 41.7 % in Multi Rovero 640 TR, 46.3 % in Rovero 961 and 45 % in 

Multispan 9.60 SR greenhouses (Fig. 1–3). Additional hives should be introduced in 

greenhouse when efficiency of pollination is highest (Banda, Paxton, 1991; van Ravestijn, 

van der Steen, 1991; Morandin et al., 2001). Continuously equal pollination 

of flowers is available only when additional hives are introduced at the beginning of 

reduction of activity of pollination. In our case hives had to be changed 1–3 weeks 

earlier to avoid sudden reduction of pollination. 

Beneficial fauna and other products are used in greenhouses to control pests for 

high and good quality of production. It is very important to make trials to know transaction 

between beneficial organisms and pesticides controlling the pests according to 

the integrated pest management practices. It is important to select compatible fungicides 

and insecticides with bumblebees, which are used for pollination, because a lot of 

chemicals are toxic or could have repellent properties for adults, hatch of bumblebees 

(Hassan, 1996; Steen van der, 2001). 

Insecticides and acaricides both from chemical and biological origins were tested 

for their toxicity to bumblebees and beneficial organisms. All microbial compounds 

were very safe for all the tested species. Azadirachtin, indoxacarb, pymetrozine and 

spinosad were almost completely harmless, although spinosad was moderately toxic 

to adult Encarsia formosa, but just like the effect on bumblebees, the persistence in 

practice was very short. The neonicotinoids, imidacloprid and thiamethoxam, were 

very toxic. Acetamiprid and thiacloprid on the other hand were safer. Fipronil was 

very toxic for all organisms (Sterk et al., 2003). 

As described in “Side effect manual” (Sterk, Put, 2004) active ingredient of 

propamokarb hydrochloride does not have any negative action for bumblebees. 

Azadirachtin does not have side-effect on bumblebees and can be used for integrated 

239

pest management in vegetables. Active ingredient of triazamat when was sprayed or 

plants were watered with it had no negative impact on bumblebees as well. The same 

results were obtained in our study. Other authors stated that azoxystrobin, chlorotalonil, 

copper oxychloride, sulphur, mancoceb, tolylfluanid, iprodione, metalaxyl, dimetomorf, 

penconazole do not have adverse effect on beneficial fauna and could be safely used 

in integrated pest management (Sterk et al., 2003, Sterk, Put, 2004). 

Conclusions. Efficiency of bumblebees during all their activity period was high 

and ranged from 83.3 to 87.2 %. 

Efficiency of bumblebees for pollination of tomato flowers depends on additional 

hives and pesticides used. 

Insecticides NeemAzal-T/S (azadirachtin A) 5.0 ml l -1 , Aztec 140 EC (triazamat) 

at the rate of 1.0 ml l -1 and fungicide Previcur 607 SL (propamocarb hydrochloride) at 

the rate of 1.5 l ha -1 had no negative effect on activity of bumblebees and pollination 

of tomato flowers. 

Spraying tomato with 1.5 mg l -1 solution of fungicide Euparen M 50 WG (tolylfluanid) 

reduced activity of bumblebees and efficiency of pollination. 

Acknowledgement. The study was supported by Lithuanian State Science and 

Studies Foundation. 

Gauta 2009 06 30 


References 

1. Al-Attal Y. Z., Kasrawi M. A., Nazer I. K. 2003. Influence of pollination technique 

on greenhouse tomato production. Agricultural and Marine Sciences, 8(1): 

21–26. 

2. Banda H. J., Paxton R. J. 1991. Pollination of greenhouse tomatoes by bees. 

Acta Horticulturae, 288: 194–198. 

3. Dag A., Kammer Y. 2001. Comparison between the effectiveness of honey bee 

(Apis mellifera) and bumble bee (Bombus terrestris) as pollinators of greenhouse 

sweet pepper (Capsicum annuum). American Bee Journal, 141(6): 447–448. 

4. Ilbi H., Boztok K. 1994. The effects of different truss-vibration durations on 

pollination and fruit set of greenhouse grown tomatoes. Acta Horticulturae, 366: 

73–78. 

5. Marletto F., Paletta A., Manino A. 2003. Laboratory assessment of pesticide toxicity 

to bumblebees. Bulletin of Insectology, 56(1): 155–158. 

6. Morandin L. A., Laverty T. M., Kevan P. G. 2001. Bumble bee (Hymenoptera: 

Apidae) activity and pollination levels in commercial tomato greenhouses. 

Journal of Economic Entomology, 94(2): 462–467. 

240

7. Pak H., Kim D. 1999. Effect of 4–chlorophenoxyacetic acid on fruit set and 

nutrient accumulation in Cucurbita moschata (Duch.) Poir. Acta Horticulturae, 

483: 381–386. http://www.actahort.org/books/483/483_44.htm 

8. Paydas S., Eti S., Kaftanglu O., Yasa E., Derin K. 2000. ISHI Acta Horticulturae 

(On line). http://www.ishs.org/pub/483/513.html 

9. Hassan S. A. 1996. Activities of the IOBC/WPRS Working group “Pesticides 

and Beneficial Organisms”. 178. 

10. Steen J. J. M. van der. 2001. Review of the methods to determine the hazard and 

toxicity of pesticides to bumblebees. Apidologie, 32: 399–406. 

11. Sterk G., Jans K., Put K., Wulandari O. V., Uyttebroek M. 2003. Toxicity of 

chemical and biological plant protection products to beneficial arthropods. In: 

L. Roche, M. Edin, V. Mathieu, F. Laurens (eds.), Colloque international tomate 

sous abri, protection intégrée – agriculture biologique. Centre Technique 

Interprofessionnel des Fruits et Légumes Publisher. 

12. Sterk G., Put K. 2004. Side effect manual. Werantwoordelijke. 29. 

13. Thompson H. M. Hunt L. V. 1999. Extrapolating from honeybees to bumblebees 

in pesticide risk assessment. Ecotoxicology, 8: 147–166. 

14. Thompson H. M. 2001. Assessing the exposure and toxicity of pesticides to 

bumblebees (Bombus sp.). Apidologie, 32: 305–321. 

15. Ravestijn W. van, Steen J. van der. 1991. Use of bumblebees for the pollination 

of glasshouse tomatoes. Acta Horticulturae, 288: 204–212. 


Pesticidų poveikio kamanių entomofilijai šiltnamio pomidoruose tyrimas 

E. Survilienė, L. Raudonis, J. Jankauskienė 

Santrauka 

Tyrimai atlikti Lietuvos sodininkystės ir daržininkystės instituto eksperimentinės 

bazės Multi Rovero 640 TR, Rovero 961 ir Multispan 9.60 SR šiltnamiuose 2007 m. 

Rezultatai rodo, kad per visą kamanių veiklos laiką jos veiksmingai apdulkino pomidorų 

žiedus – 83,3–87,2 %. Kamanių darbo efektyvumui ir apdulkinimo kokybei turi įtakos papildomų 

avilių pastatymas bei šiltnamiuose naudojami pesticidai. Panaudoti insekticidai 

Nimazalis T/S 10 g/l k. e. (azadirachtinas A), išpurkštas 0,5 % normos, Actekas 140 g/l v. e. 

(triazamatas) – 0,1 % normos ir Previkuras 607 g/l v. t. (propamokarbo hidrochloridas) – 

1,5 l/ha normos neturėjo neigiamo poveikio kamanių darbingumui ir pomidorų žiedų apdulkinimui. 

Išpurškus pomidorus 0,15 % normos fungicidu Euparenas M 50 % v. t. g. (tolylfluanidas), 

pastebėtas kamanių veiklos susilpnėjimas ir apdulkinimo efektyvumo sumažėjimas. 

Reikšminiai žodžiai: apdulkinimas, Bombus terrestris, fungicidai, insekcitidai, 

Lycopersicon esculentum. 

241




Effectiveness of Olejan 85 EC against 

chrysanthemum and willow rust 

Adam T. Wojdyła 

Research Institute of Pomology and Floriculture in Skierniewice, 

ul. Pomologiczna 18, 96-100 Skierniewice, Poland, e-mail awojdyla@insad.pl 

For the protection of chrysanthemum against Puccinia horiana, Olejan 85 EC (85 % 

rapeseed oil) was used at concentrations of 0.5 and 2 % and sprayed 4 times every 7 days. 

After four treatments Olejan 85 EC was found to have inhibited the development of Puccinia 

horiana from 1.4 to 3.6 times depending on the concentration and had caused sporadic, up to 

almost 37 % browning and decomposition of telia. 

Protecting willow against Melampsora epitea, Olejan 85 EC was used at concentrations 

of 0.5 and 2 % and sprayed 2 times every 7 days. After two treatments Olejan 85 EC was found 

to have inhibited the development of M. epitea from 2.6 to 13.7 times and had caused from 

10 to 62 % browning and decomposition of uredinia. Olejan 85 EC was significantly more 

effective reducing leaf infection than Saprol 190 EC. Taking into consideration the percentage 

of dried-up uredinia per leaf, Olejan 85 EC had also shown significantly higher efficacy in 

comparison with fungicide Saprol 190 EC. At neither of the concentrations used Olejan 85 

EC was phytotoxic to the treated plants. 

Key words: control, Melampsora epitea, Olejan 85 EC, Puccinia horiana, rust. 

Introduction. Olejan 85 EC (85 % rapeseed oil) has been recommended as an 

adjuvant intended for use in combination with working solutions of some plant protection 

products. Atpolan 80 EC and Olejan 85 EC at a concentration of 0.3 % used 

as additions to working solutions make it possible to reduce the dose of the emulsion 

fungicides recommended for controlling powdery mildew and black spot on rose as 

much as 30–50 % (Orlikowski, Wojdyła, 1995; Wojdyła, 1998; Wojdyła, 1999; Zdonek 

et al., 1986). Current literature data indicate high efficacy of the oils used to control 

many foliar pathogens responsible for plant diseases (Dell et al., 1998; Ko et al., 

2003; Picton, Humer 2003). The author’s own studies carried out during many years 

have also shown high efficacy of mineral and vegetable oils used at concentrations 

of 0.25–4 % controlling foliar pathogens. The oils employed in the protection of rose 

bushes against powdery mildew (Sphaerotheca pannosa var. rosae) were from 90 to 

100 % effective inhibiting the development of disease symptoms (Wojdyła, 2002). In 

the protection of roses against black spot (Diplocarpon rosae) their efficacy ranged 

from 40 to 60 %. When used to control willow rust (Melampsora epitea) (Wojdyła 

243

and Jankiewicz 2004) and Puccinia pelargoni-zonalis on geraniums (Wojdyła, 2005) 

they significantly hampered the development of these pathogens. In the protection of 

roses against grey mould (Botrytis cinerea) their efficacy limiting the extent of necrosis 

on rose petals was in the range of 24–78 % depending on the oil used (Wojdyła, 

2003). 

The aim of the experiments was to evaluate the product Olejan 85 EC in terms 

of its efficacy controlling chrysanthemum rust (Puccinia horiana) and willow rust 

(Melampsora epitea). 

Object, methods and conditions. Control of Puccinia horiana. 

Seedlings of chrysanthemum cv. ‘Fiji Yellow’, susceptible to Puccinia horiana, were 

planted in 1 dm 3 pots filled with peat substrate. The plants were placed in a greenhouse 

on windowsills covered with capillary mats. A chrysanthemum with symptoms 

of rust sporulation was placed among the healthy, newly planted plants. In order to 

ensure air humidity above 90 % favourable to that pathogen’s development the sills 

were covered with a thin-foil tunnel. After the first rust spots had been found on the 

leaves, but before the formation of telia, spraying of the plants began. The chrysanthemum 

plants were sprayed 4 times at 7-day intervals. Before application of the 

controlling agents and then after spray treatments, the percentage of infected leaves 

and the average number and percentage of dried-up telia were determined. 

Control of Melampsora epitea. Willow tree cv. ‘Iwa’ growing on a 

loamy soil in open field, after the first signs of sporulation (clusters of uredinia) of rust 

(M. epitea) had been found on the underside of their leaves, was sprayed twice every 

7 days with Olejan 85 EC at a concentration of 1 % solution. Prior to the experiment 

and after two spray treatments, the average percentage of diseased leaves, average 

number of uredinia formed per leaf, and the percentage of those uredinia that had 

turned brown and decomposed were determined. 

In all the experiments undertaken, plants were sprayed in the morning using 1 dm 3 

of working solution per 10 m 2 of surface area. Both the upper surface and the underside 

of the leaf blade were thoroughly covered. Saprol 190 EC (190 g · L -1 triforine) was 

the standard fungicide used. 

The experiments were set up in a random block design in 4 replications, with 5 

plants (chrysanthemum) or 25 leaves (willow) per replication. 

Results. Control of Puccinia horiana. In the first experiment, when 

the protection programme was finished, it was found that the average percentage of 

infected leaves protected with Olejan 85 EC was similar to that of the control plants 

(Table 1). However, the number of telia on the leaves sprayed with the product depending 

on its concentration was found to be from 2 to 3.6 times lower; from 1 to 

about 45 % of the telia had turned brown and were decomposing. 

In the second experiment, when spray treatments were finished, the average percentage 

of infected leaves of chrysanthemum plants protected with Olejan 85 EC was 

similar, and in the concentrations of 0.5 % and 25 % even significantly higher compared 

with the control chrysanthemums (Table 2). On the leaves of the plants sprayed with 

the product, the number of telia forming per leaf was from 1.4 to 2.1 times lower in 

comparison with the control plants. In the case of chrysanthemum plants sprayed with 

244

Olejan 85 EC at a concentration of 0.5 %, the number of telia per leaf was similar to 

that on the control chrysanthemums. On the plants protected with the product, from 

19 to almost 37 % of telia were found dried-up. 

Table 1. Effectiveness of Olejan 85 EC controlling Puccinia horiana on chrysanthemum 

cv. ‘Fiji Yellow’. Beginning of experiment – 2003.08.01 

1 lentelė. Olejan 85 EC efektyvumas kontroliuojant Puccinia horiana chrizantemose ‘Fiji 

Yellow’. Bandymo pradžia – 2003.08.01 

Treatment 

Variantas 

Control 

Kontrolė 

Saprol 190 EC 

Olejan 85 EC 

Olejan 85 EC 

Olejan 85 EC 

Olejan 85 EC 

Olejan 85 EC 

Concentration 

Koncentracija 

(%) 

- 

0.15 

0.5 

0.75 

1.0 

1.5 

2.0 

Average percentage 

of diseased leaves 

Vidutinis užkrėstų 

lapų skaičius 

62.3 b 

26.5 a 

51.1 b 

55.8 b 

54.0 b 

54.2 b 

59.1 b 

Average number of 

pustules per leaf 

Vidutinis pustulių ant 


11.6 c 

0.7 a 

4.2 ab 

5.9 b 

3.4 ab 

3.2 ab 

3.3 ab 

Average percent of 

dried pustules 

Vidutinis sudžiūvusių 

pustulių procentas 

0.2 a 

0 a 

0.7 a 

4.3 ab 

12.5 b 

43.5 c 

41.8 c 

Note: Mean values marked with the same letter do not differ at the significance level p = 0.05 

according to the Duncan’s test 

Pastaba: Vidutinės reikšmės, pažymėtos ta pačia raide, pagal Dunkano kriterijų nesiskiria, kai 

p = 0,05 

Table 2. Effectiveness of Olejan 85 EC controlling Puccinia horiana on chrysanthemum 

cv. ‘Fiji Yellow’. Beginning of experiment – 2003.08.26 

2 lentelė. Olejan 85 EC efektyvumas kontroliuojant Puccinia horiana chrizantemose ‘Fiji 

Yellow’. Bandymo pradžia – 2003.08.26 

Treatment 

Variantas 

Control 

Kontrolė 

Saprol 190 EC 

Olejan 85 EC 

Olejan 85 EC 

Olejan 85 EC 

Olejan 85 EC 

Olejan 85 EC 

Note: See Table 1 

Pastaba: žr. 1 lentelę 

Concentration 

Koncentracija 

(%) 

- 

0.15 

0.5 

0.75 

1.0 

1.5 

2.0 

Average percentage 

of diseased leaves 

Vidutinis užkrėstų 


65.9 bc 

61.7 a 

72.0 d 

65.2 b 

67.4 bc 

69.0 c 

77.7 e 


pustules per leaf 

Vidutinis pustulių ant 


24.2 d 

18.3 c 

24.7 d 

17.8 c 

13.4 b 

12.3 a 

11.7 a 

Average percent of 

dried pustules 

Vidutinis sudžiūvusių 

pustulių procentas 

0 a 

7.3 b 

25.5 d 

19.3 c 

28.9 e 

29.1 e 

36.6 f 

245

Control of Melampsora epitea. In the first experiment, when protection 

programme was finished almost 92 % of leaves on the control willow were found 

to be infected (Fig. 1). On the other hand, on the plants protected with the product 

being evaluated depending on its concentration only about 44–51 % of leaves were 

found to be diseased. Olejan 85 EC showed similar effectiveness controlling the degree 

of infection on willow leaves in comparison with the product Saprol 190 EC. After 

two spray treatments of willow the plants protected with Olejan 85 EC were found 

to contain from 4.4 to 13.7 times fewer uredinia forming per leaf, almost 10–26 % of 

which were dried-up and decomposing. 

Fig. 1. Effectiveness of Olejan 85 EC controlling Melampsora epitea on willow. 

Beginning of experiment – 2003.08.21 

1 pav. Olejan 85 EC efektyvumas kontroliuojant Melampsora epitea gluosniuose. 

Bandymo pradžia – 2003.08.21 

In the second experiment, when spray treatments were finished almost 100 % of 

diseased leaves were found on the control willows (Fig. 2). 

Fig. 2. Effectiveness of Olejan 85 EC controlling Melampsora epitea on willow. 

Beginning of experiment – 2003.09.07 

2 pav. Olejan 85 EC veiksmingumas kontroliuojant Melampsora epitea ant gluosnių. 

Bandymo pradžia – 2003.09.07 

246

By contrast, on the plants protected with the evaluated product depending on its 

concentration there were about 42–74 % of infected leaves. Olejan 85 EC showed 

significantly higher efficacy controlling the extent of leaf infection than Saprol 190 

EC. After two spray treatments of willow, on the plants protected with Olejan 85 EC 

there were from 2.6 to 10 times fewer uredinia forming per leaf, 17–62 % of which 

had dried up and were decomposing. Also, considering the percentage of dried-up 

uredinia per leaf, Olejan 85 EC was significantly more effective in comparison with 

the fungicide Saprol 190 EC. 

Discussion. Earlier studies had also shown high efficacy of Olejan 85 EC controlling 

willow rust (Wojdyła, Jankiewicz, 2004). The evaluated compound depending 

on its concentration caused 2 to 14-fold reduction in the formation of uredinia 

clusters, about 10–16 % of which were brown and decomposing. Similar experiments 

on geranium confirmed high effectiveness of Olejan 85 EC controlling Puccinia 

pelargonii-zonalis (Wojdyła, 2005). The author showed that the product at a 

concentration of 1 % after spraying geranium plants 4 times every 7 days caused an 

almost twofold reduction in the number of uredinia compared with control plants and 

23 % of uredinia were dried-up. 

Conclusions. 1. Olejan 85 EC used for curative spray treatments of chrysanthemum 

was found to significantly inhibit the development of rust. After 4 treatments 

depending on the concentration Olejan 85 EC caused from 1.4 to 3.6-fold reduction 

in the formation of telia of Puccinia horiana and sporadically caused almost 37 % of 

them to turn brown and decompose. 

2. In the protection of willow, after 2 spray treatments Olejan 85 EC caused from 

2.6 to 13.7-fold reduction in the formation of uredinia of Melampsora epitea and caused 

from 10 to 62 % of them to turn brown and decompose. 

3. No evidence of phytotoxicity of Olejan 85 EC towards chrysanthemum and 

willow cultivars in the experiment was found. 

Gauta 2009 06 30 


References 

1. Dell K. J., Gubler W. D., Krueger R., Sanger M., Bettiga L. J. 1998. The efficacy 

of JMS Style Oil on grape powdery mildew and Botrytis cinerea bunch rot and 

effects on fermentation. American Journal of Enology and Viticulture, 49(1): 

11–16. 

2. Ko W. H., Wang S. Y., Hsieh T. F., Ann P. J., 2003. Effects of sunflower oil 

on tomato powdery mildew caused by Oidium neolycopersici. Journal of 


3. Orlikowski L. B., Wojdyła A. 1995. Adiuwanty w ochronie roślin ozdobnych 

przed chorobami. Hasło Ogrodnicze, 5: 39. 

247

4. Picton D. D., Hummer K. E., 2003. Oil application reduces white pine blister 

rust severity in black currants. Small Fruits Review, 2(1): 43–49. 

5. Wojdyła A. 1998. Chemical control of rose diseases: IV. Effectiveness of fungicides 

in the control of powdery mildew in relation to period leaves wetness or 

Atpolan 80 EC addition. J. Fruit Ornam. Plant Res., 6(3–4): 147–156. 

6. Wojdyła A. T. 1999. Możliwość wykorzystania związków olejowych w ochronie 

roślin ozdobnych przed chorobami. VI Konferencja Szkółkarska “Nowe tendencje 

w szkółkarstwie ozdobnym” Skierniewice, 18–19 listopada 1999, 169–174. 

7. Wojdyła A. T. 2000. Influence of some compounds on development of 

Sphaerotheca pannosa var. rosae. J. Plant Protection Res., 40(2): 106–121. 

8. Wojdyła A. T. 2002. Oils activity in the control of rose powdery mildew. Med. 

Fac. Landbouww. Univ. Gent, 67(2): 369–376. 

9. Wojdyła A. 2003. Biological activity of plant and mineral oils in the control of 

Botrytis cinerea on roses. Bulletin of the Polish Academy of Science – Biological 

Sciences 51(2): 153–158. 

10. Wojdyła A. T. 2005. Activity plant and mineral oils in the control of Puccinia 

pelargonii-zonalis. Comm. App. Biol. Sci. Ghent University, 70(3), 193–198. 

11. Wojdyła A. T, Jankiewicz D. 2004. Oils activity against Melampsora epitea on 

willow. Comm. App. Biol. Sci. Ghent University, 69(4): 697–703. 

12. Zdonek Z., Orlikowski L. B., Wojdyła A. 1986. Zastosowanie olejów w ochronie 

róż. Ogrodnictwo, 11: 28–29. 


Olejan 85 Ec veiksmingumas kontroliuojant chrizantemų ir gluosnių rūdis 

A. T. Wojdyła 

Santrauka 

Saugant chrizantemas nuo Puccinia horiana, 4 kartus kas 7 dienas buvo purkšta 0,5 ir 

2 % koncentracijos Olejan 85 EC (85 % rapsų aliejaus). Po keturių purškimų Olejan 85 EC 

sustabdė Puccinia horiana vystymąsi nuo 1,4 iki 3,6 karto priklausomai nuo koncentracijos ir 

sukėlė sporadišką beveik 37 % telia parudavimą ir suirimą. 

Saugant gluosnius nuo Melampsora epitea, 0,5 ir 2 % koncentracijos Olejan 85 EC buvo 

purkšta 2 kartus kas 7 dienos. Po dviejų purškimų Olejan 85 EC sustabdė M. epitea vystymąsi 

nuo 2,6 iki 13,7 karto ir sukėlė 10–62 % uredinia parudavimą ir suirimą. 

Olejan 85 EC buvo daug veiksmingesnis kovojant su lapų infekcija negu Saprol 190 EC. 

Atsižvelgiant į sudžiūvusios uredinia procentą, tenkantį vienam lapui, Olejan 85 EC taip pat 

buvo daug veiksmingesnis palyginus su fungicidu Saprol 190 EC. Jokia Olejan 85 EC koncentracija 

nebuvo fitotoksiška purkštiems augalams. 

Reikšminiai žodžiai: kontrolė, Melampsora epitea, Olejan 85 EC, Puccinia horiana, 

rūdys. 

248




Application of biological insecticide 

Pecilomicine-B for greenhouse pest control 

Alena Yankouskaya 

RUC Institute of Plant Protection, Priluki, Мinsk region, Belarus, 


The estimation of influence of some technological parameters (terms and number of treatments, 

the interval between them) on biological efficiency of bioinsecticide Pecilomicine-B 

on greenhouse whitefly (Trialeurodes vaporariorum Wеst.) and cucumber midge (Bradysia 

brunnipes Mg.), and also its influence on entomophages encarsia (Encarsia formosa Gahan.) 

and phytoseiulus (Phytoseiulus persimilis Ath.) was carried out under greenhouse conditions. 

It was determined that the most expedient method of Pecilomicine-B application is at 

the stage of primary settling of greenhouse crop plantings by phytophage: in case with greenhouse 

whitefly – by the first imago appearance on plant leaves carrying out 2 treatments 

at 7–14 days interval. Later on, Pecilomicine-B (1 % concentration) is applied up to 4 times 

considering greenhouse whitefly population dynamics (in case its stable increase). It allows 

keeping the phytophage population during 1.5–2 months at economically imperceptible level 

without chemical means of plant protection. Against cucumber midge one should apply the 

preparation (4 % concentration up to 2 times at 23–27 days interval) at the beginning of pest 

imago mass flight. In case of combined Pecilomicine-B application under greenhouse conditions 

(at 7–14 days interval) with phytoseiulus and encarsia it does not render the negative 

influence on survival, reproduction, parasitic and predatory activity of entomophages and 

does not decrease the efficiency of their action. Pecilomicine-B application, according to the 

above mentioned technological parameters, allows to constrain the phytophage populations 

at economic-imperceptible level. 

Key words: bioinsecticide efficiency, bioinsecticide influence on entomophages, Bradysia 

brunnipes, Cucumis sativus, Encarsia formosa, Lycopersicon esculentum, Paecilomyces fumosoroseus, 

Pecilomicine-B, Phytoseiulus persimilis, Trialeurodes vaporariorum. 

Introduction. One of the most rational approaches in greenhouse crop protection 

against pests is a combination of different means of action on phytophages with a 

maximum proportion of ecologically safe biological preparations. In this connection 

it is actual to extend the spectrum of biological insecticides recommended for application. 

The analysis of literary data shows that the entomopathogenic fungus Paecilomyces 

fumosoroseus (Wize) Brown et Smith (Deuteromycota: Moniliaceae) is a 

perspective agent for biological control of greenhouse pests (Ижевский et al., 1999; 

Poprawski, Jones, 2000; Wraight et al., 2000; Faria et al., 2001; Lacey et al., 2001; 

249

Nigroho, Ibrahim, 2004; Ахатов et al., 2004; Thungrabeab et al., 2006; Shi, Feng, 

2009). As a result of done screening by insecticidal activity level of P. fumosoroseus 

strains from the collection of the Institute of Plant Protection, there was selected 

P. fumosoroseus 3/1 strain possessing high virulence in relation to the most noxious 

greenhouse phytophages in Belarus: greenhouse whitefly Trialeurodes vaporariorum 

Wеstw. and cucumber midge Bradysia brunnipes Mg. (Прищепа, Янковская, 2008). 

This strain is a basis of mycoinsecticidal preparation Pecilomicine-B (a paste with the 

spore titer 1.8 ·10 10 /g, manufacturer RUC “Novopolotsk Plant of Protein and Vitamin 

Concentrates”) (Прищепа, 2005; Янковская, 2006; Янковская, Прищепа, 2008). 

To develop the optimum technological techniques of preparation application we evaluated 

the influence of different factors (time and number of treatments, the intervals 

between them) on Pecilomicine-B biological efficiency and also the character of interrelation 

with other elements of biological protection and especially with the most 

frequently applied in the greenhouses of Belarus entomophages encarsia (Encarsia 

formosa Gahan.) and phytoseiulus (Phytoseiulus persimilis Ath.). 

Object, methods and conditions. The researches were carried out in greenhouses 

of Minsk region greenhouse farm “DorOrs”, “Zhdanovichy”, “Oziorny” and in 

greenhouses of the Republican Ecological Educational Center on tomato (‘Raissa’ F 1 

, 

‘Barcelona’ F 1 

, ‘Blagovest’ F 1 

), cucumber (TSHA 14-27 F 1 

, ‘Mystica’ F 1 

, ‘Turnir’ F 1 

) 

and soybean crops. The preparation was applied by the method of spraying (preparation 

concentration – 1 %) against greenhouse whitefly and by the method of watering 

in the collar zone of cucumber (from the calculation of 50 ml/plant with preparation 

concentration 4 %) against cucumber midge. Time and number of treatments were 

determined in the course of experiments considering pest number changes. Pest number 

was registered every week and the biological efficiency of preparation action was 

evaluated according to the general methods (Прищепа et al., 2008). The record of 

parasitized whitefly larvae and flying out encarsia imago was done in the lab under 

binocular. 

Results. The efficiency of Pecilomicine-B application against greenhouse whitefly 

was evaluated under greenhouse conditions in 2002–2006. In each separate case 

a scheme of preparation application considering the dynamics of phytosanitary condition 

of greenhouse crop plantings was developed. 

The efficiency of Pecilomicine-B preparation application against greenhouse 

whitefly was evaluated under greenhouse conditions in 2002 in the greenhouse farm 

“DorORS” on tomato crop ‘Raissa’ F 1 

, (small-volume technology of cultivation on 

rock wool). In total for the registration period there were 3 treatments. The control plot 

was without any treatment. The first treatment was done when the primary whitefly 

focus was detected: the presence of individual imago was noted on registration plants, 

larvae – not revealed. In Fig. 1 the dynamics of pest number is presented. 

The pest larvae occurrence on a control plot was noted 2 weeks earlier than in a 

variant with biological product application. Within the next four weeks the average 

whitefly number in both variants was at identical level. Later on in a control variant 

there was a sharp increase within a month (from 1 to 17 larvae per one leaf). In the experimental 

variant on 42 and 68 day from the experiment beginning there were repeated 

250

treatments using the biological preparation. The maximum pest number value did not 

exceeded 3 indiv./leaf (what was 6 times lower in comparison with the control). At the 

moment of researches termination (in 11 weeks after the experiment started) the larvae 

number has made, on the average, 10 indiv./leaf, in the experiment – 2 indiv./leaf. 

Fig. 1. Influence of Pecilomicine-B application on greenhouse whitefly 

Trialeurodes vaporariorum West. larvae number 

(greenhouse farm “DorOrs”, 2002 m., 

tomato ‘Raissa’ F 1 

, small-volume technology) 

1 pav. Pecilomicine-B įtaka šiltadaržinių baltasparnių 

(Trialeurodes vaporariorum Wеst.) 

lervų skaičiui (šiltnamių ūkis “DorOrs”, 2002 m., pomidorai 

‘Raissa’ F 1 

, mažagabaritė technologija) 

Researches in the greenhouse farm “DorOrs” were continued in 2003 on tomato 

crop ‘Barcelona’ F 1 

. For the first time Pecilomicine-B was applied on June 21 at pest 

larvae number 20 indiv./25 registration leaves. The subsequent treatments were done 

on June 30, August 4, September 11. The biological efficiency of a preparation has 

made from 60 to 88.0 %. In the control variant during the experiment whitefly larvae 

number has increased from 10 to 228 indiv./25 registration leaves. 

While carrying out the researches in 2003 in the greenhouse plant “Oziorny” on 

tomato crop (‘Blagovest’ F 1 

, soil ground) the first treatment was done when the pest 

number in the colonization focuses has made 23–27 larvae/25 leaves. On the whole, 

there were 4 treatments with 7 days interval. Pecilomicine–B biological efficiency is 

presented in Table 1. 

251

Таble 1. Pecilomicine-B influence on greenhouse whitefly number (tomato 

‘Blagovest’ F 1 

, soil ground, greenhouse farm “Оziorny”, 2003) 

1 lentelė. Pecilomicine-B įtaka šiltadaržinių baltasparnių gausumui (pomidorai ‘Blagovest’ 

F 1 

, dirvožemis, šiltnamių ūkis „Оziorny“, 2003 m.) 

Whitefly larvae 

number on 25 leaves 

Greenhouse whitefly larvae number on 

25 leaves / Biological efficiency 

before treatment Šiltadaržinių baltasparnių lervų skaičius ant 

Variant 

Šiltadaržinių baltasparnių 

lervų skaičius days from the start of the experiment 

25 lapų / Biologinis efektyvumas (%) 

Variantas 

ant 25 lapų prieš dienos nuo bandymo pradžios 

apdorojimą 5 12 19 26 

Pecilomicine-B, PS 27 90/42.4 145/68.5 198/77.1 356/63.7 

Control (without treatment) 23 133 391 736 835 

Kontrolė (be apdorojimo) 

LSD 05 

/ R 05 

2.6 6.7 12.2 14.5 

In 2003 in greenhouses of the Republican Ecological Educational Center there 

was an estimation of efficiency of Pecilomicine-B against greenhouse whitefly on 

cucumber, TSHA 14-27 F 1 

(soil ground). The initial pest number has made 82 larvae 

per 25 registration leaves. There were 4 treatments by a preparation in the course of 

24 days. Control variant – without treatment. Data on the biological efficiency of a 

preparation are presented in Table 2. 

Таble 2. Biological efficiency of Pecilomicine-B against greenhouse whitefly, 

(cucumber, TSHA 14-27 F 1 

, soil ground, Republican Ecological Educational 

Center, 2003) 

2 lentelė. Pecilomicine-B biologinis efektyvumas nuo šiltadaržinių baltasparnių (agurkai, 

TSHA 14-27 F 1 

, dirvožemis, Respublikinis ekologijos švietimo centras, 2003 m.) 

Pecilomicine-B, PS biological efficiency 

Pecilomicine-B biologinis efektyvumas (%) 

Days from the start of the experiment 

Dienos nuo bandymo pradžios 

3 6 9 13 16 24 

31.9 27.4 45.0 34.9 68.0 69.4 

In 2004 there were trials on the evaluation of the efficiency of the preparation 

Pecilomicine-B application against cucumber midge. Researches were carried out 

in greenhouse farm “Zhdanovichy” by cucumber growing using small-volume hydroponics 

on rockwool (‘Mystica’ F 1 

) and in the greenhouse farm “Oziorny” under 

soil ground conditions (‘Turnir’ F 1 

). According to the results of records concerning 

cucumber midge number within 3–4 weeks after treatment, there was its stabilization 

(farm “Oziorny”) or a decrease (farm “Zhdanovichy”) (Table 3). Afterwards, there was 

an increase in imago number what was connected with the flight of a new phytophage 

generation. In control variant, the pest number on 14th day after treatment increased 

2.5 times, on 21st day – 8 times. 

252

Таble 3. Biological efficiency of Pecilomicine-B against cucumber midge 

(Bradysia brunnipes Mg.) 

3 lentelė. Pecilomicine-B biologinis efektyvumas nuo uodelių (Bradysia brunnipes 

Mg.) 

Imago number 

Pest number 

Suaugėlių skaičius (сm 2 ) 

before treatment, 


Variant 

imago 


Variantas 

Kenkėjų skaičius 

days after treatment 

prieš apdorojimą, 

dienos po apdorojimo 

suaugėliai (cm 2 ) 

7 14 21 28 

Cucumber ‘Mystica’ F 1 

, rock wool, greenhouse farm “Zhdanovichy”, 2004 

Agurkai ‘Mystica’ F 1 

, akmens vata, šiltnamių ūkis “Zhdanovichy”, 2004 m. 

Pecilomicine-B, PS (40 kg/hа) 0.12 0.06 

50.0 

0.02 

83.3 

0.04 

66.6 

0.03 

75.0 

Cucumber ‘Тurnir’ F 1 

, soil ground, greenhouse farm “Оziorny”, 2004 

Agurkai ‘Тurnir F1’, dirvožemis, šiltnamių ūkis “Оziorny”, 2004 

Pecilomicine-B, PS (40 kg/ha) 0.22 0.21 0.31 0.37 - 

35.8 40.0 71.5 

Control (water irrigation) 

Kontrolė (drėkinimas vandeniu) 

0.21 0.28 0.53 1.72 - 

LSD 05 

/ R 05 

- - - 0.48 - 

To study the influence of Pecilomicine-B application on encarsia there were the 

researches done under greenhouse conditions (greenhouse farm “DorOrs”) on tomato 


. In all the area of greenhouse in the focuses of whitefly occurrence encarsia 

was released at the rate predator: victim = 1 : 10. In 12 days after the entomophage 

release, greenhouse whiteflies (110–130 indiv./leaf) were treated with Pecilomicine-B 

(1 %). Pecilomicine-B action on encarsia was estimated by a quantity of whitefly 

infected larvae and flying out encarsia imago, presented in Table 4. 

Таble 4. Influence of Pecilomicine-B on encarsia (Encarsia formosa Gahan.) 

survival (tomato ‘Raissa’ F 1 

, greenhouse farm “DorOrs”, 2005) 

4 lentelė. Pecilomicine-B įtaka enkarzijų (Encarsia formosa Gahan.) išlikimui (pomidorai 


, šiltnamių ūkis “DorOrs”, 2005) 

Variant 

Variantas 

Average number of the 

infected larvae in the 

sample (indiv.) 

Vidutinis užkrėstų lervų 

skaičius pavyzdyje, individai 


flying out encarsia imago 

(indiv.) 

Vidutinis išskridusių 

enkarzijų suaugėlių skaičius, 

individai 

Survival of 

encarsia imago 

Enkarzijų suaugėlių 

išlikimo 

(%) 

Pecilomicine-B, PS 157 142.7 91.7 

Control (without treatment) 167 152.5 87.4 

Kontrolė (be apdorojimo) 

LSD 05 

/ R 05 

7.8 

253

The next experiment was carried out for studying the influence of Pecilomicine-B 

on phytoseiulus survival rate and its ability to reproduce (greenhouse farm 

“Zhdanovichy”, 2006). Soybean plants with the entomophage culture were treated with 

preparation (1 %). Phytoseiulus number in the control (without treatment) increased 

2.3 times (up to 134 individuals), on Pecilomicine-B treated plot – 2.6 times (up to 

146) for 12 days (Fig. 2). 

Fig. 2. Influence of Pecilomicine-B on phytoseiulus Phytoseuilus persimilis Ath. 

(greenhouse farm “Zhdanovichy”, soybean, 2006) 

2 pav. Pecilomicine-B įtaka fitoseiliams (Phytoseiulus persimilis Ath.) 

(šiltnamių ūkis “Zhdanovichy”, sojų pupelės, 2006 m.) 

Discussion. There are recommendations on the expediency of pest economic 

thresholds of harmfulness to use as a reference point for plant protection products 

application (including the biological ones) against it. So, according to the data presented 

by V. A. Pavlyushin and his co-workers (2001), for greenhouse whitefly it 

makes 40 individuals of different development stages per cucumber leaf and 10 individuals 

per tomato leaf. At the same time there is an opinion that the use of indicators 

of thresholds of harmfulness in the greenhouses where action of limiting factors 

of the environment on dynamics of phytophage populations number is considerably 

weakened is inadequate to situation. Martens (1993), Osborne, Landa (1992), Faria 

et al. (2001) recommend for increasing the efficiency of P. fumosoroseus application 

to carry out mycoinsecticide treatments during occurrence of early stages of tobacco 

whitefly development (from the moment of the first whitefly imago appearance and 

not more than 1 imago per plant). The results of our researches showed that the most 

effective Pecilomicine-B action (the biological efficiency up to 100 %) on the phytophage 

is observed in case of consecutive application of a preparation beginning at 

the initial stage of greenhouse crop plantings colonization by whitefly (right after the 

first imago occurrence in plants). Therefore, in pest population the necessary “patho- 

254

genic press” is initially created. Thus, a full suppression of population development 

was noticed within 14–30 days, while in the control variant larvae number increased. 

Similar dependence was observed in the experiments on the efficiency of Pecilomicine-B 

on cucumber midge. As it is seen in Table 3, higher biological efficiency of 

a preparation (83.3 %) has been noted in case smaller initial cucumber midge number 

(0.12 imago/сm 2 ). Nevertheless, in case of significant initial pest larvae number 

(whitefly – 20–82 larvae per 25 registration leaves, cucumber midge – 0.22 imago/ 

сm 2 ) the application of a preparation has appeared sufficient (60–88 % for whitefly, 

35.0–71.5 % for cucumber midge) to lower considerably speed of pest population 

increase. 

Availability of insecticidal properties in entomopathogenic fungi assumes a possibility 

of their influence not only on target objects, but also on pest entomophages 

and parasites. In this connection a number of authors (Павлюшин, Агансонова, 1994; 

Павлюшин, 1996) consider a combination of mycoinsecticides and useful insects in 

the protection systems problematic. There are messages on a competition between 

entomophage and pathogen in relation to host, resulting in efficiency decrease of both 

agents (Соловей, Забудская, 1985). Nevertheless, the results of many researches testify 

to the absence of negative action of fungi on certain useful insect species (Михневич, 

Климпиня, 1988; Roditakis et al., 2001; Alma et al., 2007). Also the increase of entomophage 

activity with the combined use of biological preparations is marked, caused 

by immunity decrease in noxious insects to parasites as a result of their organism weakening 

under the pathogen influence (Исаева, 1976). Our evaluation of Pecilomicine-B 

action on arthropods at combined application under greenhouse conditions and also 

the results of our preliminary laboratory and vegetative experiments (Прищепа et 

al., 2007) testify the absence of the negative preparation action on survival, ability to 

reproduce, predatory and parasitic activity of phytoseiulus and encarsia. 

Conclusions. 1. It is the most expedient to start the application of Pecilomicine-B 

at a stage of primary colonization of greenhouse crop plantings by phytophages: 

in case with greenhouse whitefly – at occurrence of the first imago on plant leaves by 

carrying out 2 treatments at 7–14 days interval. Later on, Pecilomicine-B (1 % concentration) 

is applied up to 4 times considering the dynamics of greenhouse whitefly 

number, namely in case of its stable increase. It allows avoiding within 1.5–2 months 

the intensive increase of phytophage number without chemical plant protection products 

application. Against cucumber midge it is expedient to apply Pecilomicine-B 

(4 % concentration up to 2 times at 23–27 days interval) with the beginning of mass 

pest imago flight. 

2. Application of Pecilomicine-B in greenhouses in a combination with the release 

of phytoseiulus and encarsia (at 7–14 days interval) does not render a negative 

influence on survival, reproduction, parasitic and predatory activity of entomophages 

and does not decrease the efficiency of their action. 

3. Pecilomicine-B application according to the above-mentioned parameters allows 

constraining phytophage population number at economically imperceptible level. 

Gauta 2009 0630 


255

References 

1. Alma C. R., Goetell M. S., Roitberg B. D., Gillespie D. R. 2007. Combined 

effect of the enthomopathogenic fungus, Paecilomyces fumosoroseus Apopka- 

97, and the generalist predator, Dicyphus Hesperus, on the whitefly populations. 

BioControl, 52: 669–681. 

2. Faria M., Wraight S. P., Naranjo S. E., Ellsworth P. C. 2001. Biological control 

of Bemisia tabaci with fungi. Crop Protection, 20(9): 767–778. 

3. Lacey L. A., Frutos R., Kaya H. K., Vails P. 2001. Insect pathogens as biological 

control agents: do they have future Biological-Control, 21: 230–248. 

4. Martens J. A. 1993. Fungus and new crops build business in Florida. Grower 

Talks, 56(11): 34–41. 

5. Nigroho I., Ibrahim Y. 2004. Laboratory bioassay of some enthomopathogenic 

fungi against broad mite (Polyfagotarsonemus latus Bank.). Journal of 

Agriculture and Biology, 6(2): 223–225. 

6. Osborne L. S., Landa Z. 1992. Biological control of whiteflies with entomopathogenic 

fungi. Florida Entomologist, 75(4): 456–471. 

7. Pavlyushin V. A. 1996. Effect of entomopathogenic fungi on entomophagous 

arthropods. IOBC/WPRS Bulletin, 19(9): 247–249. 

8. Poprawski T. J., Jones W. J. 2000. Host plant effects on activity of mitosporic 

fungi Beauveria bassiana and Paecilomyces fumoso-roseus against two populations 

of Bemisia whiteflies (Homoptera: Aleyrodidae). Mycopathologia, 151: 

11–20. 

9. Roditakis N. E., Charnley K., Roditakis E. N. 2001. The green aphids Myzus persicae 

and its control with combined use of entomopathogenic fungus Verticillium 

lecanii and parasitoid Aphidius colemani under greenhouse conditions on sweet 

peppers in Crete. Abstracts of 8 th European meeting of the IOBC/WPRS Working 

Group “Insect Pathogens and Insect Parasitic Nematodes” (May–June 2001), 

Athens. 

10. Shi W.-B., Feng M.-G. 2009. Effect of fungal infection on reproductive potential 

and survival time of Tetranychus urticae (Acari: Tetranychidae). Expermental 

and Applied Acarology, 229–237. 

11. Thungrabeab M., Blaeser P., Sengonca C. 2006. Possibilities for biocontrol of 

the onion thrips Thrips tabaci Lindeman (Thys., Thripidae) using different entomopathogenic 

fungi from Thailand. Mitteilungen der Deutschen Gesellschaft 

für allgemeine und angewandte Entomologie, 15: 229–304. 

12. Wraight S. P., Carruthers R. I., Jaronski S. T., Bradley C. A. et al. 2000. Evaluation 

of the entomopathogenic fungi Beauveria bassiana and Paecilomyces fumoso-roseus 

for microbial control of the silverleaf whitefly Bemisia argentifolii. 

Biological control, 17: 203–217. 

13. Ахатов А. К. (ред.) 2004. Вредители тепличных и оранжерейных растений 

(морфология, образ жизни, вредоносность, борьба). Товарищество научных 

изданий КМК. Москва. 

256

14. Ижевский С. С. (ред.) 1999. Защита тепличных и оранжерейных растений от 

вредителей: Справочник (определение, видов, методы выявления и учета, 

биология и морфология, вредоносность, борьба). KMK Scientific press Ltd. 

Москва. 

15. Исаева Л. И. 1976. Использование биологического и новых методов защиты 

растений в интегрированных программах. ВНИИТЭИСХ. Москва. 

16. Михневич О. Ч., Климпиня А. Е. 1988. Оценка влияния энтомопатогенных 

грибов на коровку Cycloneda limbifer Casey. Биологическая регуляция 

численности вредных членистоногих. Зинатне. Рига, 56–67. 

17. Павлюшин В. А., Агансонова Н. Е. 1994. Совместимость энкарзии и 

алейцида в борьбе с оранжерейной белокрылкой на огурцах в теплицах. 

Экологически безопасные и безпестицидные технологии получения 

растениеводческой продукции: материалы Всерос. науч.-произв. совещ. 

(Краснодар, авг. 1994 г.). Пущино, 2: 165–168. 

18. Павлюшин В. А. (ред.) 2001. Система биологической защиты овощных 

культур от вредителей и болезней в теплицах. Всероссийский НИИ защиты 

растений (ВИЗР). Санкт-Петербург, 72. 

19. Прищепа Л. И. 2005. Опыт применения биоинсектицида пециломицин-Б в 

защите овощных культур от тепличной белокрылки. Ахова раслiн, 42 (5): 

28–30. 

20. Прищепа Л. И., Микульская Н. И., Войтка Д. В. 2008. Методические 

указания по проведению регистрационных испытаний биопрепаратов 

для защиты растений от вредителей и болезней. МОУП «Несвижская 

укрупненная типография им. С. Будного». Несвиж. 

21. Прищепа Л. И., Янковская Е. Н. 2008. Штамм Paecilomyces fumosoroseus 

БИМ—F-328Д для производства энтомопатогенного препарата для защиты 

растений: пат. №10813 Республики Беларусь, МПК А 01 N 63/04. – опубл. 

30.06.08. Афiцыйны бюл. Нац. цэнтр iнтэлектуальн. уласнасцi, 3: 45. 

22. Прищепа Л. И., Янковская Е. Н., Гарко Л. С. 2007. Перспективы совместного 

применения Paecilomyces fumosoroseus (Wize) Brown et Smith и энтомофагов 

для защиты томата от вредителей. Сб. науч. тр. РУП «Институт защиты 

растений», 31: 325–335. 

23. Соловей Е. Ф., Забудская И. А. 1985. Опыт совместного применения энкарзии 

(Encarsia formosa Gah.) и грибов рода Aschersonia и Verticillium lecanii 

Zimm. в борьбе с тепличной белокрылкой на огурцах. Интегрированная 

защита овощных и плодовых культур. Штииница. Кишинев, 28–33. 

24. Янковская Е. Н. 2006. Разработка технологии применения препарата 

пециломицин-Б для защиты огурца от огуречного комарика. Сб. научн. тр. 

РУП «Институт защиты растений», 30(2): 206–213. 

25. Янковская Е. Н., Прищепа Л. И. 2008. Исследование биотехнологических 

параметров получения биоинсектицидного препарата пециломицин-Б. 

Сб.научн.тр. РУП “Институт защиты растений”, 32: 368–377. 

257


Biologinio insekticido Pecilomicine-B poveikis šiltnamio kenkėjams 

A. Yankouskaya 

Santrauka 

Šiltnamio sąlygomis buvo įvertinta kai kurių technologinių parametrų (terminų, apdorojimų 

skaičiaus ir intervalų tarp jų) įtaka biologiniam bioinsekticido Pecilomicine-B poveikiui 

šiltadaržinio baltasparniui (Trialeurodes vaporariorum Wеstw.), uodeliams (Bradysia brunnipes 

Mg.), entomofagams – enkarzijoms (Encarsia formosa Gahan.) ir fitoseiliams (Phytoseiulus persimilis 

Ath.). Nustatyta, kad efektyviausia naudoti Pecilomicine-B, vos tik ant šiltnamio augalų 

pasirodo fitofagai: šiltadaržinio baltasparnio atveju – kai ant lapų pasirodo pirmieji suaugėliai, 

kas 7–14 dienų apdorojant juos du kartus. Vėliau 1 % koncentracijos Pecilomicine-B naudojamas 

iki 4 kartų, priklausomai nuo šiltadaržinio baltasparnio populiacijos gausumo (jeigu jų gausumas 

didėja). Tai leidžia 1,5–2 mėnesius be cheminių augalų apsaugos priemonių išlaikyti fitofagų 

populiaciją ekonomiškai nežalingame lygyje. Nuo uodelių šiuo preparatu (4 % koncentracijos) 

reikėtų apdoroti iki 2 kartų kas 23–27 dienos suaugėlių masinio skraidymo pradžioje. Kai 

Pecilomicine-B naudojamas šiltnamio sąlygomis (kas 7–14 dienų) kartu su fitoseiliais ir enkarzijomis, 

jis nedaro neigiamos įtakos entomofagų išlikimui, dauginimuisi, parazitinei bei grobuoniškai 

veiklai ir nesumažina jų efektyvumo. Naudojant Pecilomicine-B pagal minėtuosius technologinius 

parametrus, galima išlaikyti fitofagų populiacijas ekonomiškai nežalingame lygyje. 

Reikšminiai žodžiai: bioinsekticidų efektyvumas, bioinsekticidų įtaka entomofagams, 

Bradysia brunnipes, Cucumis sativus, Encarsia formosa, Lycopersicon esculentum, 

Paecilomyces fumosoroseus, Pecilomicine-B, Phytoseiulus persimilis, Trialeurodes vaporariorum. 

258




Optimization of time and expediency of 

Incurvaria capitella Cl. number regulation 

Svetlana Yarchakovskaya, Natallia Kaltun 

RUC Institute of plant protection p. Priluki, Мinsk region, Belarus, 


Investigations were conducted in black currant plantations (Minsk district) in 1994–2008. 

The objects of researches were black currant cultivars growing in Belarus and currant bud 

moth (Lampronia (Incurvaria) capitella Cl.). 

The objective of researches was to develop currant bud moth monitoring system in 

black currant plantations based on phenological forecast of pest development. There was applied 

original synthetic phytophage sex pheromone considering a degree of different currant 

cultivar damage. 

An algorithm of phenological forecast of currant bud moth development was worked out. 

It gave an opportunity to determine beforehand the optimum time of registering of caterpillars 

leaving their wintering places, imago flight dynamics and carrying out a complex of chemical 

and agrotechnical measures. The attractiveness of the original synthetic phytophage sex 

pheromones was studied. It was determined that as a dispenser for the synthetic sex pheromone 

it is preferable to use the insulin cork or rubber black tube with a. i. content 1 mg/dispenser. 

It is determined that mid-early cultivars (‘Minay Shmyrev’, ‘Partisanka’) under conditions of 

Belarus are damaged by black currant moth much stronger than the mid and mid-late ones. 

The least pest-damaged cultivar is mid-late ‘Zolushka’. 

Key words: attractiveness, black currant, cultivar damage, development forecast, 

Incurvaria capitella, monitoring, pheromone. 

Introduction. Currant bud moth Lampronia (Incurvaria capitella) Cl. is a constant 

representative of currant fauna in all the regions of this crop cultivation. The 

phytophage damages both black and red currant, however, prefers a black one (Labanowska, 

2003). Black currant bud damage do not using the protective measures 

in years of mass pest development can reach 80–100 %, what practically leads to 

full crop loss. Basing on literary data, the most strongly damaged are early cultivars 

(Савздарг, 1960). Some features of the phytophage development complicate 

its monitoring, abundance and spread restriction. Already during black currant bud 

swelling, which depending on weather conditions can be observed even in February, 

the first wintering caterpillars leave the shelter places and bite into currant buds. To 

catch visually the beginning of pest appearance from wintering places it is necessary 

259

to carry out periodic (in 3–4 days) observations in plantations since February and 

prior to the beginning of currant bud breaking. This is rather laborious and expensive 

(Крикунова и др., 2007). It shows an urgency of investigations, which would improve 

the methods of doing records and optimize the terms of protective measures. 

Being in buds the caterpillars feed prior to the beginning of black currant blossoming. 

The pest pupation starts in the top soil layer during the period of floral racemes 

advancing before mass black currant blossoming. The butterfly flight starts in May 

and coincides with black currant blossoming termination. As currant bud moth imago 

are active for a very short period of time, that is from 6.30 to 9.30, certain difficulties 

monitoring their abundance and timely revealing the focuses of their distribution 

are created. The female lays eggs in pulp of green berries. The hatched caterpillars 

within several days eat seeds of berries, and then disappear in shelters for wintering 

(Ярчаковская, 2000). 

So, high phytophage harmfulness, the latent way of life of a harmful stage, low 

and short imago activity determine the expediency of carrying out the researches on 

working out the methods and techniques of its monitoring on different by maturation 

cultivars. 

For terms optimization and the expediency of realization of these or the other 

actions for plant protection by working out the modern strategies of insect pest population 

control a great attention is given to the development and use of phenological 

forecasts of phytophage development and also studying host-plant cultivars infection. 

The use of synthetic sexual pest pheromones (SSP) for their number monitoring is 

also rather perspective. 

In connection with the above-stated, the purpose of our researches was working 

out the algorithm of phenological forecast of currant bud moth development for optimization 

the terms of conducting pest abundance assessments and protective measures, 

creation of the synthetic sexual phytophage pheromone for timely revealing the 

focuses of its occurrence, an estimation of various currant cultivars infestation by the 

phytophage for determining the expediency of carrying out sprayings. 

Objects, methods and conditions. The observations of currant bud moth development 

phenology, collection of biological material, field and farming trials were 

conducted in black currant plantations of fruit-growing farms, Minsk district (“Zubki”, 

“Agroconcern Kletsky”, “Uzdensky”), in 1994–2006. The observations on the 

assessment of black currant cultivars phytophage infestation were started on an experimental 

plot of Fruit-growing Institute (Minsk district) in 2003–2004. The attractiveness 

of local samples of currant bud moth synthetic sex pheromones was evaluated in 

fruit-growing plantations of the farm “Zubki” (Minsk district) in 2007 and 2008. 

The objects of researches were the regionalized in Belarus black currant cultivars 

(‘Мinaj Shmyrev’, ‘Partizanka’, ‘Kаtyusha’, ‘Оdzhebin’, ‘Каntate’, ‘Belarusskaya 

sladkaya’) and currant bud moth (Lampronia (Incurvaria) capitella Cl.). 

Investigation of migration dynamics of currant bud moth wintered caterpillars 

from wintering places to buds, their harmful activity period, dynamics of leaving 

for pupation was done by daily observations of ten registered black currant bushes 

colonized by moth. From the moment of buds swelling 50 buds were inspected on 

260

tops of branches of each registered bush and the number of pest caterpillars feeding 

in them was recorded. The observations of caterpillar development were done by 

their individual maintenance on currant branches free of other pests (1 caterpillar on 

a branch). The duration of pupae development was studied by their individual development 

in not less than 20 test tubes with soil. An establishment of butterflies fly out 

dynamics was done in entomological insect cages. Insect cage size was 30 x 30 x 50 

cm and it was in a form of a wooden frame fitted from three sides by a kapron sieve 

and from the fourth side glazed by sliding glass, cage’s bottom and top – from wood. 

The cage’s bottom was filled with 5–6 cm volume sifted soil layer. Black currant branches 

colonized by bud moth last age caterpillars (400–500 individuals) were placed 

in each cage. Every 2 days from the moment of fly out beginning butterfly number 

was recorded. Observations on imago life duration, female fecundity were done on 

currant branches under gauze insulators put on a wire frame, where the just flied out 

individuals were placed (15 insulators having 1 female and 2 male in an insulator). 

The insulators were daily inspected. After butterflies were killed by opening berries, 

the number of eggs laid by a female was recorded. Dynamics of caterpillars hatching, 

their feeding duration in berries and periods of diapause leaving were determined by 

periodic (every three days from the beginning of egg laying) 100 berries opening taken 

from the pest-infested plantation. 

The evaluation of black currant cultivar damage by bud moth was done by inspecting 

50 buds on each of 10 registration bushes of each cultivar and recording the 

pest-damaged buds after the phytophage brought full damage. 

The itinerary inspections and estimation of currant plantation phytosanitary condition 

were done by the standard methods (Алехин и др., 1988; Грин и др., 1996). 

Systematization, generalization and statistic processing of the collected material 

for creating the algorithm of phenological forecast of currant bud moth development 

were done by the method of correlation and regression analysis (Zar, 1996; Уланова, 

Забелин, 1990). Relations between constant indicators and the application of the investigated 

relationships for the forecasting equations were done by multiple regression 

analysis method. Reliability of the developed evaluations was verified by F – Fisher’s 

criterion and t – Student’s criterion, supposing 95 % significance level. The meteorological 

data necessary for the calculations have been obtained from agrometeorological 

stations, located close to the place of the researches. 

For doing a primary estimation of attractiveness of currant moth synthetic sexual 

pheromone (SSP) samples before black currant blossoming the plots were selected 

with a high pest number. During blossoming on selected plots pheromone-glue traps 

of the type Atrakon-A with various samples of synthetic sex pheromones provided by 

the employees of Elementary Synthesis Scientific-Research Laboratory of Belarussian 

State University were hung out. The experiments were accomplished in 5–7 repetitions 

(1 repetition – 1 trap). Traps were numbered and hung out in the top part of black 

currant bush. The traps were located along the plot in a randomized way in a distance 

of not less than 30 m from each other and from planting edge. The trap records were 

done regularly every 7 days. The caught butterflies were recorded and taken away 

from a sticky surface. For experiments a sticky mass “Vinilon” was used. Glutinous 

261

loose leaves in traps were replaced as required. Attractiveness of all the presented SSP 

samples was estimated by average number of caught butterflies. 

Results. Based on systematization and statistic analysis of results of 13 summer 

observations on bud moth biology and phenology, the algorithm of pest development 

forecast, i.e., periods of phytosanitary situation monitoring in black currant plantations, 

was developed. 

By working out logic forecasting model of currant bud moth development the 

influence of different environmental factors on speed of the pest development (relative 

air humidity, rainfall sum, photoperiod duration, and minimum, maximum and 

average daily air temperature) was evaluated. It is determined that the main predictors 

of forecast are air temperature (minimum, maximum and average daily), rainfall sum 

and also photoperiod duration. 

Mathematical forecasting model of currant bud moth development is expressed by 

a series of multifactorial and monofactorial linear and polynominal regression equations 

depicting quantitative characteristics of relations of forecasted phenomena and 

the environmental conditions. The close relations between the investigated phenomena 

were evaluated as reliable, if correlation coefficients (r y, x 

and R y, x 1, x 2…xi 

) were equal to 

0.7 and more. By predictors selection for mathematical forecasting model development 

the chosen indicators should not correlate among themselves (r x, x1 

< 0.4). 

It is determined that a primary point for phenological forecast development is a 

date of maximum temperatures transition through +5 °С. 

It is established that the forecast predictors of the phytophage development beginning 

(1-st stage of the forecast) are maximum air temperatures and photoperiod 

duration. Time of bud moth caterpillars leaving from wintering places is caused by the 

accumulation of certain amount of positive temperatures. It is calculated that caterpillars 

start to leave their wintering places if for each 100 minutes of a light day, on the average, 

1 °C warmth comes in April, 1.2–1.5 °C – in March and 2–2.5 °C – in February. 

So, the earlier (at much shorter light day) the higher maximum air temperatures are 

necessary for the pest to start development. 

The pest caterpillars start leaving their wintering places if at the end of February 

a light day length is 600 minutes and daily air temperatures reach +15 °C, in the 

beginning of March (about 650 minutes) – +13 °С, in the beginning of April (about 

750 minutes) – +9 °С. 

The calculated forecasting equation depicting the interdependence of maximum 

air temperature, providing the beginning of pest development and photoperiod duration 

looks like parabola with 97 % level of correlation dependence with an error of 

determination coefficient 0.8 % and registers as follows: 

Y = 0.0001097 х 2 - 0.1896414 х + 90.6988633, D = 0.97 (1) 

Using the calculated regression equation, it is possible to define the exact date 

of bud moth leaving the wintering places, i. e., the optimum time of doing the pest 

records and sprayings against the phytophage. 

It is also established that the duration of currant bud moth caterpillars feeding in 

buds (2-nd stage of the forecast) is determined by air temperature parameters. For all 

years of observations depending on developing weather conditions the pest caterpillars, 

262

which have left their wintering places, have been eating the broken buds from 20 to 50 

days. The main factors influencing the period of currant moth feeding (Y 1 

) are average 

daily temperature (х 1 

) and the sum of minus air temperatures (х 2 

) for the feeding period. 

The equation reflecting quantitative interrelation of the above-mentioned parameters 

registers as follows: 

У 1 

= 23.88 - 0.422 х 1 

+ 0.496 х 2 , D = 0.70 (2) 

The obtained equation allows predicting terms of pest feeding and the beginning 

of caterpillars leaving for pupation, i. e., time of inter-row hoeing, whenever possible 

close to bushes for decreasing the pupae number. 

The period of currant bud moth development (3-rd stage of forecast) and terms of 

oviposition by females (4-th stage of forecast) are defined by air temperature indicators 

and moisture content. 

For all the years of observations, depending on weather conditions currant bud 

moth pupae developed from 12 to 36 days. The main predictors influencing the duration 

of their development (Y 2 

) are maximum air temperature (x) and the rainfall sum (х 3 

) for 

the period of pupae development. The equation depicting the quantitative interrelation 

of the above-mentioned indicators registers as follows: 

Y 2 

= 65.1 - 2.29 х + 0.05 х 3 

, D = 0.71 (3) 

Using the obtained equation, it is possible to predict terms of pest butterflies flight 

beginning and, hence, terms of hanging out pheromone-glue traps for the purpose of 

monitoring phytosanitary condition of black currant plantations and revealing the 

focuses of its spread. 

It is also established that from the beginning of currant bud moth flight up to 

the start of laying eggs by females can pass from 2 to 12 days, what also depends on 

prevailing weather conditions. However, in this case the basic defining factor is the 

minimum air temperature (x 4 

) and also as in the 3-rd stage, rainfall sum (х 3 

) for summer 

period. The quantitative interrelation of the above-named indicators is defined 

by equation Y 3 

= 17.0 - 1.12 х 4 

+ 0.07 х 3 

, D = 0.73 (4), i. е. the higher minimum air 

temperature and less rainfall, the quicker females start laying eggs. 

The basic indicators defining the duration of egg development period (Y 4 

) and 

terms of the phytophage larvae leaving for wintering in shelters (Y 5 

) also are the 

minimum air temperature (х 4 

) and rainfall sum (х 4 

). For all years of inspections the 

duration of egg development period varied from 8 to 30 days and the duration of hatched 

from eggs caterpillars before their leaving for wintering in shelters – from 4 to 

14 days, what was defined first of all by rainfall sum and minimum air temperatures. 

The regression equations reflecting the quantitative interrelation of the above-named 

indicators register as follows: 

Y 4 

= 7.3 - 0.44 х 4 

+ 0.11 х 3 

, D = 0.67 (5) and 

Y 5 

= 4.2 - 0.41 х 4 

+ 0.05 х 3 

, D = 0.69 (6). 

Thus, based on done researches an algorithm of phenological forecast of 6 stages 

of currant bud moth development was developed beginning from terms of caterpillar 

appearance from wintering places up to the beginning of their leaving in shelters. The 

consecutive use of the calculated equations allows to define beforehand the optimum 

terms of number records of caterpillars appearing from wintering places, imago flight 

263

dynamics and realization of a complex of chemical (against caterpillars) and agrotechnical 

(against pupae) measures. 

For timely revealing of I. сapitella focuses the workers of the Belarussian Institute 

of Plant Protection together with the employees of the scientific-research laboratory 

of elementary synthesis of Belarus State University (BSU) carried out the researches 

on working out the pest synthetic sex pheromone (SSP). For primary evaluation of the 

pest SSP the workers of the BSU have manufactured 14 experimental samples under 

the conventional name “Lavabat”, differing by the active ingredient content (0.1, 0.2, 

1 mg/dispenser) and the dispenser type (black and white rubber tubes, blue sponge, 

insulin cork). Among all the tested samples the attractiveness in relation to currant bud 

moth males have shown 8. It is determined that as a dispenser for currant bud moth 

SSP it is more preferable to use the insulin cork or rubber black tube with a. i. amount 

1 mg/dispenser. Using these samples for the period of pest flight 21.05–17.06 has 

resulted in 11.0–60.6 butterflies caught (Table 1). 

Таble 1. Attractiveness of SSP experimental samples for currant bud moth 

(Lampronia (Incurvaria) capitella Cl.), Мinsk district, 2007–2008 (currant age 

6–7 years) 

1 lentelė. Eksperimentinių serbentinės kandies (Lampronia (Incurvaria) capitella Cl.) sintetinio 

lytinio feromono pavyzdžių patrauklumas, Minsko regionas, 2007–2008 (serbentų 

amžius 6–7 metai) 

Experimental 

sample 

Eksperimentinis 

pavyzdys 

Active ingredient 

content 

(mg/dispenser) 

Veikliosios 

medžiagos kiekis, 

mg/dalytuvas 

Dispenser type 

Dalytuvo tipas 

Butterflies caught during 

flight period 

(the average per one trap) 

Drugiai, pagauti vieno 

skridimo laikotarpiu, vidutiniškai 

vienose gaudyklėse 

10.4 

Lavabat Т 1 1.5 сm of white rubber tube 

0.1 1,5 cm baltos guminės žarnelės 

1.8 

Lavabat D 1 4.0 

0.1 1.2 

Lavabat ТG 1 Blue sponge circle 

5.0 

Mėlynos kempinės ratas 

Lavabat TP 1 Insulin cork 

60.6 

Lavabat P 0.2 Insulino kamštis 

11.0 

Lavabat ТR 0.2 2 сm of black rubber tube 

2 cm juodos guminės žarnelės 

11.2 

To determine the expediency of conducting the protective measures, besides, 

timely revealing of the phytophage incidence focuses, its abundance assessments, it 

is also necessary to have the information concerning host plant cultivar infestation. 

For this purpose, the infestation studies on 7 black currant cultivars differing by maturation 

time were carried out in 2003–2004. Bud damage records were done after a 

full damage during buds advancing. 

264

Table 2. Black currant cultivars damaged by currant bud moth, Samokhvalovichi 

2 lentelė. Serbentinės kandies pažeistos juodųjų serbentų veislės, Samokhvalovichi 

Cultivar 

Veislė 

Maturation time 

Sunokimo laikas 

Bud damage 

Pumpurų pažeidimas (%) 

2003 2004 

26.2 24.2 

‘Мinay Shmyrev’ Mid-early 

Vidutiniškai ankstyva 

‘Partizanka’ 

Mid-early 

24.0 18.0 

Vidutiniškai ankstyva 

‘Каtyusha’ 

Mid-ripening 

10.5 8.2 

Vidutiniška 

‘Odzebin’ 

Mid-ripening 

10.3 7.9 

Vidutiniška 

‘Каntata’ 

Mid-ripening 

2.7 9.0 

Vidutiniška 

‘Belorusskaya sladkaya’ Mid-ripening 

1.8 4.0 

Vidutiniška 

‘Zolushka ’ 

Mid-late 

1.4 2.4 

Vidutiniškai vėlyva 

LSL 05 

/ R 05 

11.46 8.13 

It is determined that mid-early cultivars were damaged by currant bud moth much 

stronger than mid-ripening and mid-late. The mid-early ripening cultivars ‘Мinay 

Shmyrev’ and ‘Partizanka’ bud damage has reached 18–26 %, what is two and more 

times higher than the mid-ripening cultivars (Table 2). The least bud damage was 

noticed in mid-late cultivar ‘Zolushka’. 

Discussion. World literature analysis concerning the questions of time optimization 

on carrying out one or another measure in plant protection showed that the 

similar researches were carried out mainly in the direction of phenological forecasts 

of agricultural host plant development. In Bulgaria, Slovenia, Japan, USA, Canada 

different regression models of apple, pear, plum (Hricovsky et al., 1994; Jonaitis, 

1994; Kajfez-Bogataj, Bergant, 1998; Morgan, Solomon, 1993), citrus (Ono, Konno, 

1999), red currant (Вандова, 2000) development based on the use of quantitative and 

qualitative characteristics of agrometeorological parameters were worked out. Based 

on the results of researches, the main forecast predictors are air temperature and 

rainfall sum. In Russia the investigations are carried out in modeling direction and 

forecast of the main productive processes of grain, fodder, vegetable and technical 

crop field agrocoenosises ( Бородий, Зубков, 2001; Полуэктов, 2001). 

The data show that in all existing models a task of the qualitative relation establishment 

of the calendar time with the physiological one is solved by similar methods 

differing by some details. As a leading factor influencing the current speed of 

the studied object, in the developed models a sum of the effective and average daily 

air temperature is used. As an additional predictor, authors consider water regime or 

water-capacity (rainfall sum, relative air humidity and etc.). Nearly all the authors 

265

indicate that the duration of the light period is rather important to the individual object 

development, however, they consider this parameter as proceeding automatically in 

a certain location and not taken into account in the developed models. In the researches 

authors (Koltun, et al., 2003) determined that at initial stages of both the studied 

phytophage and black currant development, the light period duration as well as air 

temperature was the main predictor of the developed model of phenological forecast 

of currant bud moth Lampronia (Incurvaria capitella Cl.). 

Similar researches are done in Poland by Barbara Labanowska (2003), who 

indicated a close relation of caterpillars leaving the wintering places with daily air 

temperature and also the possibility of phytophage development beginning in February 

under air temperatures 10–15 °C and the necessity of carrying out 2–3 sprayings 

against the pest. Her recommended protective measures are determined basing on 

field records and observations. The first treatment should be done when the pest leaves 

the wintering places by pyrethroid group preparations. When the repeated sprayings 

are necessary against the larvae bitten into buds it is recommended to use contact 

and systemic preparations. However, in the indicated researches it is not mentioned 

clearly determined the most optimum time of carrying out the protective measures, 

what does not allow to provide with efficiency higher than 55–78 % even when such 

high-toxic synthetic pyrethroids as Decis and Fastac are used. Moreover, carrying out 

2–3 treatments against one pest makes protection significantly expensive, increases 

the energy resources, decreases the profitability of the crop growing and negatively 

affects the environmental safety. 

As a result of our researches a method of instrumental optimum time of spraying 

against the pest is proposed at the start of Incurvaria capitella Cl. caterpillars leaving 

their wintering places using the regression equation of dependence between maximum 

air temperature parameters and photoperiod duration. 

Conclusions. An algorithm of the pest Lampronia (Incurvaria) capitella Cl., development 

forecast an opportunity to determine beforehand the optimum time of doing 

records of caterpillars leaving their wintering places, imago flight dynamics and 

carrying out a complex of chemical (against caterpillars) and agrotechnical (against 

pupae) measures in black currant plantations. 

For timely revealing of I. capitella focuses distribution in black currant plantations 

the attractiveness and possibility of the phytophage original synthetic sex pheromones 

is studied. It is determined that as a dispenser for currant bud moth synthetic 

sex pheromone it is more preferable to use insulin cork or rubber black tube with a. 

i. amount 1 mg/dispenser. 

To determine the expediency of conducting the protective measures the phytophage 

damage of 7 black currant cultivars is evaluated differing by maturity time. It is 

determined that mid-early cultivar (‘Minay Shmyrev’, ‘Partisanka’) under conditions 

of Belarus are damaged by currant bud moth much stronger than the mid and mid-late 

cultivars. The least pest-damaged cultivar is a mid-late ‘Zolushka’. 

Gauta 2009 06 30 


266

References 

1. Hricovsky I., Spanik F., Repa S. 1994. Analyticka metoda prognoz nastupu fenofag 

cervenej ribezle. Acta fytotechnology, 49: 71–75. 

2. Jonaitis V. 1994. Some aspects of long-term dynamics of phonological situation 

of the various biological systems functioning in different ecosystems. Acta entomologica 

Lituanica, 12: 64–72. 

3. Kajfez-Bogataj L. Bergant K. 1998. Prediction of blossoming of apple tree 

(Malus domestica Borkh.) in phenological models. Zb. Biotehn. Fak. Univ. v 

Ljubljani. Kmetijstvo. Ljubljana. Letn, 71: 83–89. 

4. Koltun N. E., Jarcakovskaja S. I., Supranovich R. V. 2003. Prognosa fenologiczna 

podstawa integrowanej ochrony roslin. Progress in Plant Protection (Postepy 

w Ochronie Roslin). Poznan, 43 (1): 192–199. 

5. Labanowska B. 2003. Krzywik porzeczkowiaczek przypomina o sobie. Haslo 

ogronicze, 2.www.ho.haslo.pl/index/phprok=2003& numer=02-22k. 

6. Morgan D., Solomon M. G. 1993. PEST-MAN: a forecasting system for apple 

and pear pests. Bull. OFPP, 23 (4): 601–605. 

7. Ono S., Konno T. 1999. Estimation of flowering date and temperature characteristics 

of fruit trees by DTS method. JARQ, 33 (2): 105–108. 

8. Zar H. J. 1996. Biostatistical analysis. Prentice-Hall Int. London. 

9. Алехин В. Т., Ермаков А., Черкашин В. И. 1988. Контроль фитосанитарного 

состояния садов и виноградников. Защита и карантин растений, 2: 54–57. 

10. Бородий, С. А., Зубков А. Ф. 2001. Имитационно-статистическое моделирование 

биоценотических процессов в агроэкосистемах. Санкт-Петербург. 

11. Вандова М. 2000. Прогнозиране на цъфтежа при някои овощни видове по 

средната месечна температура. Зумеделие плюс, 5: 11–12. 

12. Грин Н., Стаут У., Тейлор Д. 1996. Количественная экология. В кн.: 

Биология, 2: 127–150. 

13. Крикунова Н. И., Супранович Р. В., Ярчаковская С. И. 2007. Вредители 

и болезни плодово-ягодных, овощных культур и картофеля. Минск: 

Белорусская наука. 

14. Полуэктов Р. А. 2001. Полевой опыт и динамические модели продуктивного 

процесса. В кн. Современные проблемы опытного дела, 1: 29–35. 

15. Савздарг Э. Э. 1960. Вредители ягодных культур. Москва: Госиздат с. х. 

лит. 

16. Уланова Е. С., Забелин В. Н. 1990. Методы корреляционного и 

регрессионного анализа в агрометеорологии. Ленинград. 

17. Ярчаковская С. И. 2000. Вредители смородины и крыжовника. Ахова 

раслiн, 1: 20–21. 

267


Incurvaria capitella Cl. kontroliavimo laiko ir tikslingumo optimizavimas 

S. Yarchakovskaya, N. Kaltun 

Santrauka 

Tyrimai atlikti Minsko regiono juodųjų serbentų plantacijose 1994–2008 metais. Tyrimų 

objektas buvo Baltarusijoje augančios juodųjų serbentų veislės ir serbentinės kandys (Lampronia 

(Incurvaria) capitella Cl.). Tyrimų tikslas – remiantis fenologinėmis kenkėjų vystymosi 

prognozėmis, sukurti serbentinių kandžių kontroliavimo sistemą juodųjų serbentų plantacijose. 

Atsižvelgiant į skirtingą serbentų veislių pažeidimą, buvo panaudotas fitofagų pirminis 

sintetinis lytinis feromonas. Sukurtas fenologinių serbentinių kandžių vystymosi prognozių 

algoritmas. Jis suteikė galimybę iš anksto nustatyti optimalų žiemojimo vietas paliekančių 

vikšrų registravimo laiką, drugių skraidymo dinamiką bei imtis viso komplekso cheminių ir 

agrotechninių priemonių. Ištirtas pirminių sintetinių fitofago lytinių feromonų patrauklumas. 

Nustatyta, kad kaip sintetinio lytinio feromono dalytuvą geriausia naudoti insulino kamštį arba 

juodą guminę žarnelę su 1 mg/dalytuve aktyviosios medžiagos kiekiu. Nustatyta, kad Baltarusijos 

sąlygomis vidutiniškai ankstyvos veislės (‘Minay Shmyrev’, ‘Partisanka’) serbentinių kandžių 

pažeidžiamos daug labiau negu vidutinės arba vidutiniškai vėlyvos. Mažiausiai kenkėjų pažeista 

veislė buvo vidutiniškai vėlyva ‘Zolushka’. 

Reikšminiai žodžiai: feromonas, Incurvaria capitella, juodieji serbentai, kontroliavimas, 

patrauklumas, raidos prognozės, veislių pažeidimas. 

268




Biological activity of plant extracts and their 

application as ecologically harmless biopesticide 

Ivars Zarins 1 , Maris Daugavietis 2 , Julija Halimona 1 

1 

Institute of Biology of University of Latvia, Miera street 3, Salaspils, LV-2169 

Latvia, e-mail iz@email.lubi.edu.lv 

2 

Latvian State Forest Research institute “Silava”, Riga Street 111, Salaspils, 

LV-2169 Latvia, e-mail maris.daugavietis@silava.lv 

The use of plant extracts in the management of plant diseases is gaining importance. 

Different phytopreparations, which were made on the basis of extractive substances originated 

from natural and cultivated plant extracts, are described in this study. Collection of 

phytopreparations consisted of 9 new ones with insecticidal and 8 – with fungicidal properties. 

Phytopreparations were prepared in different forms: liquid, semi-fluid and paste. 

Phytopreparations were tested to be ecologically harmless. 

The fungicidal and insecticidal properties of the phytopreparations were detected under 

laboratory conditions, in the experimental fields, greenhouses and under conditions of peasant 

farm. An efficiency of the tested preparations was found to be 55–81 % in experimental 

fields. 

Besides, an insecticidal activity of “Fitoekols-IF” (liquid and paste form) was tested on 

nine sucking insect species wintering on the bark of fruit trees trunk and basal branches. This 

activity reached 40–78 %. 

New phytofungicides were used against distribution of the pathogenic fungus infections 

on vegetable culture in greenhouses; their efficiency was in the ranges of 65–88 % and 

60–80 % under laboratory and field conditions, respectively. 

Fruits, vegetables and planting material may be successfully protected during their 

long-time storage in storehouses against pathogenic fungus infections with selected phytopreparations, 

e. g. “Fitoekols-IF”, “Fitosativum”, and “Fitocapsicum”. Protection efficiency 

is 58–80 %. 

Key words: bioinsecticide, ecology, phytofungicide, phytoinsecticide, plant extracts, 

plant protection. 

Introduction. Nowadays there still are different chemical pesticides applied in 

plant protection – insecticides, fungicides, herbicides, which provide the fastest and 

highest effect, but at the same time bring on essential ecological damage: pollute soil 

and water sources, destroy entomological communities, cause pathologic changes 

in bird and many warm-blooded animal populations. Wide and persistent usage of 

chemical pesticides is dangerous to human health and causes unhealthy environment 

269

to human work. Especially negative consequences it may cause in fruit and vegetable 

growing, because these products are mostly consumed fresh. Intensive usage of 

chemicals is not suitable in Latvian climatic zone in particular where low intensity of 

sun prolongs distribution of chemical substances in ecosystems. 

To protect different cultural plants from harmful activity of pests and diseases, 

more attention should be paid to developing and establishment of environment friendly 

regulation actions. The quality of products is determined mainly by harmless growing 

techniques applied, which conforms to standards of “ecologically clean” and harmless 

to consumers production. 

Literature studies and our experience suggest that phytopreparations are one of 

the most perspective biological plant protection aids. Their impact is effective enough, 

extraction is not so complicated and time-consuming, they are harmless to environment 

and people, decompose fast in agrocenosis, their usage usually pays off. Basic principle 

of receiving a preparation is adding plant extractive components, which are received 

from plant species immune to pests or diseases. Usually donor plants for receiving 

extract substances are genetically different species, which are widespread in nature or 

cultivated artificially. There is certain success reached in many countries in development 

and practical establishment of new, ecologically safe plant protection aids. 

There are references in literature about fungicidal and insecticidal efficiency of 

different plants, including conifers, and their perceptiveness for production of biological 

preparations (Micales et al., 1994; Biever, 2003; Mares et al., 2004). 

For the last several years Latvian State Forest Research Institute “Silava” and 

LU Institute of Biology have been working at inventing new phytopreparations; besides, 

LFI “Silava” and biological production unit A/S “Biolat” in Piltene, Ventspils 

district, where experimental materials are obtained, improved preparation production 

technology. 

With financial support of Latvian Ministry of Science and Education we produced 

new phytopreparations (“Fitoekols-IF”, “Fitorodents”, “Bembijs”, “Ausma”, “Eko- 

Fit”, etc.), which contain conifer extracts (Daugavietis, 2000 a; Daugavietis, 2000 b; 

Daugavietis, 2001; Daugavietis et al., 2002; Zariņš, Daugavietis, 2002; Daugavietis 

et al., 2005; Зариньш, 2002). 

Our researches suggest, that there are enough wild plant resources and plenty of 

possibilities to grow technical culture of raw material for receiving plant extractive 

substances. By extending this work to make it comply with EU directive requirements, 

it is possible to fully ensure “biological farming” with plant kingdom phytopreparations 

and also to develop the production of preparations for export purposes. 

Object, methods and conditions. Nine phytoinsecticides against sucking pest 

insects were created and tested, which include various extractive substances of wild 

and artificially cultivated plants: “Fitoekols-IF” – (improved form) on pine (Pinus 

sylvestris) and spruce (Picea abies) extracts; “Fitostruts” – greater celandine (Chelidonium 

majus) extracts; “Fitoguns” – on different Ranunculaceae family (Ranunculus 

auricomus, R. polyanthemus, R. scleratus) plant extracts; “Fitoklingerium” – royal 

marigold (Calendula officinalis) extracts; “Fitosamts” – French marigold (Tagetes 

padula, T. erecta) extracts; “Fitopelargonium” – geranium (Pelargonium sp.) 

270

leaf extract; “Fitosinepium” – white mustard (Sinapis alba) plant and seed extract; 

“Fitorusticanum” – wild horse radish (Armoracia rusticana) extract; “Fitotabacum” 

– tobacco (Nicotiana tabacum, N. rustica) extracts. 

Composition of insecticidal preparations: plant extract (in connection with extract 

agent), “Progress” (alkaline product) and sticky substance, in relation 7 : 2 : 1. 

Phytoinsecticides may contain other components – emulgators, cohesive substance, 

etc. Preparations pH is 6.5–7.5. 

The fungicidal properties of preparations were tested under laboratory conditions, 

in greenhouses, experimental fields and under conditions of peasant farm. Activity 

of phytopreparations for limiting numerical composition of pest insect populations 

was determined on following vegetable plants: onion, carrot, cabbage, and pea. 

Prophylactic treatment of plants was begun on 7 th –10 th day after sprouting of seedlings 

and repeated after every 10–14 days, thus managing 3–4 treatments during vegetation 

season. Working concentration of preparations was 1.0–1.5 %. For treating 1 m 2 of 

area 0.5–0.7 l of working suspension is required. For more precise determination of the 

number of insects on model plants and prevention of their migration in agrobiocenosis, 

they are isolated with nylon net. If necessary, supplementary insects are added on 

experimental plants, avoiding exceeding of pest average density on plant. Counting of 

insect numerical composition on both treated and control plants was started on 3 rd –5 th 

day after applying the preparation, and continued throughout vegetation season. 

Many experiments were carried out in fruit-tree gardens. The goal was to find 

out whether these preparations can be used for limiting the population size of sucking 

insects, which overwinter on tree bark or main branches. Liquid forms of preparations 

are brought up on tree by spraying, paste form – with brush. For treatment of 

one 3–5 years-old tree it is necessary approximately 0.4–0.8 l of working suspension 

(depending on the height of tree and the number of main branches). Treatment was 

carried out during autumn period just after leaves fall off. 

The effectiveness of phytopreparations is appraised in spring – April and May – 

visually estimating species density of each pest on the branches and trunk of model 

plants, applying perforated insect-perceptive belts and other methods. In experimental 

variants received data were compared with control results of not treated plants. 

Investigations were carried out in 2000–2008. 

Different sucking pests were examined – aphids, thrips, white flies and spider 

mites in greenhouse (Table 1), in vegetable fields (Table 2, 3) and in fruit tree gardens 

(Table 4). 

Eight phytofungicides were invented to limit infections, caused by phytopathogenic 

fungi on vegetable cultures both in greenhouses and in field, as well as in 

locations of fruit and vegetable crop, decorative cultures and plant material. Active 

component are extracts of various wild and artificially cultivated plants in combination 

with additives – matrixes. 

Plant extracts used for production of phytofungicides were as follows: “Fitoekols- 

IF” – pine (Pinus sylvestris) and spruce (Picea abies) greens extract; “Fitosativum” – 

garlic (Allium sativum) extract; “Fitocapsicum” – chili pepper (Capsicum annuum) 

extract; “Fitokrisanthemium” – chrysanthemum (Chrisanthemum sp.) leaf extract; 

271

“Fitoarmoracium” – wild horse radish (Armoracia rusticana) root and leaf extract; 

“Fitotabacum” – tobacco (Nicotiana tabacum, N. rustica) extracts; “Fitopelargonium” – 

geranium (Pelargonium sp.) leaf extract and “Fitosinepium” – white mustard (Sinapis 

alba) plant and seed extract. 

Fungicidal phytopreparations are produced in different forms – concentrate, liquid, 

paste and dry form. pH of preparations is 8.8–9.0. 

At the same time studies were carried out with already approbated fungicides 

“Bordo”, “Mycostop” and “Timorex”. 

Phytofungicides activity in greenhouses was tested on three vegetable cultures: 

tomatoes, cucumbers and sweet peppers. Five most common infections caused by 

phytopathogenic fungi were chosen: downy mildew Pseudoperonospora cubensis, 

grey mold Botrytis cinerea, leaf mould Cladosporium sp., fusarium wilt Fusarium sp., 

dry leaf spot Macrosporium sp. 

To determine fungicidal preparation effectiveness in vegetable fields, infections 

caused by different phytopathogenic fungi were chosen. 

Working concentration of phytopreparations was 1.0–1.5 %; for treatment of 1 m 2 

area 0.8–1.5 l of working suspension was used. Development stage and intensity of 

fungal infection in test variants was monitored regularly with 4–6 days interval comparing 

received data with control results. Stage of limiting fungal infection development 

was expressed in percents, comparing with plants, which did not undergo treatment. 

On the basis of phytopreparation “Fitoekols-IF” improved preparative form was 

developed – paste, which consists of extractive substance (mixed with extractive 

agent), alkaline product (“Progress”), sticky substance (“PVA-M”), lime or chalk 

powder, CuSO 4 

, KMnO 4 

and emulgator “Tween-7”. Preparation was applied against 

tree cancer – infection, which is caused by phytopatogenic fungus Neustria galligena, 

as well as for prevention of fungal spore spreading in agroecosystem. The effectiveness 

of phytofungicide in fruit gardens is appraised visually estimating the percentage of 

tree cancer comparing to untreated trees. 

Opportunity of applying phytofungicide was clarified to protect fruits and vegetables, 

multiplication material of decorative cultures (flower bulbs, tubers, etc.) from 

fungi caused infections during long-time storage (+5°–+10 °C). Preservation material 

should be treated prophylactically before storage. Seven phytofungicides were tested 

and their protective effect was compared with universal fungicide “Bordo”. Control 

contained material that was not treated with phytopesticides. 

Results. The best results limiting population density of sucking insects on vegetable 

cultures in greenhouses showed phytopreparation “Fitoekols-IF”: technical 

effectiveness on tomatoes and cucumbers was 65–90 %; limiting whitefly population 

size usually it is 95 %. Phytopreparations “Fitostruts”, “Fitosamts” and “Fitosinepium” 

showed a little lower insecticidal activity – 60–91 %. Effectiveness of applying 

preparations “Fitoguns”, “Fitoklingerium” and “Fitopelargonium” on aphids, 

thrips and spider mites did not exceed 65–70 % (Table 1). 

272

Table 1. Insecticidal activity of phytopreparations in greenhouses 

1 lentelė. Insekticidinis fitopreparatų poveikis šiltnamiuose 

Phytopreparations 

Fitopreparatai 

Aphis 

frangulae 

aphids 

amarai 

Myzus 

persicae 

Technical efficiency 

Techninis efektyvumas (%) 

thrips whiteflies 

tripsai baltasparniai 

Heliothrips Parthenothrips 

Trialeurodes 

haemorrhoidalis 

vaporario- 

dracaenae rum 

spider mites 

erkės 

Tetranychus 

telarius 

Tomatoes 

Pomidorai 

Fitoekols- -IF 65.8 ± 0.4 91.0 ± 2.2 68.8 ± 2.4 72.2 ± 4.4 95.7 ± 3.2 88.7 ± 3.7 

Fitostruts 70.0 ± 2.3 89.2 ± 3.2 71.3 ± 2.2 75.5 ± 2.5 99.0 ± 5.1 70.6 ± 2.4 

Fitosamts 78.3 ± 3.2 68.0 ± 2.5 70.0 ± 1.3 67.7 ± 2.3 91.0 ± 5.1 66.5 ± 2.8 

Fitoguns 66.2 ± 2.8 70.1 ± 2.4 56.7 ± 3.2 50.5 ± 3.0 71.0 ± 2.4 58.2 ± 1.1 

Fitotabacum 85.5 ± 4.1 80.0 ± 3.2 98.5 ± 2.1 98.0 ± 2.6 98.3 ± 4.0 90.0 ± 3.2 

Fitosinepium 88.3 ± 3.2 75.5 ± 2.3 81.3 ± 2.1 90.3 ± 4.0 94.5 ± 3.2 88.3 ± 3.5 

Fito-klingerium 62.0 ± 2.4 67.3 ± 2.1 61.2 ± 3.0 55.3 ± 1.3 74.7 ± 2.3 57.7 ± 3.1 

Fito-pelargonium 67.3 ± 2.1 79.5 ± 3.1 75.0 ± 3.2 70.8 ± 2.0 78.3 ± 3.2 67.5 ± 2.4 

Cucumbers 

Agurkai 

Fitoekols- -IF 72.2 ± 3.1 90.7 ± 4.2 65.5 ± 2.3 70.0 ± 1.3 88.2 ± 2.5 95.5 ± 4.1 

Fitostruts 82.2 ± 4.2 91.5 ± 4.0 78.2 ± 4.7 71.8 ± 2.4 87.5 ± 3.2 73.8 ± 2.4 

Fitosamts 75.0 ± 2.5 61.2 ± 2.0 - - 82.2 ± 4.2 60.8 ± 1.7 

Fitoguns 60.3 ± 1.8 67.5 ± 2.6 51.5 ± 1.4 47.7 ± 0.2 69.8 ± 3.2 50.8 ± 1.7 

Fitotabacum 69.3 ± 2.4 78.8 ± 2.2 88.2 ± 3.2 90.2 ± 2.1 95.5 ± 2.2 85.5 ± 2.3 

Fitosinepium 81.2 ± 4.1 76.7 ± 3.2 78.6 ± 2.9 88.5 ± 2.2 88.2 ± 2.4 90.0 ± 2.7 

Fito-klingerium 56.5 ± 1.3 65.5 ± 1.5 55.7 ± 2.2 50.0 ± 2.3 77.3 ± 1.3 60.3 ± 1.5 

Fito-pelargonium 76.5 ± 3.1 85.7 ± 2.5 81.2 ± 2.5 66.5 ± 3.2 87.6 ± 2.0 69.8 ± 2.1 

Results in limitation population density of sucking insects on vegetable cultures 

under field conditions in experimental fields and farm were alike previous ones. The 

highest activity showed improved “Fitoekols-IF”, “Fitosinepium”, “Fitostruts” and 

“Fitoklingerium” – their effectiveness fluctuates between 55 % and 81 %. Little lower 

effectiveness was observed after using “Fitoguns” and “Fitopelargonium” – technical 

activity doesn’t exceed 50–70 %, the last preparation works more like repellent 

(Table 2, 3). 

Dry forms of preparations “Fitotabacum”, “Fitorusticum” and “Fitosinepium” are 

the most functional against early and late cabbage fly, onion and carrot fly: technical 

efficiency reaches 78–92 %. Observations report, that these preparations not only cause 

death of pest insects on treated plants, but also significantly limit their development. 

273

Table 2. Insecticidal activity of phytopreparations limiting sucking pest insect 

populations in experimental fields 

2 lentelė. Insekticidinis fitopreparatų poveikis mažinant vabzdžių kenkėjų populiacijas 

eksperimentiniuose laukuose 

Phyto-preparations 


Aeumerus 

strigatus 

onion 

svogūnai 

Delia 

antiqua 

Delia 

brassicae 

cabbage 

kopūstai 

Delia 

floralis 

Fitoekols-IF (op) 67.8 ± 2.4 53.7 ± 2.3 72.3 ± 2.3 68.5 ± 2.0 65.5 ± 3.2 81.2 ± 2.5 70.0 ± 2.4 

Fitostruts 69.0 ± 2.3 57.8 ± 1.5 76.2 ± 3.2 70.0 ± 2.4 68.8 ± 1.9 74.5 ± 2.5 63.3 ± 2.3 

Fitoguns 57.3 ± 4.0 52.0 ± 3.2 67.7 ± 2.2 63.5 ± 2.2 56.3 ± 2.2 60.0 ± 3.1 51.2 ± 2.6 

Fitosamts 68.2 ± 2.5 65.0 ± 2.2 69.8 ± 2.5 64.2 ± 2.8 60.8 ± 1.9 68.2 ± 2.0 57.7 ± 1.6 

op – original preparation / originalus preparatas 

Table 3. Insecticidal activity of phytopreparations limiting sucking pest insect 

populations under field conditions (farm experiment) 

3 lentelė. Insekticidinis fitopreparatų poveikis mažinant vabzdžių kenkėjų populiacijas 

lauko sąlygomis (gamybinis bandymas) 

Phyto-preparations 


Fitoekols-IF (liquid form 

skystas) 

Fitostruts (liquid form 

skystas) 

Fitosinepium (liquid form 

skystas) 

Fitorusticanum (liquid form 

skystas) 

Fitotabacum (dry form 

sausas) 

Technical effectiveness 


carrot 

morkos 

Brevicoryne 

Trioza Psila 

viridula rosae 

brassicae 

Technical effectiveness (%) after treatment three times per 

vegetative period 

Techninis efektyvumas (%) apdorojus tris kartus per vegetaciją 

onion carrot beat bean peas 

svogūnai morkos burokėliai pupelės žirniai 

Acyrthosiphon 

Delia Psila Trioza Pegomyia Aphis 

antiqua rosae viridula hyoscyami fabae 

onobrichis 

42.2 ± 0.5 57.5 ± 1.9 66.7 ± 2.2 42.7 ± 0.9 70.8 ± 3.1 75.5 ± 2.8 

49.8 ± 1.1 48.8 ± 0.8 70.0 ± 3.0 55.3 ± 1.5 67.3 ± 2.4 70.0 ± 2.5 

52.8 ± 1.8 52.2 ± 2.3 62.2 ± 2.0 44.3 ± 0.7 72.2 ± 3.2 80.0 ± 2.3 

57.5 ± 1.2 47.7 ± 0.9 68.8 ± 2.3 49.8 ± 1.6 65.5 ± 2.7 67.7 ± 3.1 

85.5 ± 2.4 82.7 ± 3.1 58.8 ± 2.1 77.4 ± 2.5 55.5 ± 1.6 46.2 ± 0.9 

“Fitoekols-IF” (liquid form) decreases sucking insect population density under 

critical threshold on fruit trees – 40–67%. Little higher results were given after using 

preparation paste – 61–78 % (Table 4). Oscillations of insecticide effectiveness between 

variants can be explained with treatment quality, insect species, and steadiness of preparations 

on treated plant surface and other factors. It should be pointed out, that usage 

of phytopreparations significantly diminishes development of pests in ecosystem. 

274

Table 4. Insecticidal activity of “Fitoekols-IF” phytopreparation (liquid and paste 

forms) limiting sucking pest insect populations on fruit trees during overwintering 

4 lentelė. Insekticidinis fitopreparato “Fitoekols-IF” (skysto ir pastos pavidalo) poveikis 

mažinant vabzdžių kenkėjų populiacijas vaismedžiuose žiemojimo laikotarpiu 

Protective effect compared to control 

Pest insect 

Teigiamas poveikis, palyginus su kontroliniu variantu (%) 

Vabzdžiai kenkėjai 

liquid form 

skystas 

paste 

pasta 

Lepidosaphes ulmi 62.6 ± 2.2 75.8 ± 3.5 

Plesiocoris rugicollis 51.1 ± 2.4 68.3 ± 2.9 

Lygus pratensis 47.3 ± 2.3 64.2 ± 3.3 

Psylla mali 67.0 ± 3.7 78.0 ± 3.5 

Aphis pomi 56.4 ± 2.5 77.5 ± 4.0 

Cydia pomonella 38.0 ± 2.0 57.0 ± 2.8 

Tmetocera ocellana 43.4 ± 2.1 61.2 ± 3.1 

Yponomeuta malinellus 67.5 ± 2.4 67.5 ± 2.4 

Panonychus ulmi 48.0 ± 2.4 63.0 ± 2.8 

Effectiveness of preparations limiting diseases caused by fungi in greenhouses: 

• “Fitoekols-IF”: limitation of mildew on tomatoes – 75–79 %, on cucumbers – 

75–90 %, on peppers (sweet) – 70–85 %; of grey rot on tomatoes – 68–77 %, on cucumbers 

– 65–68 % and peppers – 70–85 %; of leaf mould on tomatoes – 65–71 %, 

on cucumbers – 65–68 %, of fusarium wilt on tomatoes – 66–68 %, of dry leaf spot 

on tomatoes – 67–76 %, on cucumbers – 58–61 %. 

• “Fitosativum” diminishes mildew on tomatoes for 93–98 %, on cucumbers – 

for 88–90 %, on peppers – for 71–78 %; grey rot on tomatoes – for 89–91 %, on 

cucumbers – for 80–89 % and on peppers – for 85–86 %. 

• “Fitokrisanthemium”: limitation of mildew on tomatoes – 69–77 %, on cucumbers 

– 65–68 %, on sweet peppers – 56–61 %; of grey rot on tomatoes – 75–79 %, on 

cucumbers – 68–69 % and peppers – 53–58 %; of leaf mould on tomatoes – 69–76 %, 

on cucumbers – 64–76 %; of fusarium wilt on tomatoes – 69–76 %; of dry leaf spot 

on tomatoes and cucumbers – 77–81 % and 65–69 %. 

• “Fitoarmoracium” showed effectiveness in fighting mildew on tomatoes is 

90–99 %, on cucumbers – 86–88 %, on peppers – 68–77 %; leaf mould on tomatoes – 

87–95 %, on cucumbers – 77–82 % and on peppers – 67–71 %; fusarium wilt on 

tomatoes 80–89 % and dry leaf spot on tomatoes – 87–90 %. 

• “Fitotabacum” protective effect of fighting mildew on tomatoes is 71–88 %, 

on cucumbers – 73–80 %, on peppers – 65–67 %; grey rot on tomatoes – 71–79 %, 

on cucumbers – 71–80 %, on peppers – 57–60 %; leaf mould on tomatoes – 67–77 %, 

on cucumbers – 64–75 %; fusarium wilt on tomatoes– 67–79 % and dry leaf spot on 

tomatoes – 68–77 %. 

• Using “Fitopelargonium” mildew infection was limited on tomatoes – for 

68–77 %, on cucumbers – for 62–76 %, on peppers – for 66–68 %; grey rot infection on 

tomatoes – 67–73 %, on cucumbers – 74–77 %; fusarium wilt on tomatoes– 74–80 % 

and dry leaf spot on tomatoes – 68–73 %. 

275

• “Fitosinepium” activity limits mildew on tomatoes for 90–99 %, on cucumbers 

– 89–100 %; grey rot on tomatoes – for 96–98 %, on cucumbers – for 90–98 %; 

leaf mould on tomatoes – for 96–99 %, on cucumbers – 90–98 %; fusarium wilt on 

tomatoes – for 85–88 % and dry leaf spot on tomatoes – for 80–86 %. 

From the results of studies it can be concluded that many fungicides (“Fitoekols- 

IF”, “Fitosativum”, “Fitoarmoracium” and “Fitosinepium”) show considerable effectiveness 

in limitation of fungi caused infections on vegetable cultures in greenhouses, 

exceeding 80–95 %. At the same time studies were carried out with already approbated 

fungicides “Bordo”, “Mycostop” and “Timorex”, which showed that their effect doesn’t 

differ significantly from that of newly invented phytopreparations. In many cases new 

phytopreparation effect was even stronger for 12–20 %. 

Comparing effect of phytofungicides under field conditions it can be concluded 

that after their repeated usage during vegetative season in the majority of cases the 

development of disease and its further dispersal is completely stopped. The highest 

efficacy showed “Fitoekols-IF”, “Fitosativum” and “Fitocapsicum” (Table 5). Newly 

invented phytopreparation efficacy doesn’t differ significantly from already existent 

fungicides – “Bordo” and “Timorex”. 

Table 5. Fungicidal activity of phytopreparations limiting infections caused by 

fungi on vegetable cultures under field conditions 

5 lentelė. Fungicidinis fitopreparatų poveikis mažinant grybų sukeltas infekcijas daržovėse 

lauko sąlygomis 





Phoma Botrytis Olpidium Phoma Peronospora 

pisi deni 

P. schlei- 

P. brassicae 

lingam cinerea brassicae betae 

Fitoekols-IF 65–70 60–75 75–85 70–80 55–65 75–80 65–75 

Fitotabacum (df / s) 70–75 70–75 60–75 60–65 65–70 65–72 60–70 

Fitosinepium (df / s) 65–73 55–70 55–70 64–75 50–58 60–65 55–65 

Fitosativum (lf / sk) 70–88 85–90 70–82 72–78 62–70 60–73 56–70 

Fitoarmoracium (lf / sk) 70–77 75–80 72–78 63–70 65–78 62–70 55–65 

Fitosinepium (lf / sk) 55–75 62–68 65–75 50–65 57–70 63–68 58–65 

Fitocapsicum (lf / sk) 72–78 65–70 70–78 75–90 62–68 70–75 65–75 

Fitokrisanthemum (lf / sk) 50–65 55–60 65–70 58–65 50–58 53–58 60–75 

Bordo 70–75 65–70 70–85 65–75 60–75 65–75 55–70 

Timorex 55–68 60–75 58–65 68–72 58–65 60–65 50–65 

df – dry form; lf – liquid form / s – sausas; sk – skystas 

Improved form of “Fitoekols-IF” with good results can be applied fighting against 

fruit tree cancer (caused by N. galligena). Efficacy of preparation is 65–75 %. It should 

be pointed out that phytofungicide also for 70–85 % suppresses fern and lichen growing 

on fruit tree trunks. 

Study results suggest that phytofungicide can be applied to protect fruits and 

vegetables, multiplication material of decorative cultures (flower bulbs, tubers, etc.) 

from fungi caused infections during long-time storage (+5 °–+10 °C). 

276

Table 6. Phytofungicidal activity of phytopreparations limiting infections caused 

by fungi during long-time storage of vegetables, fruits and multiplication 

material 

6 lentelė. Fitofungicidinis fitopreparatų poveikis mažinant grybų sukeltas infekcijas ilgo 

daržovių, vaisių ir dauginimo medžiagos laikymo metu 



Botrytis 

allii 



Botrytis cinerea 

grey rot on cabbage 

kekerinis puvinys ant 

kopūstų 

Alternaria 

radicina 

Phoma 

rostrupii 

B. cinerea 

on carrot 

ant morkų 

Fitoekols-IF 

77.3 ± 2.5 75.0 ± 2.5 68.3 ± 2.4 77.8 ± 2.5 80.5 ± 2.7 

(improved form 

patobulintas) 

Fitotabacum (df / s) 60.0 ± 2.3 54.7 ± 3.1 53.3 ± 2.4 60.2 ± 1.9 65.5 ± 2.1 

Fitosinepium (df / s) 64.3 ± 2.7 59.2 ± 2.0 59.8 ± 1.7 65.5 ± 2.1 76.5 ± 2.5 

Fitosativum (lf / sk) 65.8 ± 2.6 71.5 ± 2.4 69.0 ± 3.2 74.8 ± 2.5 78.2 ± 2.8 

Fitocapsicum (lf / sk) 66.5 ± 2.1 60.2 ± 2.3 57.4 ± 1.7 67.8 ± 2.4 77.3 ± 3.0 

Fitoarmoracium (lf / sk) 61.7 ± 2.5 57.3 ± 1.8 55.8 ± 1.7 64.0 ± 2.1 63.2 ± 2.9 

Fitopelargonium (lf / sk) 55.0 ± 0.7 59.2 ± 2.4 58.5 ± 0.8 60.3 ± 2.0 61.2 ± 2.1 

Bordo (original 7.2 ± 2.1 70.3 ± 2.3 65.7 ± 2.0 68.81.6 80.0 ± 2.3 

originalus) 

df – dry form; lf – liquid form / s – sausas; sk – skystas 

Table 6 shows protective effect of seven phytofungicides to prevent rot infection 

development on different vegetables. The best effect (58–80 %) was reached using 

“Fitoekols-IF” (improved form), “Fitosativum” and “Fitocapsicum”. The effectiveness 

of other phytofungicides was lower – 50–70 %. “Bordo” activity didn’t differ significantly 

from that of newly invented preparations. Applying of preparations in liquid 

form is more time-consuming, because material needs drying after its treatment. 

Discussion. The philosophy of plant protection using plant extracts is to transfer 

organic substances from one plant to another and from one ecosystem to another to 

destroy and exchange the properties of habitual culture medium of pests and fungus. 

The transfer of organic substances between genetic distant kindred plants and 

distant ecosystems may be most effective. The method is more effective against pests 

and diseases with narrow habitual medium. 

Without exchange of the habitual culture phytopreparation layer, which covers 

plant leaf surface, makes it difficult for sucking insects to get to plant cell sap. Stickiness 

of preparation fixates pests to leaf surface, reduces their migration and paralyzes 

physiological processes – feeding, breathing, etc. That is why phytoinsecticides can 

be considered as contact-preparations, and at the same time – as antifeedants, which 

suppress insects feeding and multiplying instincts, as well as their dispersal in environment. 

Fungicidal phytopreparations work owing to their specific properties – alkaline 

medium, stickiness, viscosity and others. Plant absorbs active substances and they 

277

join cell sap circulation system. Thus, phytofungicides work as systemic or intoxication 

preparations and at the same time as contact preparations. Fungal material gets 

suppressed by slowing down the development of spores. Laboratory examinations 

suggest that plant photosynthesis, humidity, air balance and other physiological processed 

are not affected significantly (Theander, 1982; Vyrodov et al., 1987; Zariņš 

and Daugavietis, 2002). 

The insecticidal and fungicidal activity of proposed phytopreparations is similar 

to the preparations elaborated in other countries on the basis of different plant extracts 

(Buver, 2003; Mares, Tosi et al., 2004; Micales, Han et al., 1994). 

Conclusions. Nine phytopreparations with insecticidal properties were worked 

out for limitation of pest insect population size on vegetable cultures and fruit trees 

both in field and greenhouses. The effectiveness of phytoinsecticides in greenhouses 

was 65–95 % depending on vegetable culture, preparation form quality of treatment 

and other factors. 

Phytopreparation effectiveness under field conditions was checked on ten pest 

species and it was 55–81 %. 

Activity of phytoinsecticide “Fitoekols-IF” (liquid and semi-liquid form) was 

defined on nine sucking insect species, which overwinter on fruit tree trunk and main 

branches. It was 40–78 %, depending on preparative form. 

Eight preparations were invented with fungicidal properties for limitation of fungi 

caused infections on vegetables and fruit trees. Their effectiveness in greenhouses 

was 65–88 % (checked on 5 fungal infections); under field conditions it was 60–80 % 

(checked on 7 fungal infections). 

Protective effect of vegetables, fruits and multiplication material of decorative 

cultures from fungal infections in storage places for preparations “Fitoekols-IF”, 

“Fitosativum” and “Fitocapsicum” was 58–80 %, for phytofungicides “Fitotabacum”, 

“Fitosinepium”, “Fitoarmoracium” and “Fitopelargonium”, as well as for “Bordo” 

was a little lower – 50–70 %. 

Phytofungicide “Fitoekols-IF” (paste) supplied with different specific components 

can be successfully applied for limitation of tree cancer (approximately for 60–75 

%), as well for protection from solar radiation and growing of ferns and lichens on 

tree bark. 

All the invented preparations as active substances include extracts, which are 

received from wild or artificially cultivated plants and are supplied with different 

matrixes – cohesive substances, sticky substances, emulgators, etc. Phytopreparations 

are ecologically safe, do not leave negative effect on treated plants, their usage pays 

off and they are suitable for prophylactic treatment. 

Gauta 2009 06 


278

References 

1. Biever C. 2003. Herb extracts wrap up lethal food bugs. New Scientist, 

178(2 399): 26–27. 

2. Daugavietis M. 2001. Experience of pine and spruce needle” s extractive using 

for plants protections. In: Proceedings of the International Conference. Estonia, 

Tartu, 11–13. 

3. Daugavietis M. 2000 a. Non-wood, tree biomass – a raw material of coming century. 

In: Proceeding of World Congress “State of knowledge report for IUFRO 

XVI World Congress, 3 rd division”, Malaysia, 117–121. 

4. Daugavietis M. 2000 b. Tree biomass as raw material for active substance. In: 

Proceedings of International Conference “Wood-material for all times”. Poland. 

Rogovo. 93–97. 

5. Daugavietis M., Polis O., Korica A. 2002. Priede –augstvērtīgu bioloģiski aktīvu 

dabas vielu avots. LLU Raksti, 5: 59–67. 

6. Daugavietis, M., Polis, O., Korica, A. 2005. Egles vainaga biomasas izmantošanas 

iespējas. LLU Raksti, 14(309): 72–75. 

7. Mares D., Tosi B., Poli F., Andreotti E., Romagnoli C. 2004. Antifungal activity 

of Tagetes patula extracts on some phytopathogenic fungi: ultrastructural evidence 

on Pythium ultimum. Microbiology Research, 159(3): 295–304. 

8. Micales I. A., Han I. S., Davis I. L., Young R. A. 1994. Chemical composition 

and fungitoxic activities of pine cone extractives. Biodeterior, 4: 317–332. 

9. Theander Olof. 1982. Hydrophilic activities from the needles of Scots pine and 

Norwey spuce. Upsala, Sweden. Journal General Review Papperstidn, 85(9): 

64–68. 

10. Vyrodov V. A., Solodkaja G. F., Nikolaeva M. A., Smirnova L. D. 1987. 

Fungicidal properties of preparations extracted from pine and spruce with isoprophyl 

alcohol. (In Russian, from Ref. Zh., Khim. Abstract, 24–37). 

11. Zariņš I., Daugavietis M. 2002. The “Fitorododents-RF” – new environmentally 

friendly repellent of animals in the garden and agrobiocenoses. In: Environmental 

and Crop Production, 21–27. 

12. Zariņš, I., Daugavietis, M. 2002. The “Fitorododents-RF” – new environmentally 

friendly repellent of animals in the garden and agrobiocenoses. In book: 

“Environmental and Crop Production”, 21–27. 

13. Зариньш И. А. 2002. Эффективность некоторых новых биопрепаратов в 

защите растений. Защита растений, 6: 35–36. 

279


Augalų ekstraktų biologinis efektyvumas ir jų naudojimas kaip ekologiškai 

nekenksmingų biopesticidų 

I. Zarins, M. Daugavietis, J. Halimona 

Santrauka 

Augalų ekstraktų naudojimas, siekiant apsaugoti augalus nuo ligų, darosi vis svarbesnis. 

Šiame straipsnyje aprašyti įvairūs fitopreparatai, pagaminti iš gamtoje augančių ir sukultūrintų 

augalų ekstraktų. Fitopreparatų rinkinys susidėjo iš 9 naujų fitopreparatų su insekticidinėmis 

savybėmis ir 8 – su fungicidinėmis savybėmis. Fitopreparatai buvo paruošti skirtingų formų – 

skysti, pusiau skysti ir pastos pavidalo. Buvo patikrinti, ar yra ekologiškai nekenksmingi. 

Fungicidinės ir insekticidinės fitopreparatų savybės tirtos laboratorijoje, eksperimentiniuose 

laukuose, šiltnamiuose ir ūkininko ūkyje. Eksperimentiniuose laukuose tirtų preparatų 

efektyvumas buvo 55–81 %. 

Be to, insekticidinis “Fitoekols-IF” (skysto ir pastos pavidalo) poveikis buvo išbandytas 

su devynių rūšių vabzdžiais kekėjais ant vaismedžių žievės ir pagrindinių šakų. Šis poveikis 

siekė 40–78 %. 

Nauji fitofungicidai buvo naudoti, siekiant išvengti patogeninės grybinės infekcijos 

plitimo ant daržovių šiltnamiuose. Jų veiksmingumas laboratorijos sąlygomis buvo 65–88 %, 

laukuose – 60–80%. 

Ilgalaikio laikymo saugyklose metu vaisiai, daržovės ir sodinukai gali būti sėkmingai 

apsaugoti nuo patogeninės grybinės infekcijos pasirinktais fitopreparatais, pvz., “Fitoekols-IF”, 

“Fitosativum”, ir “Fitocapsicum”. Apsaugos efektyvumas – 58–80 %. 

Reikšminiai žodžiai: augalų apsauga, augalų ekstraktai, bioinsekticidai, ekologija, 

fitofungicidai, fitoinsekticidai. 

280




Detection and characterization of Cucumber mosaic 

virus isolated from sweet peppers 

Irena ZITIKAITĖ, Marija SAMUITIENĖ 

Institute of Botany, Žaliųjų Ežerų 49, LT-08406, Vilnius, Lithuania, 

e-mail: irena.zitikaite@botanika.lt; marija.samuitiene@botanika.lt 

Cucumber mosaic virus (CMV) causing viral diseases in forage, fruit, ornamental and 

vegetable crops worldwide has been isolated in Lithuania from sweet pepper (Capsicum annuum 

L.) plants exhibiting mottle-mosaic and distortion of leaves and fruits, and plant stunt 

symptoms. The plant material was collected in the private gardens of Vilnius, Kaišiadorys, 

Kėdainiai regions. The identification of CMV has been performed on the basis of determination 

of host range, symptom expression on the test plant species and morphological properties 

of the virus particles by the methods of test plants and transmission electron microscopy, 

and by using of specific oligonucleotide primers in reverse transcription-polymerase chain 

reaction (RT-PCR). In this work the primers designed on the basis of published sequences 

were applied for amplification of CMV RNA fragments in RT-PCRs using experimentally 

CMV infected host plants. The detection of CMV in inoculated test plants was confirmed 

by RT-PCR technique. Analysis of PCR products in acrylamide gel electrophoresis revealed 

amplification of about 540 bp (base pair) fragments which were in agreement with size of the 

fragment expected from the sequence data. 

Key words: Cucumber mosaic virus, isolation, RT-PCR, sweet pepper. 

Introduction. Sweet pepper (Capsicum annuum L.) is now grown worldwide 

under various enviromental and climatic conditions and is the second most important 

crop among solanaceous fruits. Observation showed that the production of this crop 

has been banned with viral infection. Viral diseases are the major limiting factors for 

successful pepper cultivation in the world (Francki, 1979; Fujisawa et al., 1986; Florini, 

Zitter, 1987). Viruses that may occur on peppers include Tobacco mosaic, Cucumber 

mosaic, Potato Y, Tomato spotted wilt, Alfalfa mosaic, Pepper mottle, Pepper 

veinal mottle, Pepper ringspot and others (Šutic et al., 1999; Hiskias et al., 1999; Buzkan 

et al., 2006; Ryu et al., 2009). In order of importance are Cucumber mosaic virus 

(CMV). It as a type species of Cucumoviruses is reported to infect 1287 plants species 

in 518 genera belonging to 100 families (Edwardson, Christie, 1997). It is geographically 

wide spread and has been reported in Europe, Australia, North America. It is one 

of the most important virus disease agent of pepper worldwide. It is transmitted by numerous 

species of aphid in a non-persistent manner (Francki et al., 1979; Kaper, Wa- 


terworth. 1981). CMV is not transmitted through pepper seeds. It also has an extremely 

wide host range and causes fern leaf, stunting of pepper and malformation of fruits. 

Morphologically CMV has rather characteristic about 30 nm polyhedral particles 

with hollow center (Palukaitis et al., 1992). CMV particles contain about 18 % RNA. 

The virions are not stable to freezing. Long-term storage of CMV is most reliable in 

the form of viral RNA, which is highly infectious, and very stable at -20 ° C (Foster, 

Taylor, 1998). Great number of different CMV strains and serogroups has been described 

(Kaper, Waterworth, 1981; Perry et al., 1993). In Lithuania, this virus spread 

on leguminous (Staniulis, 1994), ornamental (Navalinskienė, Samuitienė, 2006), 

cucumber and tomato (Zitikaitė, 2002; Zitikaitė et al., 2006) plants. 

The purpose of this work was to study properties of virus isolates extracted from 

sweet pepper samples and to identify the causal agent. 

Object, methods and conditions. Twelve leaf samples of cultivated sweet 

pepper were collected by visual screening of greenhouses and grown fields under 

plastic on presence of symptoms of viral etiology. Leaves showing disease symptoms 

were collected in plastic bags, kept at 4 °C, when triturated in 0.1 M sodium 

phosphate buffer at pH 7.0–7.1 and rubbed on 600 mesh carborundum dusted leaves 

of test plants for virus propagation. For investigation of virus host range and induced 

symptoms about twenty herbaceous plant species from families of Solanaceae 

Juss., Cucurbitaceae Juss., Aizoaceae Rudolphi, Chenopodiaceae Vent. were tested 

as indicator plants (Table) (Matthews, 1993). Copper grids were floated on drops 

of crude extract of virus infected plants for 1 to 2 minutes, rinsed with bidistilled 

water and subsequently stained with 3 % uranyl acetate. Grids were examined under 

a JEOL-100S transmission electron microscope (EM) (Dijkstra, de Jager, 1998). 

For detection of CMV isolated from sweet pepper plants by RT-PCR technique 

the frozen plant material was used. Nucleic acids of CMV were extracted using the 

small-scale procedure as proposed for extraction of nucleic acids from woody plants 

(Zhang et al., 1998) with slight modifications. Tissue samples of infected test-plants 

were ground in liquid nitrogen and transferred to microtubes. 600 µl 1 × STE buffer 

(0.1 M Na Cl, 0.001 M Tris, 0.001 EDTA, pH 6.9), 80 µl of 10 % SDS and and 800 µl 

of 2 × STE-saturated phenol was added to the powdered tissues. The mixture was 

centrifuged 5 min at 16 000 g. Aqueous phase was removed and transferred to a clean 

microfuge tube. Ethanol to a final concentration of 30 % was added, then ~ 10 mg 

cellulose (whatman CF-11). Cellulose CF-11 was washed by vortexing 3 times with 

1 ml of 1 × STE/30 % ethanol, collecting cellulose by centrifugation between washes 

and discarding supernatants. RNA from cellulose CF-11 was eluted by adding 200 µl 

of 1 × STE buffer, and centrifugation for 5 min. Supernatant was transferred to a clean 

tube. For precipitation of the RNA 40 µl of 3 M sodium acetate and 1 ml of ethanol was 

added. The tube was incubated at -20 °C for 2 h., centrifuged for 10 min at 16 000 g, 

and the pellet was incubated with 80 % of ethanol at -20 °C, and air-dried. 

Primer pair for CMV detection by RT-PCR was used: D, 5‘ – GCG CGA AAC 

AAG CTT CTT ATC – 3‘ (nt 633 to 653) and U, 5‘ – GTA GAC ATC TGT GAC GCG 

A – 3‘ (nt 114 to 132) (de Blass et al., 1994). Pellets of total RNA were resuspended in 

the solution containing 1 % RNAse inhibitor, 0.4 µM primer Reverse and PCR water 


and incubated at 70 °C for 10 min. For the first strand copy DNA synthesis the RNA 

pellet solutions to the mixture containing 5 × Reaction buffer, RNAse inhibitor, dNTP 

mixture and RevertAid TM M-MuLV Reverse Ttranscriptase (MBI Fermentas, Lithuania) 

were added. The first strand cDNA synthesis was carried out at 37 °C for 60 min and 

70 °C for 10 min. DNA amplifications were performed in reaction mixtures containing 

PCR water, 10 mM dNTP mixture, both primers, 10 × PCR buffer with MgCl 2 

and recombinant Taq polymerase (MBI Fermentas) using Eppendorf Mastercycler 

Personal. PCRs were carried out for 40 cycles using the following parameters: 1 min 

at 94 °C (4 min for the first cycle), 2 min at 55 °C and primers extension for 2 min 

(10 min in the final cycle) at 72 °C (Saiki et al., 1988). DNA fragment size standard 

was × 174DNA/BsuRI(HaeIII) digest (MBI Fermentas) (from top to bottom: 1 353, 

1 078, 872, 603, 310, 281, 271, 234, 194, 118, 72 bp). Resulting PCR products were 

analysed by electrophoresis through 5 % polyacrylamide gel, stained with etidium 

bromide (EB), and DNA bands were visualized under UV light. 

Results. Symptoms of naturally affected sweet pepper (C. annuum L.) plants 

vary widely. One of the most common expressions was a severely stunted. Plants 

have light green foliages. In some cases the leaves become narrow and no longer 

expand, while in other cases small necrotic spots with oak leaf patterns develop. 

Leaves were smaller than normal and mild mottled (Fig. 1). Older plants of sweet 

pepper show foliar mottling followed by diffuse chlorosis or no symptoms on foliage 

or fruit. Fruits of some sweet pepper were deformed and reduced in size and form. 

Fig. 1. Sweet pepper leaves affected with CMV 

1 pav. CMV pažeisti saldžiųjų paprikų lapai 

CMV infected practically all of 20 mechanically inoculated test plants in four isolates 

(Table). The pathogen caused vein brightening, mottling and various deformations 

of growing leaves of Nicotiana glutinosa L., N. rustica L. (Fig. 2). Systemic reaction 

in the form of growth disorder, mosaic or mottling and deformation of young leaves of 

infected N. tabacum L. ‘Samsun’ was also noticed. CMV infection developed the most 


conspicuous symptoms on the leaves of Datura stramonium L. – diffusive changeable 

light and dark green areas (Fig. 3). In the leaves of test plant of Chenopodium L. genus 

a local reaction in the form white or chlorotic lesions was revealed (Fig. 4). Under 

the influence of CMV only local necrotic lesions appeared on the leaves of Nicandra 

physalodes (L.) Gaertn (Fig. 5). Virus developed diffusive chlorotic spots, which later 

formed up a clear mosaic picture on the upper leaves of Cucumis sativus L. 

Numerous isometric particles with hollow center were commonly observed in leaf 

dip EM preparations of leaf samples of infected plants. They were about 28–30 nm 

in diameter (Fig. 6). 

Fig. 2 / 2 pav. Nicotiana rustica (S) 

Fig. 3 / 3 pav. Datura stramonium (S) 

Fig. 4 / 4 pav. Chenopodium 

amaranticolor (chlLL) 

Fig. 5 / 5 pav. Nicandra 

physalodes (nLL) 

Table. Test plant reaction to CMV isolated from sweet peppers 

Lentelė. Augalų indikatorių reakcija į CMV, izoliuotą iš saldžiųjų paprikų 

Virus isolates and symptoms 

Test plants 

Izoliatai ir simptomai 

Augalai indikatoriai 

9007 9707 0211 0413 

1 2 3 4 5 

Amaranthus caudatus 

Capsicum annuum ‘Kristal’, 

‘Podarok Moldovy’ 

L: nLL S: 

Mo, Ma; 

S: Mo, Dis 

- 

S: VC, Cr; 

S: M, Mo 

0 

S: VC, Mo 

S: Mo, Dis 

L: nLL 

S: VC, Ru; 

S: VN, Cr 




1 2 3 4 5 

- L: nLL L: nLL 

L: chlLL L: chlLL L: chlLL 

L: nLL 0 

L: nLL 


S: M, Ma S: VC, Ru, Mo S: VC, Mo 

S: M, chlMo S: VC, Cr S: yM 

L: nLL L: nLL L: nLL 

S: Mo, Dis S: Mo, Ru S: Mo, TW 

L: nLL L: nLL 0 

S: Mo S: VC, Dis S: Ru, St 

S: VC, M, Ru S: VC, Mo, Dis S: M, Ru 

S: VC, Cr 0 

S: Mo, Ru 

S: VC, Mo S: M, Mo S: Cr, Dis 

L: difSp; S: Epi S: chlSp S: difSp 

S: chlMo S: difMo S: M, Mo 


S: Mo S: Mo S: VC 

Celosia argentea f. сristata 

Chenopodium amaranticolor 

C. ambrosioides 

C. quinoa 

Cucumis sativus ‘Visconsin’ 

Datura stramonium 

Gomphrena globosa 

Lupinus albus 

Nicandra physalodes 

Nicotiana debneyi 

N. glutinosa 

N. rustica 

N. tabacum ‘Xanthi’ 

Phaseolus vulgaris ‘Bataaf’ 

Pisum sativum ‘Žalsviai’ 

Tetragonia expansa 

Vicia faba ‘Aušra’ 

L: nLL 

L: chlLL 

L: nLL 

L: chlLL 

S: difMo 

S: M, Mo 

L: nLL 

- 

L: nLL 

S:VC, difMo 

S: M, Mo 

S: difMo 

S: VN, Mo 

S: VC, Mo 

S: chlMo 

L: chlLL 

S: Mo 

Abbreviations: L – local reaction, S – systemic reaction, nLL – necrotic local lesions, chlLL – 

chlorotic local lesions, whLL – white local lesions, M – mosaic, Mo – mottling, Ma – malformation, 

VC – vein clearing, VN – vein necrosis, Dis – distortion, dif – diffuse, Cr – crincling, 

Ru – rugosity, Epi – epinasty, TW – top wilting, y – yellow, Sp – spotting, St – stunt, 0 – no 

symptoms, ─ - not tested. 

Santrumpos: L – vietinė reakcija, S – sisteminė reakcija, nLL – nekrotinės vietinės žaizdos, chlLL – chlorotinės 

vietinės žaizdos, whLL – balkšvos vietinės žaizdos, M – mozaika, Mo – margumas, Ma – neišsivystęs, 

VC – gyslų išryškėjimas, VN – gyslų nekrozė, Dis – išsikraipymas, dif – išskydęs, Cr – garbanė, 

Ru – raukšlėtumas, Epi – išaugos, TW – viršūnės vytimas, y – geltonas, Sp – dėmėtumas, St – žemaūgė, 

0 – be simptomų, - – netestuota. 

Fig. 6. CMV particles 

6 pav. CMV dalelės 


Fig. 7. DNA products amplified in PCRs from pepper infected by CMV: 

1 and 11 – DNA Ladder; 2 – healthy plant; 3 and 5 – CMV infected test plants; 

4 – control 

7 pav. PGR amplifikuoti DNR produktai iš CMV pažeistų paprikų: 1 ir 11 – DNR markeris; 

2 – sveikas augalas; 3 ir 5 – CMV užkrėsti augalai; 4 – kontrolė 

RT-PCR using specific primer pair for CMV detection primed amplification of 

about 540 bp DNA sequences from two CMV samples from refrigerated symptomatic 

D. stramonium and N. glutinosa plant tissues (Fig. 7). A sample of non-infected D. stramonium 

plant did not yield visible specific DNA band. Amplification was not observed 

in sample with negative control (PCR buffer and PCR water), too. The molecular 

investigation confirmed that sweet pepper plants have been infected by the CMV. 

Discussion. The experimental host range and specific symptoms on all test plants 

indicated that the virus isolated from sweet pepper crop most closely correspond with 

the CMV (Francki et al., 1979). Based on particles size and morphology, the virus was 

considered to be a member of the Cucumovirus genus (Kaper, Waterworth, 1981). RT- 

PCR data confirmed identification obtained by investigation of the host range, symptomotology 

and virus morphology. RT-PCR product size of CMV isolates from sweet 

pepper showed identity with CMV isolates, identified in other plants in Lithuania. 

CMV is among the most economically damaging pathogens in cucurbits and other 

vegetable crops. Besides cucumber and pepper plants, CMV naturally affects tomato, 

parsley, celery, potato, pea, bean, lupine, clover, fruit and ornamental plants. Several 

aphids readily transmit CMV non-persistently in nature. Among more than 60 aphid 

species vectors, the most efficient are Myzus persicae Sulz., Aphis gossypii Glov., 

A. craccivora Koch. and A. fabae Scop. The large population of aphis vectors is one 

reason for the widespread nature of CMV. CMV spreads through the sap of infected 

plants by leaf contact, through the seeds of 19 plant species and dodder (Francki et al., 

1979; Brunt et al., 1996). 


Conclusions. The virus disease agent detected in sweet peppers in Lithuania has 

been identified as a CMV by using of different virological methods. Sweet pepper is 

a new natural host of CMV in Lithuania. 

Gauta 2009 06 30 


References 

1. Brunt A. A., Crabtree K., Dallwitz M. J., Gibbs A. J., Watson L. 1996. Viruses of 

plants. Descriptions and Lists from VIDE Database. Cambridge. 

2. Buzkan N., Denir M., Oztekin V., Mart C., Caglar B. K., Yilmaz M. A. 2006. 

Evaluation of the status of capsicum viruses in the main growing regions of 

Turke. Bulletin OEPP, 36(1): 15–19. 

3. De Blas C., Borja M. I., Saiz M., Romero J. 1994. Broad spectrum detection of 

cucumber mosaic virus (CMV) using the polymerase chain reaction. Journal of 

Phytopathology, 141: 323–329. 

4. Dijkstra J., de Jager C. P. 1998. Practical Plant Virology. Protocols and Exercises. 

Springer. 

5. Edwardson J. R., Christie R. G. 1987. Viruses infecting forage legumes. 

Cucumoviruses. Gainesville, University of Florida Press. 

6. Edwardson J. R., Christie R. G. 1997. Viruses infecting peppers and other 

Solanaceous crops. Gainesville. University of Florida Press. 

7. Florini D. A., Zitter T. A. 1987. Cucumber mosaic virus (CMV) in peppers 

(Capsicum annuum L.) in New York and associeted yield losses. Phytopathology, 

77: 652. 

8. Forster G. D., Taylor S. C. 1998. Plant Virology Protocols from virus isolation 

to transgenic resistance. Methods in Molecular Biology. Totowa, New Jersey, 

81: 571. 

9. Francki R. I. B., Mossop D. W., Hatta T. 1979. Cucumber mosaic virus. CMI/ 

AAB Descriptions of plant viruses. 

10. Fujisawa I., Hanada T., Saharan A. 1986. Virus diseases occuring on some vegetable 

crops in Spain. Journal of Phytopathology, 125: 67–76. 

11. Hiskias Y., Lesemann D. E., Vetten H. J. 1999. Occurrence, distribution and relative 

importance of viruses infecting hot pepper and tomato in the major growing 

areas of Ethiopia. Journal of Phytopathology, 147: 5–11. 

12. Kaper J. M., Waterworth H. E. 1981. Cucumoviruses. In: Kurstak E. (ed.) 

Handbook of plants virus infections. Elsevier/North Holland Biomedical Press, 

257–332. 

13. Matthews R. E. E. (ed.) 1993. Diagnosis of plant virus diseases. Boca Raton Ann 

Arbor London Tokyo, CRC Press. 

14. Navalinskienė M., Samuitienė M. 2006. Dekoratyvinių augalų virusinės ligos ir 

jų sukėlėjai Lietuvoje. Lututė, Kaunas. 


15. Palukaitis P., Roossinck M. J., Diecgen R. G., Francki R. I. B. 1992. Cucumber 

mosaic virus. Advances in Virus Research, 41: 280–348. 

16. Perry K. L., Habili N., Dietzgen R. G. 1993. A varied population of cucumber 

mosaic virus from peppers. Plant Pathology, 42: 806–810. 

17. Ryu J. G., Ko S. J., Lee Y. H., Kim M. K., Kim K. H., Kim H. T., Choi H. S. 

2009. Incidence and distribution of virus diseases on paprika (Capsicum annuum 

var. grossum) in Jeonnam province of Korea. Plant Pathology Journal, 25(1): 

95–98. 

18. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., 

Mullis K. B., Erlich H. A. 1988. Primer-directed enzymatic amplification of 

DNA with a thermostabile DNA polymerase. Science. 239: 487–491. 

19. Staniulis J. 1994. Ankštinių augalų virusinių ir geltos tipo ligų sukėlėjai Lietuvoje 

(Gamtos mokslų habilitacinis darbas). Vilnius. 

20. Šutic D. D., Ford R. E., Tošic M. T. 1999. Handbook of Plant Virus diseases. 

New York. CRC Press. 

21. Zhang J. P., Uyemoto J. K., Kirkpatrick B. C. 1998. A small-scale procedure for 

extracting nucleic acids from woody plants infected with various phytopathogens 

for PCR assay. Journal of Virological Methods, 71: 45–50. 

22. Zitikaitė I. 2002. Viroses of cucumber plants and identification of their agents. 

Biologija, 2: 42–46. 

23. Zitikaitė I., Survilienė E., Būtaitė G. 2006. Pomidorų (Lycopersicon esculentum 

Mill.) virusų diagnostika elektronomikroskopiniu ir molekuliniu metodais. 

Sodininkystė ir daržininkystė, 25(4): 230–239. 


Iš saldžiųjų paprikų izoliuoto agurkų mozaikos viruso 

nustatymas ir charakterizavimas 

I. Zitikaitė, M. Samuitienė 

Santrauka 

Pasaulyje pašarinius, vaisinius-uoginius, dekoratyvinius ir daržovinius augalus pažeidžiantis 

agurkų mozaikos virusas (Cucumber mosaic virus, CMV) buvo izoliuotas iš saldžiųjų 

paprikų (Capsicum annuum L.) Lietuvoje. Augalai buvo žemaūgiai, margai mozaikiškais ir 

deformuotais lapais bei vaisiais. Tyrimams medžiaga surinkta privačiuose daržuose Vilniaus, 

Kaišiadorių, Kėdainių rajonuose. Virusas identifikuotas pagal simptomų pasireiškimą augaluose 

indikatoriuose, pažeidžiamų augalų spektrą ir virionų morfologiją, panaudojant augalų indikatorių, 

peršviečiamosios elektroninės mikroskopijos bei atvirkštinės transkriptazės-polimerazės 

grandininės reakcijos (AT-PGR) testus. Nustatytas platus viruso pažeidžiamų augalų spektras. 

Labai specifinė simptomatika ir virionų morfologija yra būdingi CMV. Šiame darbe pritaikius 

publikuotų nukleotidinių sekų pagrindu parinktą specifinių oligonukleotidų porą, elektroforezės 

akrilamidiniame gelyje išryškėję kopijinės DNR PGR produktų fragmentai (~ 540 bp 

dydžio) atitiko panaudotus pradmenis ir patvirtino CMV saldžiosiose paprikose infekciją. 

Reikšminiai žodžiai: agurkų mozaikos virusas, AT-PGR, izoliavimas, saldžioji paprika. 


ATMINTINĖ AUTORIAMS, RAŠANTIEMS 

Į MOKSLO DARBUS „SODININKYSTĖ IR DARŽININKYSTĖ“ 

Straipsnio rankraščio pateikimo–priėmimo procedūra 

Straipsnius redakcijai gali pateikti bet kurie Lietuvos ar užsienio šalies mokslo 

darbuotojai bei asmenys, dirbantys mokslinį darbą. Ne mokslo darbuotojo straipsnis 

turi būti parašytas kartu su mokslo darbuotoju. 

Rankraštis redakcijai siunčiamas paštu dviem egzemplioriais, atspausdintas 

kompiuteriu popieriuje, laikantis toliau tekste nurodytų reikalavimų. Pateiktas 

straipsnio rankraštis užregistruojamas ir perduodamas redaktorių kolegijos nariui, 

kuruojančiam šią sritį. Jis įvertina, ar rankraščio turinys ir forma atitinka 

svarbiausius periodiniams straipsniams keliamus reikalavimus. Rankraščiai, kurie 

buvo atmesti pirmojo vertinimo metu, su aiškinamuoju raštu grąžinami autoriui. 

Jeigu straipsnio tinkamumas nekelia abejonių, redaktorių kolegijos narys skiria 

du recenzentus. 

Pataisytą rankraštį autorius turi atsiųsti el. paštu arba paštu redakcijai kartu su 

elektronine laikmena (diskeliu arba kompaktiniu disku) per dešimt dienų. 

Struktūra ir apimtis 

Rankraščio reikalavimai 

Rankraščio forma turi atitikti periodiniams moksliniams straipsniams keliamus 

reikalavimus. Teksto ir jo sudedamųjų dalių seka tokia: 

– Straipsnio pavadinimas (ne daugiau kaip 10 žodžių) 

Pavadinimas rašomas mažosiomis raidėmis paryškintu šriftu (Augalų adaptacijos 

prie šalčio molekulinio mechanizmo aspektai). 

– Autoriaus vardas, pavardė 

Vardas, pavardė – mažosiomis raidėmis paryškintu šriftu (Vyktor Sharma). 

Jeigu yra keli autoriai, rašoma mažėjančia jų autorystės indėlio tvarka. 

– Institucija, adresas, elektroninis paštas 

Rašomi mažosiomis raidėmis kursyvu (Italic) (Lietuvos sodininkystės ir 

daržininkystės institutas). 

Pagrindinis tekstas 

– Santrauka (iki 1 400 rašybos ženklų, arba 250 žodžių) 

Labai glaustai pateikiami tikslai, sąlygos, svarbiausi rezultatai. 

– Reikšminiai žodžiai (ne daugiau kaip 10) 

– Įvadas 

Trumpai išdėstoma nagrinėjama problema, ankstesni kitų panašių tyrimų 

rezultatai, darbo reikalingumas, originalumas. Nurodomas darbo tikslas. 

– Tyrimo objektas, metodai ir sąlygos 


– Rezultatai 

Trumpai išdėstomi tyrimų metu surinkti duomenys, dokumentai (lentelės, 

grafikai). 

– Aptarimas 

Aptariami, bet ne kartojami „Rezultatų“ skyrelyje pateikti duomenys, palyginami 

su kitų autorių duomenimis, aiškinamos tirtų reiškinių priežastys, keliamos 

naujos idėjos, hipotezės. 

– Išvados 

– Padėka (neprivaloma) 

– Literatūra 

Rekomenduojama į sąrašą įtraukti ne mažiau kaip 10 naujausių literatūros 

šaltinių rašoma tema. 

– Santrauka lietuvių ir anglų kalbomis (600–1 400 sp. ženklų) 

Straipsnių, kurie parašyti remiantis netradiciniais bandymų duomenimis ir jų 

rezultatais, struktūrinės teksto dalys gali būti ir kitokios. 

Straipsnis turi būti ne daugiau kaip 10 puslapių apimties kompiuteriu rinkto 

teksto, įskaitant lenteles ir paveikslus (didesnės apimties straipsniai derinami su 

redaktorių kolegijos pirmininku). 

Teksto parengimas 

Straipsnis rašomas lietuvių ir anglų kalbomis ir spausdinamas Microsoft 

WORD teksto redaktoriumi Windows 2000, XP ar VISTA operacinėse sistemose. 

Tekstas rašomas Times New Roman 12 dydžio šriftu, A4 formato 

(210 × 297 mm) lape, atstumas tarp rankraščio eilučių – 1 (single), išlyginamas iš 

abiejų pusių. Paraščių plotis: viršuje – 2 cm, apačioje – 2 cm, dešinėje –1,5 cm, 

kairėje – 3 cm. Straipsnis pateikiamas elektronine laikmena. 

Paryškintai (Bold) rašoma: straipsnio pavadinimas visomis kalbomis, antraštės 

bei svarbiausi struktūros elementai (įvadas, tyrimo objektas, metodai ir sąlygos, 

rezultatai, aptarimas, išvados, padėka, literatūra, santrauka). Kursyvu (Italic) 

rašomi lotyniškieji augalų rūšių, genčių, ligų, kenkėjų, mikroorganizmų ir kitų 

biologinių objektų pavadinimai. Augalų veislių pavadinimai rašomi viengubose 

kabutėse (pvz., ‘Auksis’). 

Cituojamas šaltinis tekste nurodomas lenktiniuose skliaustuose (autoriaus 

pavardė, metai). 

Lentelės 

Lentelėse neturi būti kartojama paveiksluose ar kitose iliustracijose pateikta 

informacija. 

Lentelių tekstas rašomas lietuvių ir anglų kalbomis Times New Roman 

12 dydžio šriftu, jeigu parašyti tekstai talpinami vienoje eilutėje, tarp jų dedamas 

ženklas /. Lentelės teksto dalys vertikaliomis ir horizontaliomis linijomis neatskiriamos. 

Horizontaliomis linijomis atskiriamos tik lentelės metrikos dalys ir lentelės 

pabaiga. Lentelės padėtis puslapyje tik vertikali (Portrait). 

290

Bandymų veiksnių gradacijos lentelėse neturi būti žymimos skaičiais, sudėtingomis 

santrumpomis, o pateikiamos visa arba suprantamai sutrumpinta aprašo 

forma. Pateikiama santrumpa turi būti paaiškinama pirmosios lentelės (paveikslo) 

apraše. 

Statistiniai duomenys, skaičiai ir skaitmenys 

Pageidautina detaliai aprašyti taikytus tyrimų metodus ir nurodyti jų originalius 

šaltinius. Labai svarbi informacija apie lauko, vegetacinių ir kt. bandymų 

išdėstymo schemą ir jos pasirinkimo motyvus. Lentelėse ir paveiksluose pateikiami 

duomenys privalo būti statistiškai įvertinti. Rodiklių žymėjimo santrumpos turi 

būti paaiškintos, jeigu jos neatitinka tarptautinių ISO standartų. 

Paveikslai 

Visa iliustracinė medžiaga – brėžiniai, grafikai, diagramos, fotografijos, piešiniai 

ir kt. – vadinami bendru paveikslų vardu. Tekstas juose rašomas lietuvių ir 

anglų kalbomis. 

Paveikslai turi būti nespalvoti, padaryti Microsoft Office 2000, 2003, XP, 

VISTA paketų elektroninėje lentelėje EXCEL su suderintomis programomis ir 

pradine informacija. Redakcija pasilieka teisę keisti jų formatą pagal straipsnio 

ar viso leidinio dizainą. 

Įrašai ir simboliai paveiksluose turi būti parašyti ne mažesniu kaip 10 dydžio 

TIMES NEW ROMAN šriftu. Paveikslų blokų dalys turi būti sužymėtos 

raidėmis a, b, c ir t. t. 

Literatūra 

1. 

2. 

Į literatūros sąrašą turi būti įtraukiamos tik mokslinės publikacijos. 

Citavimo pavyzdžiai: 

Vieno autoriaus knyga: 

Brazauskienė M. D. 2004. Agroekologija ir chemija. Naujasis lankas, 

Kaunas. 

Blažek J. 2001. Pìstujeme jablonì. Nakladatelství Brázda, s. r. o., Praha. 

Kelių autorių knyga: 

1. 

2. 

Kawecki Z., Łojko R., Pilarek B. 2007. Mało znane rośliny sadownicze. 

Wydawnictwo UWM, Olsztyn. 

Žemės ir miškų ūkio augalų pesticidų katalogas. 2005. G. Rimavičienė (sudaryt.). 

Lietuvos žemės ūkio konsultavimo tarnyba, Akademija, Kėdainių r. 

291

Straipsnis knygoje: 

1. Streif J. 1996. Optimum harvest date for different apple cultivar in the 

‘Bodensee’ area. In: A. de Jager, D. Johnson, E. Hohn (eds.), Determination and 

Prediction of Optimum Harvest Date of Apples and Pears. COST 94. European 

Commission. Luxembourg, 15–20. 

2. Byme D. H., Sherman W. B., Bacon T. A. 2000. Stone fruit genetic pool and 

its exploitation for growing under warm winter conditions. In: A. Erez (ed.), 

Temperature Fruit Crops in Warm Climates. Kluwer Academic Publishers, 

Netherlands, 157–230. 

3. Keller R. R. J. 2002. Cryopreservation of Allium sativum L. (Garlic). In: 

L. E. Towill, Y. P. S. Bajas (eds.), Biotechnology in Agriculture and Forestry. 

Springer-Verlag, Berlin, Heidelberg, 50: 38–47. 

1. 

1. 

1. 

2. 

Straipsnis konferencijos medžiagoje: 

Kampuss K., Strautina S. 2004. Evaluation of blackcurrant genetic resources 

for sustainable production. Proceedings of the International Workshop 

on Protection of Genetic Resources of Pomological Plants and Selection of 

Genitors with Traits Valuable for Sustainable Fruit Production. 22–25 August, 

Skierniewice, Poland, 147–158. 

Vieno autoriaus žurnalo straipsnis: 

Korban S. S. 1986. Interspecific hybridization in Malus. Hort. Science, 21(1): 

41–48. 

Žurnalo straipsnis (du ir daugiau autorių): 

Sasnauskas A., Gelvonauskienė D., Gelvonauskis B. 2002. Evaluation of 

new scab resistance apple in the first-fifth years in orchard. Sodininkystė ir 


Balan V., Oprea M., Drosu S., Chireceanu C., Tudor V., Petrisor C. 2006. 

Maintenance of biodiversity of apricot tree phenotypes in Romania. Acta 


Straipsnis e. žurnale: 

1. Stanys V., Mažeikienė I., Stanienė G., Šikšnianas T. 2007. Effect of phytohormones 

and stratification on morphogenesis of Paeonia lactiflora Pall. 

isolated embryos. Biologija, 18(1). http://images.katalogas.lt/maleidykla/ 

Bio71/Bio_027_030.pdf 

Duomenų bazė: 

FAO Stat Database. 2005. http://faostat.fao.org/faostat/ 

Disertacijos santrauka: 

1. Čižauskas A. 2003. Valgomųjų svogūnų ( Allium cepa L.) auginimo iš sėklų 

technologijos elementų tyrimai: daktaro disert. santr. Babtai. 

292

GUIDELINES FOR THE PREPARATION AND SUBMISSION 

OF ARTICLES TO THE VOLUMES OF SCIENTIFIC WORKS 

“SODININKYSTĖ IR DARŽININKYSTĖ“ 

Rules for Submission – Acceptance of Papers 

Papers can be contributed by any Lithuanian and foreign scientific workers 

and persons carrying out scientific research. The latter’s paper will be accepted 

only when the co-author is scientific worker. 

Manuscripts should be sent by mail typed on computer printed in two copies 

on paper taking in account following instructions. The manuscript will be registered 

and submitted to the member of the Editorial Board in charge. He (she) will 

evaluate if the contents and the form corresponds to the main requirements for 

periodical articles. Manuscripts rejected during the first evaluation will be returned 

to the author with explanatory remarks. If the article is approved the member of 

the Editorial Board appoints two reviewers. 

The author must return the corrected manuscript to the Editorial Board in ten 

days by e-mail or by mail in a diskette or CD. 

Requirements to the manuscript 

Structure and length 

The form of a manuscript has to correspond to the requirements for periodical 

scientific articles. The paper should be organized in the following order: 

– Title (should not exceed 10 words) 

Title should be written in small letters in bold (Aspects of plant color acclimation 

molecular mechanism). 

– Author(s)’ name, surname 

The name and surname should be written in small letters in bold. If there is 

more than one author they are listed according to their input to the paper. 

– Institution(s), address, email address 

Should be written in small letters in Italic (Lithuanian Institute of 

Horticulture). 

The main text 

– Abstract (should not exceed 1400 characters or 250 words) 

Should contain the statement of the aims, methods and main results in 

short. 

– Key words (should not exceed 10 words) 

– Introduction 

Should present the investigated subject, results of earlier related research, 

reasons of the study, innovation. Should indicate the aim of the investigation. 

293

– Object, methods and conditions 

– Results 

Should present concisely the collected data during investigation, documentation 

(tables, figures). 

– Discussion 

Should not repeat results presented in “Results” but should interpret them 

with reference to the results obtained by other authors, explain the reasons of the 

investigated phenomena and raise new ideas, hypotheses. 

– Conclusions 

– Acknowledgements (optional) 

– References 

Should be kept to a minimum of 10 latest references on this theme. 

– Summary in Lithuanian and English (600–1 400 characters) 

Articles written based on non-traditional trial data and the obtained results 

may have other than traditional structural parts of a paper. 

The article should not exceed 10 pages of computer text, tables and figures 

included (longer articles are agreed with the chairman of the Editorial Board). 

Text preparation 

The manuscripts should be submitted in Lithuanian and English, typed on a 

PC, used Microsoft WORD for Windows 2000 XP or VISTA word-processor 

format. The font to be typed – TIMES NEW ROMAN size 12, on A4 paper 

(210 × 297 mm), single spaced, justified. Margins: top – 2 cm, bottom – 2 cm, 

right – 1.5 cm, left – 3 cm. Article should be submitted in electronic carrier. 

In bold there are written the title of the paper in all languages, headings and all 

main structural elements (introduction, object, methods and conditions, results, 

discussion, conclusions, acknowledgements, references, abstract). In Italic there 

are written Latin names of species, genera, diseases, pests micro-organisms and 

other biological objects. Names of plant cultivars should be placed within single 

quotation marks (for example, ‘Auksis’). 

The quoted reference in the text is indicated in round brackets (author’s surname, 

year). 

Tables 

Information given in figures or other illustrations, should not be repeated in 

tables. 

Text in tables is written in Lithuanian and English languages. If Lithuanian 

and English texts are in one line they are separated by /. Do not use vertical and 

horizontal lines to separate parts of the text. A horizontal line separates only headings 

of columns and the end of the table. Orientation in a page only vertical 

(Portrait). 

Trial variants in tables should not be numbered or submitted in complicated 

abbreviations. Tables should be self explanatory, and if there are abbreviations, 

294

they should be understandable. The used abbreviation should be explained in the 

description of first table (figure). 

Statistical data, figures, numerals 

It is desirable to describe in detail the applied research methods and indicate 

their original references. The information on the scheme (design) of field, 

vegetative and other trials and motivation of their choice is very important. Data 

presented in tables and figures must be statistically processed. Abbreviations 

of parameters should be explained if they do not correspond to the international 

standard abbreviations (ISO). 

Figures 

All illustrations – drawings, graphs, diagrams, photographs, pictures, etc. are 

considered as figures. The text in them is written in Lithuanian and English. 

Figures must be drafted in black colour in Microsoft Office 2000, 2003, XP, 

VISTA package, EXCEL electronic table with conformed programs and initial 

information. Editorial Board has the right to change their format according to the 

design of the article or the whole publication. 

Letters and symbols in figures should be not smaller than size 10 TIMES 

NEW ROMAN. Block parts of figures should be numbered consecutively by 

letters a, b, c, etc. 

References 

Into references it may be included scientific publications. 

Titles of foreign journals, volumes of conference articles, etc., are not abbreviated. 

The reference list should be arranged in alphabetical order. 

Examples of quotation: 

Book of one author: 

1. 

2. 

1. 

2. 

Brazauskienė M. D. 2004. Agroekologija ir chemija. Naujasis lankas, 

Kaunas. 

Blažek J. 2001. Pìstujeme jablonì. Nakladatelství Brázda, s. r. o., Praha. 

Books of several authors: 

Kawecki Z., Łojko R., Pilarek B. 2007. Mało znane rośliny sadownicze. 

Wydawnictwo UWM, Olsztyn. 

Žemės ir miškų ūkio augalų pesticidų katalogas. 2005. G. Rimavičienė (sudaryt.). 

Lietuvos žemės ūkio konsultavimo tarnyba, Akademija, Kėdainių r. 

295

Article in book: 

1. Streif J. 1996. Optimum harvest date for different apple cultivar in the 

‘Bodensee’ area. In: A.de Jager, D. Johnson, E. Hohn (eds.), Determination 

and Prediction of Optimum Harvest Date of Apples and Pears. COST 94. 

European Commission. Luxembourg, 15–20. 

2. Byme D. H., Sherman W. B., Bacon T. A. 2000. Stone fruit genetic pool and 

its exploitation for growing under warm winter conditions. In: A. Erez (ed.), 

Temperature Fruit Crops in Warm Climates. Kluwer Academic Publishers, 

Netherlands, 157–230. 

3. Keller R. R. J. 2002. Cryopreservation of Allium sativum L. (Garlic). In: 

L. E. Towill, Y. P. S. Bajas (eds.), Biotechnology in Agriculture and Forestry. 

Springer-Verlag, Berlin, Heidelberg, 50: 38–47. 

Article in conference material: 

1. 

1. 

2. 

3. 

1. 

Kampuss K., Strautina S. 2004. Evaluation of blackcurrant genetic resources 

for sustainable production. Proceedings of the International Workshop 

on Protection of Genetic Resources of Pomological Plants and Selection of 

Genitors with Traits Valuable for Sustainable Fruit Production. 22–25 August, 

Skierniewice, Poland, 147–158. 

Article in scientific journal: 

Korban S. S. 1986. Interspecific hybridization in Malus. HortScience, 21(1): 

41–48. 

Sasnauskas A., Gelvonauskienė D., Gelvonauskis B. 2002. Evaluation of 

new scab resistance apple in the first-fifth years in orchard. Sodininkystė ir 


Balan V., Oprea M., Drosu S., Chireceanu C., Tudor V., Petrisor C. 2006. 

Maintenance of biodiversity of apricot tree phenotypes in Romania. Acta 


Article in e. journal: 

Stanys V., Mažeikienė I., Stanienė G., Šikšnianas T. 2007. Effect of phytohormones 

and stratification on morphogenesis of Paeonia lactiflora Pall. 

isolated embryos. Biologija, 18(1). http://images.katalogas.lt/maleidykla/ 

Bio71/Bio_027_030.pdf 

Database: 

FAO Stat Database. 2005. http://faostat.fao.org/faostat/ 

Thesis abstract: 

Čižauskas A. 2003. Valgomųjų svogūnų ( 

1. Allium cepa L.) auginimo iš sėklų 

technologijos elementų tyrimai: daktaro disert. santr. Babtai. 

296

Contents – Turinys 

V. Arnaudov, H. Kutinkova 

Controling pear psylla with Abamectin in Bulgaria....................................................3 

Kriaušių blakučių naikinimas Abamektinu Bulgarijoje..............................................9 

G. Bimsteine, L. Lepse, B. Bankina 

Possibilities of integrated management of onion downy mildew..............................11 

Svogūnų netikrosios miltligės integruotos kontrolės galimybės...............................17 

D. Bridžiuvienė, J. Repečkienė 

Interspecific relation peculiarities between soil and phytophatogenic fungi ........... 19 

Dirvožemio ir fitopatogeninių mikromicetų tarprūšinės sąveikos ypatumai............ 28 

O. Bundinienė 

Influence of boron fertilizer and meteorological conditions on red beet infection 

with scab and productivity ........................................................................................29 

Boro trąšų ir meteorologinių sąlygų įtaka burokėlių užsikrėtimui rauplėmis ir 

derlingumui................................................................................................................40 

L. Duchovskienė, L. Raudonis, R. Karklelienė, R. Starkutė 

Toxicity of insecticides to predatory mite Phytoseuilus persimilis in cucumber...... 41 

Insekticidų toksiškumas grobuoniškoms erkėms Phytoseuilus persimilis agurkuose... 

...................................................................................................................................46 

L. Duchovskienė, E. Survilienė 

Effect of Abamectin on two-spotted spider mite and leaf miner flies in greenhouse 

cucumbers......................................................................................................47 

Abamektino poveikis paprastosioms voratinklinėms erkėms ir minamusėms 

šiltnamio agurkuose...................................................................................................56 

M. Eihe, R. Rancane, L. Vilka 

Different fungicide combinations against apple scab helping to avoid fungus 

resistance .................................................................................................................57 

Įvairūs fungicidų deriniai nuo obelų rauplių, padedantys išvengti rauplėgrybio 

atsparumo..................................................................................................................67

J. Jankauskienė, E. Survilienė 

Influence of growth regulators on seed germination energy and biometrical 

parameters of vegetables...........................................................................................69 

Augimo reguliatorių įtaka daržovių sėklų dygimo energijai ir daigų vystymuisi......... 

...................................................................................................................................77 

S. Jarmoliča, B. Bankina 

Powdery mildew of strawberries in Latvia under field conditions............................79 

Braškių netikroji miltligė Latvijoje lauko sąlygomis................................................83 

K. Jõgar, L. Metspalu, K. Hiiesaar, L. Loorits, 

A. Ploomi, A. Kuusik, A. Luik 

Influence of neemazal-T/S on Mamestra brassicae L.............................................85 

Nimazalio T/S poveikis Mamestra brassicae L........................................................92 

V. Kamardzina 

Sensitivity of Venturia inaequalis populations to krezoxym methyl.........................93 

Venturia inaequalis populiacijų jautrumas krezoksym metilui ..............................100 

D. Kavaliauskaitė 

Efficacy of herbicide Lentagran WP for control of annual dicotyledonous weeds in 

cabbage crop ...........................................................................................................101 

Herbicido Lentagran WP veiksmingumas kopūstų pasėlyje naikinant vienametes 

dviskiltes piktžoles .................................................................................................108 

D. Kviklys 

Tolerance of apple propagation material to herbicides ...........................................109 

Obelų dauginamosios medžiagos tolerantiškumas herbicidams ............................115 

N. Kviklienė, A. Valiuškaitė 

Influence of maturity stage on fruit quality during storage of ‘Shampion’ apples........ 

.................................................................................................................................117 

Skynimo laiko įtaka ‘Šampion’ obuolių kokybei vaisiams nokstant ir juos laikant...... 

.................................................................................................................................123 

V. Laugale, L. Lepse, L. Vilka, R. Rancāne 

Incidence of fruit rot on strawberries in Latvia, resistance of cultivars and impact 

of cultural systems .................................................................................................125 

Vaisių puvinio paplitimas ant braškių Latvijoje, veislių atsparumas ir kultūrinių 

sistemų įtaka............................................................................................................134

R. Mikaliūnaitė, M. Kazlauskas, L. Veriankaitė 

Prevalence peculiarities of airborne Alternaria genus spores in different areas 

of Lithuania.............................................................................................................135 

Ore plintančių Alternaria genties grybų sporų sklaidos ypatumai skirtingose 

Lietuvos vietovėse ..................................................................................................142 

L. B. Orlikowski, M. Ptaszek, A. Trzewik, T. Orlikowska 

Water as the source of Phytophthora spp. pathogens for horticultural plants............... 

.................................................................................................................................145 

Vanduo kaip sodo augalų ligų sukėlėjo Phytophthora spp. šaltinis........................151 

A. Ploomi, K. Jõgar, L. Metspalu, K. Hiiesaar, 

L. Loorits, I. Sibul, I. Kivimägi, A. Luik 

The toxicity of Neem to the snail Arianta arbustorum...........................................153 

Neem toksiškumas medsraigei Arianta arbustorum ..............................................158 

M. Ptaszek, L. B. Orlikowski, C. Skrzypczak 

New host plants for development of Phytophthora cryptogea in Poland...............159 

Nauji augalai Phytophthora cryptogea vystymuisi Lenkijoje.................................164 

N. Pūpola, L. Lepse, A. Kāle, I. Moročko-Bičevska 

Occurrence of RBDV in Latvia and virus elimination in vitro by chemotherapy......... 

.................................................................................................................................165 

Aviečių žemaūgiškumo viruso (RBDV) paplitimas Latvijoje ir viruso naikinimas 

in vitro chemoterapija..............................................................................................172 

L. Raudonis, A. Valiuškaitė, L. Duchovskienė, E. Survilienė 

Toxicity of biopesticides to green apple aphid in apple-tree...................................173 

Biopesticidų toksiškumas žaliesiems obeliniams amarams....................................179 

L. Raudonis, A. Valiuškaitė 

Integrated aproach of apple scab management using iMETOS warning system ...181 

Integruotas obelų rauplių valdymas naudojant iMETOS prognozavimo sistemą......... 

.................................................................................................................................191 

R. Rodeva, J. Gabler, Z. Stoyanova 

First evidence of Itersonilia perplexans on dill (Anethum graveolens) in Bulgaria..... 

.................................................................................................................................193 

Pirmieji Itersonilia perplexans požymiai ant krapų (Anethum Graveolens) 

Bulgarijoje...............................................................................................................198

M. Samuitienė, M. Navalinskienė, S. Dapkūnienė 

Investigation of Tobacco rattle virus infection in peonies (Paeonia L.).................199 

Tabako garbanotosios dryžligės (Tobacco rattle virus, TRV) viruso infekcijos 

bijūnuose (Paeonia L.) tyrimas...............................................................................208 

A. Sasnauskas, D. Gelvonauskienė 

Evaluation of agronomical characters and resistance to fungal diseases of apple 

cultivars...................................................................................................................209 

Obelų veislių agronominių požymių ir atsparumo grybinėms ligoms tyrimas ........216 

R. Starkutė, L. Duchovskienė, V. Zalatorius 

Influence of preplant and vegetable crop rotation links on carrot yield and damage 

of pests ....................................................................................................................217 

Priešsėlių ir daržovių sėjomainos grandžių įtaka morkų derliui ir kenkėjų daromiems 

pažeidimams ...........................................................................................................224 

E. Survilienė, A. Valiuškaitė, V. Snieškienė, A. Stankevičienė 

Effect of essential oils on fungi isolated from apples and vegetables.....................227 

Eterinių aliejų poveikis grybams, išskirtiems iš obuolių ir daržovių......................234 

E. Survilienė, L. Raudonis, J. Jankauskienė 

Investigation of pesticides effect on pollination of bumblebees in greenhouse 

tomatoes...................................................................................................................235 

Pesticidų poveikio kamanių entomofilijai šiltnamio pomidoruose tyrimas............241 

A. T. Wojdyła 

Effictiveness of Olejan 85 EC against chrysanthemum and willow rust.................243 

Olejan 85 EC veiksmingumas kontroliuojant chrizantemų ir gluosnių rūdis ........248 

A. Yankouskaya 

Application of biological insecticide Pecilomicine-B for greenhouse pest control .....249 

Biologinio insekticido Pecilomicine-B poveikis šiltnamio kenkėjams .................258 

S. Yarchakovskaya, N. Kaltun 

Optimization of time and expediency of Incurvaria capitella Cl. number 

regulation.................................................................................................................259 

Incurvaria capitella Cl. kontroliavimo laiko ir tikslingumo optimizavimas .........268

I. Zarins, M. Daugavietis, J. Halimona 

Biological activity of plant extracts and their application as ecologically harmless 

biopesticide..............................................................................................................269 

Augalų ekstraktų biologinis efektyvumas ir jų naudojimas kaip ekologiškai nekenksmingų 

biopesticidų..................................................................................................280 

I. Zitikaitė, M. Samuitienė 

Detection and characterization of Cucumber mosaic virus isolated from sweet 

peppers.....................................................................................................................281 

Iš saldžiųjų paprikų izoliuoto agurkų mozaikos viruso nustatymas ir charakterizavimas....................................................................................................................288 

Atmintinė autoriams rašantiems į Mokslo darbus „Sodininkystė ir daržininkystė“ ..... 

.................................................................................................................................289 

Guidelines for the preparation and submission of articles to the volumes of scentific 

works “Sodininkystė ir daržininkystė”....................................................................293

ISSN 0236-4212 

Mokslinis leidinys 

Lietuvos sodininkystės ir daržininkystės instituto ir 

Lietuvos žemės ūkio universiteto mokslo darbai 

Sodininkystė ir daržininkystė. T. 28(3). 1–304. 

Redagavo ir skaitė korektūrą Jolanta Kriūnienė 

Kompiuteriu maketavo Larisa Kazlauskienė 

SL 1070. 2009 08 27 19 sp. l. Tiražas 200 egz. 

Išleido Lietuvos sodininkystės ir daržininkystės institutas, 

LT-54333 Babtai, Kauno r. 

Spausdino UAB „Spaudos brokeris“ Vytauto pr. 27, 

LT-44352 Kaunas 

Užsakymo Nr. 9184

Volume / tomas 28(3) - SodininkystÄ ir darÅ¾ininkystÄ - SodininkystÄs ...

Create successful ePaper yourself

Delete template?

Save as template?

Volume / tomas 28(3) - SodininkystÄ ir darÅ¾ininkystÄ - SodininkystÄs ...