RAPD-based assessment of genetic diversity among annual ...

EurAsian Journal of BioSciences 

Eurasia J Biosci 5, 37-47 (2011) 

DOI:10.5053/ejobios.2011.5.0.4 

RAPD-based assessment of genetic diversity among 

annual caraway (Carum carvi) populations 

Bochra Laribi1*, Nejia Zoghlami2, Myriam Lamine2, Karima Kouki1, Abdelwahed Ghorbel2, Abdelaziz Mougou1 1Department of Agronomy and Vegetal Biotechnology, National Agronomic Institute 

of Tunisia, 1082 Tunis, Tunisia 

2Laboratory of Plant Molecular Biology, Center of Biotechnology of Borj Cedria, 

BP 901, 2050 Hammam-Lif, Tunisia 

*Corresponding Author: bochra_laribi@yahoo.fr 

Abstract 

For the first time, genetic variability and differentiation among five annual caraway (Carum 

carvi) populations originating from Tunisia, Germany and Egypt were examined. Random 

Amplified Polymorphic DNA (RAPD) marker data were obtained and analysed with respect to 

genetic diversity, population structure and gene flow. Fourteen primers generated a total of 

136 discernible and reproducible bands across the analyzed populations, out of which 56 were 

polymorphic. The UPGMA cluster analysis permitted the discrimination of all the genotypes and 

their sorting into 3 main groups. Tunisian caraway populations diverged significantly from 

German and Egyptian ones. Population clustering was made dependently from geographic 

origin. This has been further explained at the DNA level as we were able to select a set of 

RAPD fingerprints unique to each of the studied populations. Furthermore, dimensional graph 

derived from factorial analysis of RAPD frequency data, allowed significant grouping of the 

genotypes into five sub-plots, representing each one population. Shannon's index values 

showed that variation ranks between rather than within populations. These results indicated 

that considerable genetic differences among C. carvi populations were registered. 

Keywords: Carum carvi, differentiation, diversity, population. 

Laribi B, Zoghlami N, Lamine M, Kouki K, Ghorbel A, Mougou A (2011) RAPD-based 

assessment of genetic diversity among annual caraway (Carum carvi) populations. Eurasia J 

Biosci 5: 37-47. 

DOI:10.5053/ejobios.2011.5.0.5 

INTRODUCTION 

Caraway (Carum carvi L.) is one of the 

oldest aromatic and medicinal plants of the 

Apiaceae family which is native to Asia, 

Europe and North Africa (Nemeth 1998). It is 

mainly cultivated in the Netherlands, Finland, 

Hungary, Morocco, Iran, India and Russia 

(Hornok 1986, Toxopeus and Bowmeester 

1993). Besides it is being used as a 

condiment, caraway has been utilized for a 

long time in traditional medicine especially for 

the treatment of digestive disorders and was 

commonly used in phytomedicine as diuretic 

(Lahlou et al. 2007), anti-hyperglycaemic 

(Eddouks et al. 2004, Ene et al. 2007, 

Tahraoui et al. 2007), anti-hyperchlolesterol 

(Lemhadri et al. 2006) and anti-cancerous 

(Naderi-Kalali et al. 2005, Kamaleeswari et al. 

2006). 

Little is known about caraway genetic 

diversity and population structure despite its 

importance as an aromatic and medicinal 

plant. Only a few reports based on the species 

of Lithuania are available and relevant to the 

diversity and ex situ stability of 

morphological, biochemical and productivity 

traits of C. carvi cenopopulations growing 

wild in natural habitats such as grasslands 

(Petraityte 2003, Petraityte and Dastikaite 

2007, Dabkevicius et al. 2008). Thus, with 

the decline of these lasts, mainly by 

anthropogenic activities, the C. carvi 

biodiversity and genetic resources become 

sparse (Petraityte 2003, Petraityte and 

Dastikaite 2007). 

Received: February 2011 

Received in revised form: May 2011 

Accepted: June 2011 

Printed: June 2011 

©EurAsian Journal of BioSciences, 2011 37


During the last few years, the 

characterization and evaluation of genetic 

diversity and relationships within and between 

species and populations were performed 

generally by molecular techniques that 

substituted the classic ones such as 

morphological and physiological characters 

(Dolezalova et al. 2003). 

Indeed, the nucleotide sequences of the 

internal transcribed spacer region were used 

as molecular markers in order to establish 

phylogenetic relationships of some Apiaceae 

genera like Chamaesciadium C. A. Meyer 

(Papini 2006) and Carum L. (Papini et al. 

2007). It have been demonstrated that this 

last genus is polyphyletic. Indeed, maximum 

parsimony with bootstrap resampling, 

maximum likelihood and Bayesian inference 

analyses resulted in three distinct clades for 

this genus (Papini et al. 2007). 

In addition, the RAPD remains one of the 

most extensively used molecular techniques 

due to its simplicity, low cost and high speed. 

Thus, RAPD markers have been successfully 

used in many crops in providing a convenient 

and rapid assessment of genetic diversity 

among different genotypes (Williams et al. 

1990, Rafalski and Tingey 1993, Ragot and 

Hoisington 1993). In general, RAPD can 

provide valuable data in the analysis of 

population genetic structure including genetic 

diversity within and among populations, 

population subdivision, and degree of 

inbreeding and individual relatedness (Lunch 

and Milligan 1994). 

In case of aromatic and medicinal plants, 

RAPD markers have been scarcely used in 

genetic diversity studies, e.g. in Origanum 

majorana L. (Klocke et al. 2002), Cunila 

galioides Benth (Fracaro et al. 2005), 

Curcuma zedoaria (Christm.) Rosc. (Islam et 

al. 2007, Syankumar and Sasikuma 2007), 

Matricaria chamomilla L. (Solouki et al. 2008), 

Satureja hortensis L. (Hadian et al. 2008) and 

Foeniculum vulgare Mill. (Zahid et al. 2009). 

However, molecular data regarding Tunisian 

caraway genetic resources still have been 

missing. Thus, in this study we present the 

first survey of genetic diversity among five C. 

carvi populations; three from Tunisia, one 

38 

Laribi et al. 

from Germany and the other from Egypt, 

detected with molecular RAPD markers. This 

work aims at investigating the extent of gene 

flow within and between these populations of 

C. carvi as a first step to provide baseline 

information for the development of strategies 

for the conservation and breeding of this 

aromatic and medicinal plant. 

MATERIAL AND METHODS 

Plant material and growth conditions 

Five Carum carvi L. populations of 

cultivated origin were used in this study. 

Three populations were collected from the 

field in the principal production regions of 

caraway in Tunisia which are Haouaria, 

Menzel Temime and Souassi. The two others 

were imported, one from Germany and the 

other from Egypt (Table 1). 

Seeds were sown in pots of 1.5 L volume 

filled by agricultural soil containing 0.22, 

0.34, 0.05 and 0.08 mequiv. (100 g -1) of dry 

soil of Na +, K +, Ca 2+, Fe 2+, respectively. The 

pots were kept in a greenhouse at day light 

(photoperiod varying from 13 to 16 h) and at 

a temperature varying between 18 and 20°C 

during the day and between 10 and 12°C 

during the night at the Centre of 

Biotechnology of Borj Cedria (36°41'47 N, 

10°29'30 E, 26 m above sea level). Preirrigation 

was done immediately after sowing 

and the moisture content of the soil was 

controlled by regular irrigation intervals 

according to weather conditions for uniform 

emergence and seedlings establishment which 

took place approximately four weeks after 

sowing. After that, fresh leaves were sampled 

separately from 10 plants per population on 

the basis of one plant per pot, and were 

immediately ground to a powder in liquid 

nitrogen and then stored at -80°C. 

DNA extraction 

Total foliar DNA was extracted following 

the protocol of Bowers et al. (1993) later 

modified by This et al. (1997) and Zoghlami et 

al. (2001). Extracted DNA was quantified by 

visual comparison with a molecular size 

marker (1 kb ladder) on ethidim bromide 

stained agarose gels. 

©EurAsian Journal of BioSciences, 2011


DNA amplification 

Seventeen universal decamer oligonucleotides, 

purchased from the University of 

British Colombia were used for the 

polymerase chain reaction (PCR) amplification 

(Table 2). They were tested on three 

populations for their ability to produce 

polymorphic, unambiguous and stable RAPD 

markers (Table 2). 

DNA amplification reactions and electrophoresis 

were performed according to Zoghlami 

et al. (2007). Amplifications were repeated at 

least twice and only reproducible products 

were taken into account for further data 

analysis. 

Data analysis 

Polymorphic DNA bands were screened 

and photographed by a Biometra Bio-doc-IITM 

system (Biometra, Göttingen, Germany). 

Bands were carefully selected from replicated 

amplifications in order to ensure the absence 

of artefacts and were designated by their 

primer code and their size in base pairs. 

Polymorphic DNA bands were recorded as 

discrete variables for the presence (1) or for 

the absence (0) of a similar band. 

For each primer, the number of bands and 

the polymorphic ones were calculated as well 

as the percentage of polymorphic bands 

(PPB). The latter was determined as the 

percentage of polymorphic bands over the 

total number of the yielded bands. The 

number of RAPD banding profiles (profiles 

generated by all the populations per primer) 

has also been calculated. The ability of the 

most informative primers to differentiate 

between populations was assessed by the 

estimation of their resolving power (Rp) 

(Prevost and Wilkinson 1999). The Rp has 

been described to correlate strongly with the 

ability to distinguish between the populations 

according to the following Gilbert et al. 

(1999) formula: 

Rp= Ib 

Where Ib= 1- ( ) where p is the 

population proportion containing the I band. 

RAPD bands were transformed into a 

binary character matrix. Populations were 

then clustered using the Unweighed Pair 

Group Method with Arithmetic averaging 

Laribi et al. 

algorithm (UPGMA, Sneath and Sokal 1973) 

using Darwin software (Version 5.0.148, 

http://darwin.cirad.fr/darwin). The origin of 

populations under investigation was taken 

into account in order to examine their 

potential effect on the genetic clustering. We 

also investigated genetic variation among 

populations using Factorial Correspondence 

Analysis (FCA). 

Population differentiation analysis has been 

assessed among and within the studied 

populations according to Nei (1978) using the 

computer program POPGENE 1.32 (Yeh et al. 

1999). Shannon's index of genotypic diversity 

(H0) for RAPD was estimated to quantify the 

degree of within-population diversity following 

the calculation: 

H0=- piln pi ; where pi was the frequency 

of presence or absence of a RAPD band in a 

population, as described by Yeh et al. (1995). 

The average diversity over all populations 

(Hpop) was calculated as: 

1 

H H 

pop= o 

n ; where n was the number of 

populations Yeh et al. (1995). 

The mean diversity at species level (Hsp) was calculated as: 

Hsp=- psln ps; where ps was the frequency 

of presence or absence of a RAPD 

band in all the populations. 

The component of within-population 

diversity was estimated as Hpop/Hsp, and that 

of between-population diversity as 1-Hpop/Hsp. The gene flow (Nm, number of migrants per 

generation) (McDermott and McDonald 1993) 

was approximated as: 

Nm= 0.5*(1-GST)/GST; where GST (analogous 

to FST) (Wright 1951) was the diversity 

coefficient among populations. 

RESULTS 

Genetic polymorphism 

A total of 17 primers were screened for 

their ability to generate consistently amplified 

band patterns and to assess polymorphism in 

the tested populations (Table 2). One hundred 

and thirty six (136) bands were obtained, 

which is an average of 9.7 bands per primer: 



Table 1. Origin of C. carvi populations included in 

this study with their respective geographic 

coordinates and altitude. 

80 bands were common in all populations and 

56 were polymorphic (Table 2). 

Except primers UBC 204, UBC 235 and 

UBC 248, all primers generated multiple 

banding patterns with 1 to 8 polymorphic 

amplified DNA bands ranging in size from 250 

to 3000 pb. The polymorphic markers yielded 

44 different electrophoretic banding profiles. 

An example of RAPD banding profile 

generated by the primers UBC 162, UBC 213 

and UBC 225 is presented in Fig.1. Hence, we 

may assume that a large genetic diversity at 

the DNA level characterises the five annual 

caraway populations. 

Estimation of Rp values exhibited a 

collective rate of 30 and varied from 0.4 for 

UBC 211 and UBC 231 primers to 5.2 for 

UBC 264 one with a mean of 2.14 (Table 2). 

UBC 261 and UBC 264 primers were the most 

informative primers as displaying the highest 

Rp rates. Thus, these latter could be referred 

to as the most efficient loci in assessing 

genetic diversity within C. carvi. 

The distinctiveness of the clusters 

identified in the UPGMA derived dendrogram 

exhibited 3 distinct groups (A, B and C) (Fig. 

2). Hence, genetic divergence was dependent 

from the geographic origin and all tested 

populations were differentiated. 

German and Egyptian genotypes housed in 

Group A were referred to as an out-put of 

analysis as diverging significantly from 

Tunisian populations. This has been further 

explained at the DNA level as we were able to 

select a set of RAPD fingerprints unique to 

each of the studied populations (Table 2). For 

example, we counted 5 unique RAPD bands 

for the Tunisian population generated by the 

primers UBC 226, UBC 225, UBC 243, UBC 

261 and UBC 264 and sizing respectively 

700, 835, 1700, 1100 and 975 base pairs. 

40 

Laribi et al. 

The two-dimensional FCA graph allowed 

also significant grouping of the populations 

that were plotted into five sub-plots, 

representing each one population (Fig. 3). 

Population analysis 

Primers varied in their ability to detect 

variation at both within and between 

populations, with among populations H0 ranging from 8.5% (primer UBC-226) to 

100% (primers UBC-162 and UBC-231) 

(Table 2). On average over all primers, the 

population Haouaria (1) exhibited the lowest 

level of within population genetic diversity 

(mean H0 0.012), while the populations 

Menzel Temime (2), Germany (4) and Egypt 

(5) displayed similar estimates of mean H0 (0.022-0.031) (Table 2). However, population 

Souassi (3) was the most variable (mean H0 0.107) (Table 2). 

The results derived from Shannon's index 

showed that variation within (Hpop/Hsp) and 

between (1- Hpop/Hsp) populations was 12.9 

and 87.1%, respectively (Table 2). Thus, high 

level of genetic diversity was detected 

between populations rather than within 

populations. 

A matrix of pair-wise GST values, the 

significance and the effective number of 

migrants (Nm) between populations are 

presented in Table 3. Values of GST ranged 

from 0.288 (between populations Haouaria 

and Menzel Temime) to 0.765 (between 

populations Souassi and Germany), averaging 

0.671. This indicates that all populations may 

be considered different from each other, with 

population Germany (4) being the most 

different from the others and populations 

Haouaria (1) and Menzel Temime (2) being the 

most similar. 

The significant differentiation between 

populations was also revealed in the estimates 

of gene flow (Nm) (Table 3). Values of Nm ranged from a moderate value of 0.465 to 

0.749, averaging 0.585 which indicated low 

gene flow between the five studied 

populations of C. carvi. 



In this study, we fingerprinted a set of five 

annual caraway populations originating from 

Tunisia, Germany and Egypt by means of 

RAPD markers in order to obtain molecular 

data on their genetic background. Thus, we 

have demonstrated the reliability of RAPD 

analysis to detect DNA polymorphisms and 

relationships within C. carvi populations. 

The selected primers were highly 

discriminating since they were characterised 

by relatively high collective Rp rate (30.00), a 

high number of polymorphic markers and 

electrophoretic banding patterns. The primers 

generated 56 polymorphic bands out of 136 

with a mean of 4. Only three primers did not 

allow DNA amplification (Table 2). In RAPD 

literature, the presence of primers does not 

allow amplification to occur (Caetano-Annoles 

1994), whereas others primers yielding faint 

banding profiles (Moreno et al. 1995, Ortiz et 

al. 1997) were reported. In addition, 

according to the results forwarded by Devos 

and Gale (1992), Penner et al. (1993) and 

This et al. (1997), some primers are more 

Laribi et al. 

Table 2. Genetic diversity analysis through RAPD markers and Shannon's index among five annual C. carvi 

populations originated from Tunisia, Germany and Egypt. 

*Population labels are given in Table 1. 

Table 3. Pair-wise GST values (below diagonal) and 

Nm (above diagonal) between the five 

annual C. carvi populations. 

*Population labels are given in Table 1. 

DISCUSSION 

Fig. 1. An example of RAPD banding profile of five 

C. carvi genotypes generated using the 

primers UBC 162, UBC 213 and UBC 225. 

Lane 1: Haouaria, lane 2: Menzel Temime, 

lane 3: Souassi, lane 4: Germany, lane 5: 

Egypt; lane-M: Molecular size marker (1Kb 

DNA ladder). Size of DNA fragments is 

presented on right. 

Fig. 2. UPGMA cluster analysis within the five 

annual C. carvi populations established by 

means of 56 RAPD markers. 

The letters (from A to C) stand for the group 

individualized (1: Haouaria, 2: Menzel Temime, 

3: Souassi, 4: Germany, 5: Egypt). 



Fig. 3. FCA Two-dimensional plot of C. carvi 

populations. (1: Haouaria, 2: Menzel Temime, 

3: Souassi, 4: Germany, 5: Egypt) 

efficient than others in producing stable and 

reproducible profiles. 

The genetic divergence of the populations 

under investigation was confirmed at the DNA 

level. In fact, the UPGMA cluster analysis 

permitted the discrimination of all the 

genotypes and their sorting into 3 main 

groups (Fig. 2). German and Egyptian caraway 

populations (Group A, Fig. 2) diverged 

significantly from Tunisian ones. Thus, 

population clustering was made dependently 

from the original region. In the same way, the 

occurrence of RAPD fingerprints unique to 

each of the studied populations (Table 2) 

suggests that RAPD markers may constitute 

rapid molecular tools for assigning one 

genotype to its origin. On the other hand, 

RAPD markers are suitable to perform genetic 

variation analyses at both intra and interpopulation 

levels (Syankumar and Sasikuma 

2007, Hadian et al. 2008, Zahid et al. 2009). 

According to Shannon's index, high levels 

of genetic diversity were detected between C. 

carvi populations rather than within 

populations (Table 2). The existence of low 

genetic diversity within the species has been 

mostly attributed to self pollination, unless 

other environmental pressures are influencing 

genetic diversity (Archak et al. 2002). In our 

case, despite being an out-crossing and 

insect-pollinated plant (Nemeth et al. 1999, 

Nemeth and Szekely 2000), C. carvi exhibited 

low levels of genetic variation within 

populations. In general, C. carvi 

cenopopulations are grown wild in natural 

habitats such as grasslands and are 

characterized by rich genetic diversity 

42 

Laribi et al. 

strongly influenced by ecological conditions 

(Petraityte 2003, Petraityte and Dastikaite 

2007). Therefore, C. carvi confined to seminatural 

grasslands responds to decreases in 

fragment size by forming smaller populations 

(Kiviniemi 2008). This may have directly 

reduced genetic diversity, and also subjected 

the remaining populations to genetic drift 

(Nunney and Elam 1994). Thus, C. carvi 

population persistence is dependent on 

continuous management that maintains 

habitat quality and creates suitable conditions 

for regeneration, and hence opportunities for 

positive population growth (Kiviniemi 2008). 

Furthermore, another possible explanation 

for the low genetic diversity of C. carvi is the 

influence of human activity. In fact, the recent 

rapid development of tourism and urbanization 

has led to low population densities and 

restricted distribution of C. carvi 

cenopopulations, resulting in loss and 

reduction of genetic diversity. As a result, the 

disappearance of C. carvi cenopopulations 

makes the genetic resources of the species 

poor (Petraityte 2003, Petraityte and 

Dastikaite 2007) and conventional caraway 

breeding is hampered mainly by the limited 

available genetic variation of the caraway 

gene pool (Nemeth 1998). 

Detecting the patterns of variation, in 

terms of polymorphism and differentiation 

among C. carvi populations using RAPD 

markers, is similar to the finding obtained by 

Fu et al. (2003) for Changium smyrnioides 

Wolff (Apiaceae) who also reported that 

variation was be partitioned between rather 

than within populations. However, these 

results are not in accordance with those of 

Barbosa et al. (2010) in a medicinal plant 

Palicourea coriacea (Cham.) K. Schum. 

(Rubiaceae) for which, 23% of genetic 

variability is found among populations and 

77% of genetic variability is within 

populations. 

GST (0.671) and Nm (Nm 0.585, Nm


structure of C. carvi populations. However, 

pollen flow may be inhibited by the distance 

between these populations. Indeed, the 

caraway species have not been able by 

natural means, e.g. by wind or by domestic 

cattle, to exploit all suitable habitat sites 

within grassland fragments (Kiviniemi 2008). 

Furthermore, seed dispersal capacity of this 

plant may be limited because its seeds lack 

morphological adaptations for dispersal. 

Consequently, gene flow among populations 

was expected to be low and genetic 

differentiation to be high. 

The same results were obtained by 

Aegisdóttir et al. (2009) who analyzed 

molecular diversity of a rare alpine plant 

species Campanula thyrsoides L. using five 

polymorphic microsatellite loci. They 

demonstrated that this species showed high 

genetic diversity and considerable population 

differentiation. 

However, these results are in contrast to 

those reported by Sebastien et al. (2010) who 

found low levels of differentiation in a 

medicinal plant Tylophora rotundifolia Buch.- 

Ham. ex Wight using RAPD markers, which 

they attributed to a common genetic pool, 

lack of differential selection pressure, 

diminished mutation rate and short life span of 

the species. 

As expected, the groupings of populations 

were largely in accordance with their 

geographical localities, and the representation 

of genetic relatedness among individuals 

obtained with the two-dimensional graph 

derived from factorial analysis of RAPD 

REFERENCES 

Laribi et al. 

frequency data (Fig. 3) also indicated the low 

level of gene flow among C. carvi populations. 

CONCLUSION 

To conclude, this is the first report on the 

assessment of genetic diversity and 

population differentiation analysis in five 

annual caraway (Carum carvi) populations 

originating from Tunisia, Germany and Egypt, 

using RAPD markers. Though the number of 

populations used in this work does represent 

the total diversity of Tunisian caraway 

populations; nor those of the Egyptian and the 

German populations. We successfully 

provided deep insights on the genetic 

background of the above mentioned 

populations. Considerable genetic diversity 

has been detected within the species. 

Nevertheless, using more informative markers 

such as SSRs and including more populations 

would allow detecting the genetic background 

resources of our local caraway. Therefore, to 

prevent further substantial loss of genetic 

diversity, we should conserve as many 

populations as possible. In this context, we 

suggest that an appropriate strategy for 

sampling may be formulated when ex situ 

conservation is required. Besides, the best in 

situ conservation strategy for C. carvi 

population is to preserve its natural habitats. 

We further aim to include wild germplasm in 

order to select cultivars of economically 

importance and to transfer important genetic 

traits from wild to cultivated species by 

marker-assisted-selection. 

Aegisdottir H, Kuss P, Stöcklin J (2009) Isolated populations of a rare alpine plant show high 

genetic diversity and considerable population differentiation. Annals of Botany 104: 1313- 

1322. 

Archak S, Karihaloo JL, Jain A (2002) RAPD markers reveal narrowing genetic base of Indian 

tomato cultivars. Current Science 82: 1139-1143. 

Barbosa TCS, Sibov ST, Telles MPC, Soares TN (2010) Genetic characterization of natural 

populations of the medicinal plant Palicourea coriacea (Rubiaceae) with molecular markers. 

Genetics and Molecular Research 9: 695-704. 

Bowers JE, Bandmann EB, Meredith CP (1993) DNA fingerprinting and characterization of some 

wine grape cultivars. American Journal of Enology and Viticulture 44: 266-274. 



44 

Laribi et al. 

Caetano-Annoles G (1994) A versatile and universal tool for genome analysis. Plant Molecular 

Biology 25: 1011-1026. 

Dabkevicius Z, Gelvonauskis B, Leistrumaite A (2008) Investigation of genetic resources of 

cultivated plants in Lithuania. Biologija 54: 51-55. 

Devos KM, Gale MD (1992) The use of random amplified polymorphic DNA markers in wheat. 

Theoretical and Applied Genetics 84: 567-572. 

Dolezalova I, Lebeda A, Dziechciarkova M, Kristkova E, Astley D, Wiel C (2003) Relationships 

among morphological characters, isozymes polymorphism and DNA variability-the impact on 

lactuca germplasm taxonomy. Czech Journal of Genetics and Plant Breeding 39: 59-67. 

Eddouks M, Lemhadri A, Michel JB (2004) Caraway and caper: potential anti-hyperglycaemic 

plants in diabetic rats. Journal of Ethnopharmacology 94: 143-148. 

Ene AC, Nwankwo EA, Samdi LM (2007) Alloxan-induced diabetes in rats and the effects of 

black caraway (Carum carvi L.) oil on their body weight. Research Journal of Medicine and 

Medical Sciences 2: 48-52. 

Fracaro F, Zacaria J, Echeverrigaray S (2005) RAPD based genetic relationships between 

populations of three chemotypes of Cunila galioides Benth. Biochemical and Systematics 

Ecology 33: 409-417. 

Fu C, Yingxiong QIU, Kong H (2003) RAPD analysis for genetic diversity in Changium 

smyrnioides (Apiaceae), an endangered plant. Botanical Bulletin of Academia Sinica 44: 13- 

18. 

Gilbert JE, Lewis RV, Wilkinson MJ (1999) Developing an appropriate strategy to assess genetic 

variability in plant germplasm collections. Theoretical and Applied Genetics 98: 1125-1131. 

Hadian J, Tabatabaei SMF, Naghavi MR, Jamzad Z, Ramak-Masoumi T (2008) Genetic diversity 

of Iranian accessions of Satureja hortensis L. based on horticultural traits and RAPD markers. 

Scientia Horticulturae 115: 196-202. 

Hornok L (1986) Effect of environmental factors on growth, yield and on the active principles 

of some spice plants. Acta Horticulturae 188: 69-176. 

Islam MA, Meister A, Schubert V, Kloppstech K, Esch E (2007) Genetic diversity and 

cytogenetic analyses in Curcuma zedoaria (Christm.) Roscoe from Bangladesh. Genetic 

Resources and Crop Evolution 54: 149-156. 

Kamaleeswari M, Kumaraswami Deeptha K, Murugan Sengottuvelan M, Nalini N (2006) Effect 

of dietary caraway (Carum carvi L.) on aberrant crypt foci development, fecal steroids, and 

intestinal alkaline phosphatase activities in 1,2-dimethylhydrazine-induced colon 

carcinogenesis. Toxicology and Applied Pharmacology 214: 290-296. 

Kiviniemi K (2008) Effects of fragment size and isolation on the occurrence of four short-lived 

plants in semi-natural grasslands. Acta Oecologica 33: 56-65. 

Klocke E, Langbehn J, Grewe C, Pank F (2002) DNA fingerprinting by RAPD on Origanum 

majorana. Journal of Herbs, Spices and Medicinal Plants 9: 171-176. 

Lahlou S, Tahraoui A, Zafar Israili Z, Lyoussi B (2007) Diuretic activity of the aqueous extracts 

of Carum carvi and Tanacetum vulgare in normal rats. Journal of Ethnopharmacology 110: 

458-463. 



Laribi et al. 

Lemhadri A, Hajji L, Michel J, Eddouks M (2006) Cholesterol and triglycerides lowering activities 

of caraway fruits in normal and streptozotocin diabetic rats. Journal of Ethnopharmacology 

106: 321-326. 

Lunch M, Milligan BG (1994) Analysis of population genetic structure with RAPD markers. 

Molecular Ecology 3: 91-99. 

McDermott JM, McDonald BA (1993) Gene flow in plant pathosystems. Annual Review of 

Phytopathology 31: 353-373. 

Moreno S, Gogorcena Y, Ortiz JM (1995) The use of RAPD markers for identification of 

cultivated grapevine (Vitis vinifera L.). Scientia Horticulturae 62: 237-243. 

Naderi-Kalali B, Allameh A, Rasaee MJ, Bach HJ, Behechti A, Doods K, Kettrup A, Schramm 

KW (2005) Suppressive effects of caraway (Carum carvi) extracts on 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin-dependent 

gene expression of cytochrome P450 1A1 in the rat H4IIE cells. 

Toxicology in Vitro 19: 373-377. 

Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number 

of individuals. Genetics 89: 583-590. 

Nemeth E (1998) Caraway: The Genus Carum. Harwood Academic Publishers, Amsterdam. 

Nemeth E, Bernath J, Petheo F (1999) Study on flowering dynamic and fertilization properties 

of caraway and fennel. Acta Horticulturae 502: 77-83. 

Nemeth E, Szekely G (2000) Floral biology of medicinal plants. I. Apiaceae species. International 

Journal Horticultural Science 6: 133-136. 

Nunney L, Elam DR (1994) Estimating the effective population size of conserved populations. 

Conservation Biology 8: 175-184. 

Ortiz A, Renaud R, Calzada I, Ritter E (1997) Analysis of Palm cultivars with RAPD markers. 

Journal of Horticultural Science 72: 1-9. 

Papini A (2006) The systematic position of Chamaesciadium C. A. Meyer (Umbelliferae) on the 

basis of nuclear ITS sequence. Flora Mediterranea 16: 5-15. 

Papini A, Banci F, Nardi E (2007) Molecular evidence that genus Carum L. (Apiaceae) is 

polyphyletic. Genetics and Molecular Biology 30(2): 475-482. 

Penner GA, Bush A, Wise R, Kim W, Domier L, Kasha K, Laroche A, Scoles G, Molnar SJ, Fedak 

G (1993) Reproducibility of random amplified polymorphic DNA (RAPD) analysis among 

laboratories. PCR Methods and Applications 2: 341-345. 

Petraityte N (2003) Ex-situ stability of morpho-biochemical properties of common caraway 

(Carum carvi L.). Ekologija (Vilnius) 1: 43-46. 

Petraityte N, Dastikaite A (2007) Agrobiological assessment of wild Carum carvi L. 

cenopopulation biodiversity ex situ. Biologija 53: 74-79. 

Prevost A, Wilkinson MJ (1999) A new system of comparing PCR primers applied to ISSR 

fingerprinting of potato cultivars. Theoretical and Applied Genetics 98:107-112. 

Rafalski JA, Tingey SV (1993) Genetic diagnosis in plant breeding: RAPDs microsatellites and 

machines. Trends in Genetics 9: 275-280. 

Ragot M, Hoisington DA (1993) Molecular markers for plant breeding: comparisons of RFLP and 

RAPD genotyping costs. Theoretical and Applied Genetics 86: 975-984. 



46 

Laribi et al. 

Sebastian VA, D'Cruz L, Subramanian RB, Braganza VJ (2010) Assessment of genetic diversity 

within and among populations of Tylophora rotundifolia using RAPD markers. Gene conserve 

9: 94-117. 

Sneath PHA, Sokal RR (1973) Numerical Taxonomy. The Principle and Practice of Numerical 

Classification. W.H. Freeman and Co., San Francisco. 

Solouki M, Mehdikhani H, Zeinali H, Emamjomeh AA (2008) Study of genetic diversity in 

Chamomile (Matricaria chamomilla) based on morphological traits and molecular markers. 

Scientia Horticulturae 117: 281-287. 

Syamkumar S, Sasikumar B (2007) Molecular marker based genetic diversity analysis of 

Curcuma species from India. Scientia Horticulturae 112: 235-241. 

Tahraoui A, El-Hilaly J, Israili ZH, Lyoussi B (2007) Ethnopharmacological survey of plants used 

in the traditional treatment of hypertension and diabetes in south-eastern Morocco (Errachidia 

province). Journal of Ethnopharmacology 110: 105-117. 

This P, Cuisset C, Boursiquot JM (1997) Development of stable RAPD markers for the 

identification of grapevine rootstocks and the analysis of genetic relationships. American 

Journal of Enology and Viticulture 48: 492-501. 

Toxopeus H, Bouwmeester HJ (1993) Improvement of caraway essential oil and carvone 

production in The Netherlands. Industrial Crops and Products 1: 295-301. 

Williams JGK, Kubelik AR, Livak KJ, Rafalski JA (1990) DNA polymorphisms amplified as 

arbitrary primers are useful genetic markers. Nucleic Acids Research 18: 6531-6535. 

Wright S (1951) The genetical structure of populations. Annals of Eugenics 15: 323-354. 

Yeh FC, Chong DKX, Yang RC (1995) RAPD variation within and among natural populations of 

trembling aspen (Populus tremuloides) from Alberta. Journal of Heredity 86: 454-460. 

Yeh FC, Yang R, Boyle T (1999) POPGENE Version 1.32. Microsoft Window-Based Freeware 

for Population Genetic Analysis. University of Alberta, Edmonton. 

Zahid NY, Abbasi NA, Hafiz IA, Ahmad Z (2009) Genetic diversity of indigenous fennel 

(Foeniculum vulgare Mill.) germoplasm in Pakistan assessed by RAPD markers. Pakistan 

Journal of Botany 41: 1759-1767. 

Zoghlami N, Mliki A, Ghorbel A (2001) Evaluation of genetic diversity among Tunisian 

grapevines by RAPD markers. Vitis 40: 31-37. 

Zoghlami N, Chrita I, Bouamama B, Gargouri M, Zemni H, Ghorbel A, Mliki A (2007) Molecular 

based assessment of genetic diversity within Barbary fig (Opuntia ficus indica (L.) Mill.) in 

Tunisia. Scientia Horticulturae 113: 134-141. 



Laribi et al. 

Frenk Kimyonu (Carum carvi) Popülasyonlarinda Genetik Çesitliligin RAPD Temelli Incelenmesi 

Özet 

Tunus, Almanya ve Misir kaynakli yillik Frenk kimyonuna (Carum carvi) ait bes popülasyon 

arasindaki genetik çesitlilik ve farklilasma, ilk kez incelendi. Genetik çesitlilik, popülasyon yapisi ve 

gen akisi ile alakali Random Amplified Polymorphic DNA (RAPD) markör verileri elde edildi ve 

incelendi. Analiz edilen popülasyonlarda; 14 primer, toplam 136 ayird edilebilir ve tekrarlanabilir 

bant üretti. Bunlardan 56 tanesi polimorfik idi. UPGMA yigin analizi bütün genotipin ayirt 

edilmesini ve 3 ana grupta toplanmasini sagladi. Tunus kimyon popülasyonlari, anlamli ölçüde 

Alman ve Misir popülasyonlarindan ayrildi. Popülasyon gruplanmasi, cografik kaynaktan bagimsiz 

olarak yapildi. Çalisilan popülasyonlara özel RAPD izleri seçebildigimiz için, DNA seviyesinde de bu 

durumu göstermis olduk. Ek olarak, RAPD frekans verilerinin analizinden elde edilen boyutsal 

grafik; genotipleri, her biri popülasyonu temsil eden bes alt dal halinde gruplandirmamizi sagladi. 

Shannon indeks degerleri; farkliligin popülasyonlarin kendi içinde degil, popülasyonlar arasinda 

gerçeklestigini gösterdi. Bu sonuçlar, C. carvi populasyonlari arasinda önemli genetik farkliliklar 

oldugunu göstermistir. 

Anahtar Kelimeler: Carum, çesitlilik, farklilik, popülasyon.

RAPD-based assessment of genetic diversity among annual ...

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?