Vol.12_No.2 - Pesticide Alternatives Lab - Michigan State University

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Spring 2003 Resistant Pest Management Newsletter Vol. 12, No.2followed by population from Nalgonda (9.13 µg/µL),Guntur (8.73), Mysore (5.15), and Dharwad (4.84). Thelowest LD50 value was observed in the populationfrom Madurai (0.78µg/µL) followed by Kovilpatti(0.87), Coimbatore (1.35), and Madhira (1.12). Theresistance ratio (RR) against the ICRISAT susceptiblestrain was found to be highest for the population ofRaichur (66.8 fold) followed by Nalgonda (45.7),Guntur (43.6), Mysore (25.8), Dharwad (24.2), Nagpur(14.6), and Nanded (10.7). The least resistance ratiowas observed in the population of Madurai (3.9)followed by Kovilpatti and Madhira (Table 6).The general LD50 values recorded were far highercompared to the recommended dosages indicating theexistence of resistance as was reported earlier (Armeset al., 1992). However, the resistance drastically differsfrom location to location within the South Indiancotton ecosystems. The resistance levels in the Guntur,Nalgonda, and Raichur regions (heavy insecticideusage areas) are due to heavy dependence oninsecticides. This clearly explained that resistancelevels were proportionate with the usage of pesticides.The study conducted by Forrester (1990) also clearlyrevealed that resistance levels rose when pyrethroidswere used but fell significantly when they werewithheld. Thus, the pesticides were creating very highselection pressure for resistant genotypes. Thissuggests that indiscriminate use and heavy dependenceon pesticide will further complicate the alreadyworsened situation and hints at aiming for insecticideresistance management strategies.ACKNOWLEDGEMENT: We thank Dr. Daniel R. Zeigler,Ohio State University, US for providing the Cry1Acover expressing clone. This research work wassupported by the DBT, GOI project grants to BF.Thanks are also due to Dept. of Agril. Entomology,UAS Dharwad for providing infrastructure facilitiesand all those scientists who helped us during the visitto different locations across south India for H.armigera collection. We thank Dr. G.T.Gujar, IARI,New Delhi, for his useful suggestions during the studyand encouragement.REFERENCES:KRANTHI.K.R., JADHAV,D.R., WANJARI,R.R., SHAKHIR ALI,S.AND RUSSEL,D., 2001, Carbamate and organophosphateresistance in cotton pests in India. Bull.Ent. Res., 91:37-46.Mc CAFFEREY, A.R., KING, A.B.S., WALKER, A.J. AND EL-NAYIR, H., 1989, Pestic.Sci., 27:65-76.RAKILA, A. AND PADMANABHAN, N.R., 1995,Knowledge andfactor influencing pesticide use and frequency of plant protectionmeasures. Pestology, 19:9-12. ROSS, G.J.S., 1987,Maximumlikelihood programme. The Numerical Algorithm groupRothemsted Experimental Station, Harpendon,U.K.ARMES, N.J., JADHAV, D.R., BOND, G.R. AND KING, A.B.S., 1992,Insecticides resistance in Helicoverpa armigera in South India.Pestic Sci., 34:335-364.FORRESTER, N.W., 1990, Designing, Implementing and Servicing anInsecticide resistance Management Strategy. Pestic sci., 28:167-179.B. Fakrudin*, Badariprasad, K. B. Krishnareddy, S. H.Prakash, Vijaykumar, B.V. Patil#, & M. S.KuruvinashettiDepartment of BiotechnologyUniversity of Agricultural Sciences DharwadKrishinagar, Dharwad - 580005, KarnatakaIndia#Professor and HeadDept. of Agricultural EntomologyCollege of AgricultureRaichur - 584101, KarnatakaIndia*CorrespondenceDevelopment of Resistance in Insects to Transgenic Plants with Bacillus thuringiensis Genes: Current Status andManagement StrategiesINTRODUCTION There is a continuing need to increasefood production as the world population is expected toexceed 6 billion by 2050. In both the developed andundeveloped countries, the cost for achievingproduction has become too high because of the need toincur costs for controlling insect pests that cause anestimated loss of $10 billion annually. The difficultiesexperienced in controlling insect pests over the past 30years have been largely due to the over-use ofpesticides. Indiscriminate use of insecticides has ledboth to the development of resistance in insects and thedestruction of natural enemies. Bio-pesticides such asBt (Bacillus thuringiensis) products are widelyregarded as being the least harmful to natural enemies.Because of its selectivity and environmental safety,usage of Bt is increasing, particularly in IPM programs.Foliar application of Bt breaks down quickly underfield conditions due to UV sensitivity and rainfall.With the advent of recombinant DNA technologies,insecticidal proteins present in Bt have been expressedin crop plants to ensure durable insect resistance. Thereis a considerable increase in global area undertransgenic crops from 1.7 million hectares in 1996 to52.6 million hectares in 2001, in which the share of Btcrops was 15% of the total area (James 2001).Although much progress has been made in thediscovery of new genes for introduction into plants,only Bt genes have been exploited so far.Considerable efforts have been made toincorporate delta-endotoxins from Bt into cereals, rootcrops, leafy vegetables, forage crops, and horticulturalcrops (Schuler et al. 1998). Of the $8.1 billion (US16
Spring 2003 Resistant Pest Management Newsletter Vol. 12, No.2dollars) spent annually on insecticides worldwide, itwas estimated that nearly $2.7 billion could besubstituted with Bt biotechnology applications(Krattiger 1997). Economic advantage gained during1999 by Bt cotton alone has been estimated to be $213million in the USA. Cultivation of transgenic crops hasled to a reduction in pesticide use and significantincrease in yield (Cannon 2000). Unfortunately, thereare also concerns that the benefits of geneticallytransformed plants will be short-lived (McGaughey &Whalon 1992). Despite the potential advantages ofusing Bt crops, the possibility of their widespread usehas raised some potential problems. Decades ofindiscriminate insecticide use have demonstrated thatexposing insect populations to high levels of toxinsresults in evolution of resistance to insecticides (Roush& McKenzie 1987). Recently, several species of insectpests have been selected for resistance to Bt in thelaboratory, indicating that biological pesticides cansuffer the same fate as the chemical pesticides (Lianget al. 2000, McGaughey et al. 1998a).DEVELOPMENT of RESISTANCE in INSECTS to Bt GENESSeveral studies have shown that insect pests can adaptto Bt toxins under laboratory conditions (Shelton et al.2002). Certain pests such as Plodia interpunctella(McGaughey 1985), Heliothis virescens (Stone et al.1989), Plutella xylostella (Tabashnik et al. 1990),Spodoptera exigua (Moar et al. 1995), and Ostrinianubilalis (Huang et al. 1997) have been shown todevelop some degree of resistance to B. thuringiensisunder laboratory conditions. Evolution of insectresistance to insecticidal proteins produced by Btwould decrease our ability to control agricultural pestswith genetically engineered crops designed to expressgenes coding for these proteins (Gould et al. 1992).Information on development of resistance in insects toBt toxins has been summarized below.Indian meal moth, Plodia interpunctella:The first studied case of resistance to Bt-strainswas P. interpunctella, which had developed 100-foldresistance following 15 generations of laboratoryselection with Dipel (McGaughey 1985). On furtherselection, after 36 generations, the resistance levelsreached 250-fold (McGaughey & Beeman 1988).Bacillus thuringiensis sub sp. kurstaki caused a narrowspectrum resistance to Cry1Ab and Cry1Ac toxins,while sub sp. aizawai and entomocidus strains causedbroad-spectrum resistance to Cry1Aa, Cry1Ab,Cry1Ac, Cry1B, Cry1C, and Cry2A (McGaughey &Johnson 1994).Diamondback moth, Plutella xylostellaAlthough there are no instances of insectsdeveloping resistance to Bt transgenic plants in thefield, diamondback moth, P. xylostella, is the firstinsect known to have evolved high levels of resistanceto Bt as a result of repeated use of formulated Btinsecticide (Tabashnik et al. 1990). A diamondbackmoth colony derived from field population in thePhilippines that was regularly exposed to Dipel showedmore than 200-fold resistance to Cry1Ab (Ferre et al.1991). As much as 1640-fold resistance to Bt has beenrecorded in localized populations of diamondbackmoth from Hawaii, Florida, and Asia (Tabashnik et al.1992). In field populations of P. xylostella, resistanceto Bt sub sp. kurstaki, containing Cry1A(a,b,c), Cry2A,and Cry2B toxins and to a lower extent Bt sub sp.aizawai, containing Cry1A (a,b), Cry1C, and Cry1Dtoxins has been observed in various countries(Tabashnik 1994). Laboratory selection of P. xylostellausing Cry1Ca protein and in later generation transgenicbroccoli expressing Cry1Ca, increased Cry1Caresistance to 12400-fold (Zhao et al. 2000b).Resistance to Cry1A toxins from Bt sub sp. kurstakicaused cross-resistance to Cry1F, but not to Cry1B orCry1C (Tabashnik et al. 1996). Contrary to theassumption that independent mutations are required tocounter each toxin in P. xylostella, an autosomalrecessive gene conferred extremely high resistance toCry1Aa, Cry1Ab, Cry1Ac, and Cry1F (Tabashnik et al.1997). In a P. xylostella colony possessing 1500-foldresistance to a commercial formulation, the resistancerapidly fell to 300-fold in the absence of selection, butremained stable at this level in subsequent generations(Tang et al. 1996).Cotton bollworm/ legume podborer, Heliothis/HelicoverpaHelicoverpa armigera is capable of developingresistance to Cry1Ac in 7 to 8 generations (Kranthi etal. 2000). Highly mobile polyphagous pests such asHelicoverpa may develop resistance to Bt on onetransgenic crop and then disperse, nullifying theeffectiveness of a wide range of Bt transgenic cropsexpressing the same or similar Cry proteins. Pests withresistance to CryIA proteins in transgenic plants mayalso display significant resistance to Bt biopesticides.A laboratory strain of H. virescens developedresistance in response to selection with the Bt toxinCryIAc. In contrast to other cases of Bt-toxinresistance, this strain exhibited cross-resistance to Bttoxins that differ significantly in structure and activity(Gould et al. 1992). Over 10000-fold resistance toCry1Ac was obtained in H. virescens colony onselection with Cry1Ac protoxin (Gould et al. 1995).The insecticidal activity of Bt in leaves and squares oftransgenic cotton plant was high during the secondgeneration of the insect, but declined in the third andfourth generations of H. armigera in North China. Thesurviving third and fourth generation larvae, afterfeeding on flowers of Bt cotton, fed on the bolls untilpupation, which caused selection in field populationsof H. armigera. The increase in resistance was 7.1-fold17
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