Relationships of Vigna unguiculata (L.) Walp., V. vexillata (L.) A. Rich. and species of section Vigna based on isozyme variation

Summary Isozyme variation in 25 accessions of wild and cultivated Vigna unguiculata, 49 accessions of seven wild species belonging to section Vigna, and 11 accessions of V. vexillata (subgenus Plectrotropis) was scored at 17 putative loci to assess genetic relationships within and among species. The wild species selected for this study are among those which carry important agronomical traits useful in cowpea (V. unguiculata) breeding programs. Allelic frequencies were calculated and Nei's genetic distances were obtained. Low levels of intraspecific variation were observed for V. heterophylla, V. luteola and V. racemosa, whereas the other species showed a higher polymorphism. Vigna unguiculata possessed intraspecific genetic distances comparable to those previously found by other authors. Most of the isozyme variation was apportioned among species. Although V. luteola and V. marina had an interspecific genetic distance resembling the range observed at intraspecific level, all the other species showed very high interspecific distances. Vigna unguiculata was relatively closer genetically to V. vexillata than to the species belonging to section Vigna.Abbreviations AUS Australia - BDI Burundi - BRA Brazil - BWA Botswana - CAF Central African Republic - GHA Ghana - CMR Cameroon - COG Congo - RI Costa Rica - EGY Egypt - ETE Ethiopia - GAB Gabon - GRC Greece - ITA Italy - KEN Kenya - MOZ Mozambique - NER Niger - NGA Nigeria - PAN Panama - RWA Rwanda - TCD Chad - TZA Tanzania - ZAF South Africa - ZAR Zaire - ZMB Zambia     

Genetic Diversity of the Vigna Germplasm from Thailand and Neighboring Regions Revealed by AFLP Analysis

Thailand is a center of diversity for section Angulares of the Asian Vigna (genus Vigna subgenus Ceratotropis) and 4 Vigna species are cultivated in Thailand. Using newly collected wild and cultivated germplasm of Vigna from Thailand and outgroup accessions AFLP analysis was conducted to clarify genetic diversity and relationships. The results suggest that cultivated V. umbellata and V. mungo evolved from wild relatives in a single domestication event. Vigna umbellata is poorly differentiated from its wild and weedy relatives compared to V. mungo. Results suggest northern Thailand and the neighboring Shan state, Myanmar, is the probable center of domestication for V. umbellata as wild accessions from this area and cultivated rice bean from a wide area in Asia are not greatly diverged. Vigna minima, V. tenuicaulis and V. exilis accessions in Thailand are well differentiated with considerable intra-specific variation. Vigna hirtella consists of two well differentiated subgroups, suggesting taxonomic revision may be necessary. Close genetic relationships between V. radiata and V. grandiflora, and between V. mungo and V. trinervia are confirmed. Naturally growing V. mungo populations in northern Thailand appear to be true wild species as they are well differentiated from Indian wild and Thai cultivated populations. The origin of naturally growing cowpea in Thailand needs to be further studied using a more comprehensive set of materials. This study clarifies inter and intra-specific genetic diversity and inter species relationships of Thai Vigna species.     

Genetic diversity in cowpea [Vigna unguiculata (L.) Walp.] as revealed by RAPD markers

The present study, using RAPD analysis, was undertaken to characterize genetic variation in domesticated cowpea and its wild progenitor, as well as their relationships. The materials used consisted of 26 domesticated accessions, including accessions from each of the five cultivar-group, and 30 wild/weedy accessions, including accessions from West, East and southern Africa. A total of 28 primers generated 202 RAPD bands. One hundred and eight bands were polymorphic among the domesticated compared to 181 among wild/weedy cowpea accessions. Wild accessions were more diverse in East Africa, which is the likely area of origin of V. unguiculata var. spontanea. Var. spontanea is supposed to have spread westward and southward, with a loss of variability, loss counterbalanceed in southern Africa by introgressions with local perennial subspecies. Although the variabilty of domesticated cowpea was the highest ever recorded, cultivar-groups were poorly resolved, and several results obtained with isozyme data were not confirmed here. However primitive cultivars were more diverse than evolved cultivars, which still suggests two consecutive bottlenecks within domesticated cowpea evolution. As isozymes and AFLP markers, although with a larger number of markers, RAPD data confirmed the single domestication hypothesis, the gap between wild and domesticated cowpea, and the widespread introgression phenomena between wild and domesticated cowpea.     

Determining genetic diversity between lines of <Emphasis Type="Italic">Vigna unguiculata</Emphasis> subspecies by AFLP and SSR markers

AFLP (Amplified Fragment Length Polymorphisms) and SSR (Simple Sequence Repeat) markers were utilized to assess genetic diversity and relatedness between Vigna unguiculata subspecies. Three AFLP primer combinations and 10 SSR primer sets successfully identified closely related accessions, and the presence of heterogeneity in some accessions. AFLP methodology was successful in separating different species of Vigna. However, the level of intra-subspecies variation was as great as was the interspecies variation with both marker methods. The number of markers employed was insufficient to successfully group the subspecies into distinct clades.     

Assessment of genetic diversity in cultivars and wild accessions of Luohanguo (<Emphasis Type="Italic">Siraitia grosvenorii</Emphasis> [Swingle] A. M. Lu et Z. Y. Zhang), a species with edible and medicinal sweet fruits endemic to southern China,using RAPD and AFLP markers

Genetic variation of wild populations and cultivars of Luohanguo (Siraitia grosvenorii), a plant species endemic to southern China, was assessed using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers. Based on the results for 130 individuals from seven populations, a high level of genetic diversity of Luohanguo was observed at the species level. The percentage of polymorphic loci (P) was 89.4%, Nei’s gene diversity (H _e) was 0.239, and Shannon’s information index (H _o) was 0.373 based on the combined AFLP and RAPD data. There was a high degree of genetic differentiation, with 45.1% of the genetic variation attributed to differences between the populations. The genetic diversity of the Luohanguo cultivars is much lower than that of wild populations (P = 41.8%, H _e = 0.141, H _o = 0.211), and a distinct genetic differentiation is observed between the cultivars and wild accessions. The pool of genetic variation in the wild populations provides an excellent gene resource for Luohanguo breeding.     

A chemosystematic study of the flavonoids of Vigna

Summary Flavonoid chemical markers together with other biochemical markers and/or morphological data can provide useful characters for defining species in Vigna genus. In this paper flavonid HPLC fingerprints have been utilized to assess taxonomic relationships amongst species and accessions of Vigna. Within the taxa analysed there is evidence of both infra- and interspecific chemical variations. In some species chromatographic data support evidence for the existence of different flavonoid chemotypes, which may reflect differences in geographic origin. As far as interspecific relationships are concerned, species belonging to sections Vigna, Plectotropis and Ceratotropis do not show flavonoid glycosides in common with cultivated lines of Vigna. By contrast, some relationships have been found between cultivated lines and wild species of section Catiang. Kaempferol-3-rutinoside and an unidentified kaempferol-glycoside were the only flavonoids detected in the three subgenera analysed. Finally, a greater variability in flavonoid aglycone class and glycosylation pattern has been observed in cultivars of V. unguiculata (L.) Walp. compared to the wild species. This observation may confirm that cultivation and/or domestication may cause or increase species diversification.     

Analysis of population substructure,genetic differentiation and phylogenetic relationships among selected Asiatic <Emphasis Type="Italic">Vigna</Emphasis> Savi species

Random amplified polymorphic DNA markers were used to study sub-structure and genetic differentiation amongst 31 populations (seven cultivated and 24 wild populations) belonging to 14 Asiatic Vigna species. Ten pre-selected RAPD primers generated 152 polymorphic amplification products. Estimates of polymorphism indices were higher for the wild taxa in comparison to the cultivated forms. F_ST values between populations ranged from 0.111 to 0.801 and Nei’s genetic diversity values between and within species varied from 0.26 to 0.70 and 0.04 to 0.56 respectively. The high F_ST and F_CT values indicated strong subdivision of populations and high differentiation among species. Analysis of molecular variance was performed by grouping the populations conforming to specific species. AMOVA was also performed separately to better resolve the differentiation of species within mungo–radiata complex. Molecular phylogenetic relationships amongst the species of radiata–mungo complex; namely, black gram (V. mungo (L.) Hepper), green gram (V. radiata (L.) Wilczek), V. radiata var. sublobata, V. radiata var. setulosa, V. mungo var. silvestris and V. hainiana, were studied through cluster analyses. Two distinct groups were recognized within the complex, with population samples of V. hainiana forming one cluster. Further, V. hainiana appeared to be equidistant to both V. radiata and V. mungo.     

Genetic relationships among cultivated and wild <Emphasis Type="Italic">Vigna angularis</Emphasis> (Willd.) Ohwi et Ohashi and relatives from Korea based on AFLP markers

Genetic variation and relationships among members of the azuki bean complex (Vigna angularis) including wild (V. angularis var. nipponensis), weedy, and cultivated types (V. angularis var. angularis), V. nakashimae, and rice bean (V. umbellata) from Korea were examined using the Amplified fragment length polymorphism (AFLP) method. AFLP analysis of 50 accessions revealed 333 (72.1%) polymorphic fragments out of 462 fragments amplified using seven primer combinations. The number of polymorphic fragments within each species was 70 in the azuki bean complex and 41 in V. nakashimae, but there was no polymorphism in rice bean. The number of shared fragments among species ranges from 142 between the azuki bean complex and V. nakashimae to 166 between the azuki bean complex and rice bean. Within the azuki bean complex, the range of shared bands was from 231 between cultivated and weedy types to 238 between cultivated and wild types. A dendrogram generated from Jaccard’s similarity matrix was divided into three groups, which correspond to V. nakashimae, azuki bean complex, and rice bean. The relationship between azuki bean and rice bean is closer than between azuki bean and V. nakashimae. Phenetic distances averaged 0.502 between the azuki bean complex and V. nakashimae and 0.467 between the azuki bean complex and rice bean. Within the azuki bean complex, the weedy type was more closely related to wild than cultivated types. But UPGMA dendrogram of the azuki bean complex reveals that each type is not clearly isolated. These results will help to understand genetic diversity and evolutionary dynamics of Vigna in Korea.     

RAPD polymorphisms in Aegilops geniculata Roth (Ae. ovata auct. non L.)

Genetic diversity of eighteen accessions of Ae. geniculata (2n=28; UUMM) was assessed using the random amplified polymorphic DNA (RAPD) technique. We optimized RAPD conditions including the template DNA, the concentration of AmpliTaq DNA polymerase Stoffel fragment, and MgCl₂ concentration for revealing polymorphisms. Thirty-eight decamer oligonucleotides were individually used as primers under optimized conditions. Seventeen of these primers produced polymorphic RAPDs among the 18 accessions of Ae. geniculata. Polymorphisms were recorded by noting presence or absence of an amplification product from the total genomic DNA. Comparisons of unique and shared amplification products of each pair of accessions were used to generate genetic similarity coefficients (GSCs). These GSCs were used to construct a phenogram using an unweighted pair-group method with arithmetical averages (UPGMA). The phenogram shows that RAPD data is useful in the measurement of genetic variation or similarity within a species. It also indicates that we can select eight or nine accessions of the eighteen accessions to maintain at least 80% genetic variability of the Chinese collection of Ae. geniculata. Address for correspondence: Dr X-Y. Zhang, USDA-ARS-FRRL, Utah State University, Logan, UT 84322-6300, who is visiting the USA under an agreement between USDA-ARS and CMA-CAAS on germplasm resources.     

Comparative analysis of genetic diversity using molecular and morphometric markers in Andrographis paniculata (Burm. f.) Nees

Andrographis paniculata is a medicinal plant of immense therapeutic value. The present study was aimed to elucidate its genetic diversity based on morphochemical and RAPD markers from 53 accessions belonging to 5 eco-geographic regions. Analysis of variance and D ² statistics revealed significant differences in all the metric traits and sufficient inter-cluster distances indicating considerable diversity among the accessions. The complementary approach of RAPD was used to evaluate the genetic dissimilarities among all the accessions using 6 highly polymorphic primers. The average proportion of polymorphic loci across primers was 96.28%. The molecular genetic diversity based on Shannon index per primer averaged 5.585 with values ranging from 3.08 to 8.70 indicating towards wide genetic base. RAPD based UPGMA and D ² cluster analysis also revealed that various accessions available in different eco-geographic regions might have originated from native places of wild abundance. Similarity matrices were generated for molecular markers and morphometric data to determine the degree of congruence between the two. A highly significant but low correlation (r = 0.547, P < 0.001) was obtained thus implying the correspondence between the two. The species is hermaphroditic and a habitual inbreeder. The present study yielded a typical triangular congruence between its breeding system, morphometric traits and RAPD markers thus elucidating the usefulness of complementary approaches to make diversity analysis more explanatory and purposeful for optimum genetic amelioration and effective conservation of its genotypic variability.     

Genetic diversity among varieties of Chia (Salvia hispanica L.)

Chia, Salvia hispanica L., was a staple crop in pre-Columbian Mesoamerica. Despite the great potential of the species as an oilseed crop, little research related to domesticated and wild varieties exists. A study was undertaken to assess genetic diversity among 38 wild and domesticated accessions of S. hispanica collected throughout Mesoamerica by using RAPD markers. Genetic diversity was higher among wild varieties (H _G= 0.15) than all domesticated varieties (H _G= 0.10) and modern commercial domesticated varieties (H _G= 0.02), suggesting a slight loss of diversity accompanying domestication and a near lack of diversity in modern commercial varieties. In addition, the preliminary results indicate that the center of genetic diversity is in the highlands of western Mexico.     

Assessing Genetic Diversity of Chinese Cultivated Barley by STS Markers

To assess the genetic diversity among China’s cultivated barley, sequence tagged site (STS) marker analysis was carried out to characterize 109 morphologically distinctive accessions originating from five Chinese eco-geographical zones. Fourteen polymorphic STS markers representing at least one in each chromosome were chosen for the analysis. The 14 STS markers revealed a total of 47 alleles, with an average of 3.36 alleles per locus (range 2–8). The proportion of polymorphic loci per population averaged 0.84 (range 0.71–1.00); the mean gene diversity averaged 0.39 (range 0.28–0.49). The means of P and H_e were highest in the Yangtze reaches and Southern zone (P = 1.00; H_e = 0.46) and lowest (P = 0.71 He = 0.28) in the Yellow river reaches zone. The STS diversity in different zones is quite different from the morphology diversity. The STS variation was partitioned into 17% among the zone and 83% within the zone. Both cluster and principal coordinate analyses clearly separated the accessions into a dispersed group (mostly two-rowed barley with a lower mean GS value) and a concentrated group (mostly six-rowed barley with a higher mean GS value) according to the spike characteristic with only a few exceptions. The accessions from the Qinghai-Tibet plateau formed a distinctive subgroup and can be distinguished from the concentrated group. The role of Tibet in the origin and evolution of cultivated barley has been discussed.     

Preliminary study of genetic diversity in Swedish flax (Linum usitatissimum)

To investigate the genetic diversity of Linum usitatissimumL. in Sweden, 18 accessions, including 13 cultivars and five landraces, were analysed. This study was based on genetic variation in three enzyme systems (i.e., PGD, GPI and MDH) by using horizontal starch gel electrophoresis. The total genetic diversity of the studied flax material was very high (H _T= 0.62). Even though the highest genetic diversity lies within the accessions (G _ST= 0.07), a clear differentiation between fibre and oil flax was found with respect to three polymorphic loci (Pgd-1, Gpi-2 and Mdh-1). A phenogram, based on Nei's genetic distances between the accessions studied, showed five clearly defined groups but with low variation within the groups. The unexpected high genetic diversity found within accessions in the studied flax material may indicate that flax is more outbreeding than earlier believed.     

Analysis of genetic diversity in Turkish sesame (Sesamum indicum L.) populations using RAPD markers⋆

The genetic diversity of 38 cultivated populations of Sesamum indicum L. from four different regions of Turkey was estimated at the DNA level with the random amplified polymorphic DNA (RAPD) technique. Sixty-one bands were obtained for all populations 78% of which were polymorphic. Analysis of molecular variance (AMOVA) was used to investigate the genetic diversity of the populations which yielded highly significant differences among populations within regions (91.9% of the total genetic diversity). According to AMOVA and Shannon's index that were performed separately for each region, the highest value of genetic variation was observed among Northwest region populations (CV = 7.7; H₀ = 0.304) and lowest in the Southeast regions' populations (CV = 2.6; H₀ = 0.068). Nei and Li's similarity index was calculated and phylogenetic tree was established using the neighbor-joining algorithm. This phenetic analysis grouped 35 of 38 accessions in six groups leaving three highly diverse accessions outside. Wagner phylogenetic method was used to assess the phylogenetic relationships among the populations. In the majority-rule consensus tree, only 7 of the 32 forks showed above 60% occurrence. Using Principal Coordinate Analysis (PCO) of the RAPD data set, the groups were clearly separated along the first three axis. These results indicate that RAPD technique is useful for sesame systematics, and should be valuable for the maintenance of germplasm banks and the efficient choice of parents in breeding programs.     

Using isozyme polymorphism to assess genetic variation within cultivated yams (Dioscorea cayenensis/Dioscorea rotundata complex) of the Republic of Benin

Four hundred and sixty-seven accessions of cultivated Guinea yam (Dioscorea cayenensis/Dioscorea rotundatacomplex) collected from different localities of Benin Republic were analysed to study isoenzymatic variability in seven enzyme systems: aspartate aminotransferase (AAT), esterase (EST), glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), phosphoglucoisomerase (PGI), and shikimate dehydrogenase (SKDH) using starch gel electrophoresis. Polymorphism was observed in all of the enzyme systems and a total of 62 electromorphs of different frequency and variability patterns were recorded. Different combinations of banding patterns of these systems led to identification of 227 different cultivars within the 467 accessions analysed. For an old and vegetatively propagated crop (with a considerable number of vernacular names) such as yam, and for which a high rate of duplication is expected, the 227 cultivars were found to be good enough to be considered as the adequate number of accessions representing the diversity in the germplasm analysed. Cluster analysis (UPGMA) produced a most likely division of the 467 accessions into two groups corresponding to D. rotundata Poir. and D. cayenensis Lam., supporting the concept that the two forms of guinea yam represent different genetic entities. The different clusters formed within the white yams (D. rotundata) did not exactly conform to the known cultivar groups. Additional polymorphic enzymes are needed for an accurate isozyme-based genetic discrimination of most of the cultivar groups.     

Genetic diversity of cowpea [<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.] in four West African and USA breeding programs as determined by AFLP analysis

Cowpea is an important grain legume and hay crop of many tropical and subtropical regions, especially in the dry savanna region of West Africa. The cowpea gene pool may be narrow because of a genetic bottleneck during domestication. Genetic variation within specific breeding programs may be further restricted due to breeding methods, ‘founder effects’ and limited exchange of germplasm between breeding programs. Genetic relationships among 60 advanced breeding lines from six breeding programs in West Africa and USA, and 27 landrace accessions from Africa, Asia, and South America were examined using amplified fragment length polymorphism (AFLP) markers with six near infrared fluorescence labeled EcoRI + 3/1bases/MseI + 3/1bases primer sets. A total of 382 bands were scored among the accessions with 207 polymorphic bands (54.2%). Despite a diverse origin, the 87 cowpea accessions shared a minimum 86% genetic similarity. Principal coordinates analysis showed clustering of breeding lines by program origin, indicating lack of genetic diversity compared to potential diversity. Accessions from Asia and the Americas overlapped and were distinct from West African breeding lines, indicating that germplasm from Asia and the Americas have common origins outside West Africa. US and Asian breeding programs could increase genetic variability in their programs substantially by incorporating germplasm from West Africa, while national programs in West Africa should consider introgression of Asian germplasm and germplasm from other parts of Africa into their programs to ensure long-term gains from selection.     

Landraces of cowpea, <Emphasis Type="Italic">Vigna</Emphasis><Emphasis Type="Italic">unguiculata</Emphasis> (L.) Walp., as potential sources of genes for unique characters in breeding

Cowpea landraces belonging to Vigna unguiculata (L.) Walp. subsp. unguiculata cv.-gr. unguiculata and cv.-gr. sesquipedalis collected from part of the Deccan Plateau and West Coast of India were evaluated to (i) identify the diverse source(s) of variation for improved characters like pods/peduncle and seed index (ii) study the response of landraces for adaptation to drought and heat stress and (iii) understand the breeding value of a landrace in the genetic improvement of a popular cowpea cultivar. Landraces were evaluated for various morphological characters, pods/peduncle, seed index and other economically important agronomic traits, rust resistance and drought and heat tolerance in different years and environments. Landraces were found as an important source of genetic variability for pods/plant, pods/peduncle, better pod filling ability (seed index), grain yield/plant as well as drought and heat tolerance and rust resistance. Hybridization between C 152 (cv.) and DWDCC 016 (landrace) resulted in release of new variation not present in the two parents. Thus the landrace, DWDCC 016, can be utilized to improve cultivated varieties by transferring to them the economically valuable traits like pods/peduncle and seed index thereby enhancing realisation of sink potential and ultimately grain yield in a sustainable way.     

Population diversity of the Vicia sativa agg. (Fabaceae) in the flora of the former USSR deduced from RAPD and seed protein analyses

The genetic diversity of 58 wild and weedy populations representing taxa within the V. sativa aggregate from the former USSR, 4 cultivars of V. sativa, 2 accessions of V. cordata and 3 accessions of V. macrocarpa from Mediterranean countries were analysed using randomly amplified DNA fragments (RAPDs) and seed protein electrophoresis (SDS-PAGE). Interspecific variation between taxa in the V. sativa aggregate could readily be detected using both techniques. RAPDs and seed protein patterns were found to be an effective means of identifying accessions that cannot be identified clearly by morphological criteria alone. RAPD and seed protein analysis revealed a clear relationship between observed genetic variation of populations and their geographical distribution. Populations from each region had their own gene pools. Geographical variation was detected in V. segetalis. The degree of genetic divergence between local populations was usually related to proximity. In several locations where wild and weedy populations of different V. sativa agg. taxa grow sympatrically, intermediate forms could be detected at the DNA and protein levels. Both RAPD and seed protein analysis support the view that the V. sativa aggregate consists of 8 taxa warranting recognition at the species level. Several species in this aggregate are evolving intra-specific groups which can readily be detected at the molecular level.     

Study of allozyme diversity in guar [Cyamopsis tetragonoloba (L.) Taub.] germplasm

Guar [Cyamopsis tetragonoloba (L.) Taub.] is an important commercial crop of India, mainly cultivated in the northwestern part and neighboring areas of Pakistan. High morphological diversity is reported in guar. The present study attempts to analyze the genetic diversity in cultivated guar using allozyme polymorphism and compare it with reported morphological diversity. Accessions for the study were selected from a set of guar germplasm conserved at the National Bureau of Plant Genetic Resources, New Delhi. The morphological data of 108 selected accessions was subjected to UPGMA clustering. Fifty-five accessions were then picked randomly from the clusters generated. Tris-borate system (pH 8.3) was found to be most effective for detection of allozyme polymorphism in guar. Ten enzymes were selected for the study out of 18 systems tried, namely, malate dehydrogenase, malic enzyme, phosphogluco isomerase, glutamate oxaloacetate transaminase, esterase, peroxidase, alcohol dehydrogenase, glutamate dehydrogenase, glucose 6-phosphate dehydrogenase and shikimate dehydrogenase. A total of 20 loci were detected with 34 alleles. The number of polymorphic loci in different accessions was observed to be highly variable (0–9). An average of 1.27 allozyme per polymorphic locus was observed. The allele frequency data of the 55 accessions was used to determine genetic diversity parameters. The dendrogram generated on genetic distance showed three clusters in the distance range of 0–0.28. Most of the accessions were in the first clusters in a narrow distance range of 0–0.08. The second cluster consisted of improved varieties and accession from Pakistan. The third cluster of accessions was genetically more distant but there was no definite segregation of accessions according to the place of collection. The accessions from south west Rajasthan were scattered in all the three clusters showing higher variability in this region, also supported by higher morphological variation. The genetic distance was low among accessions of guar, indicating low diversity in India.     

Genetic diversity in Tunisian perennial ryegrass revealed by ISSR markers

The genetic diversity in Tunisian perennial ryegrass (Lolium perenne) was examined by the help of inter-simple sequence repeats (ISSR). Starting from eighteen accessions, a large number of polymorphic ISSR markers were currently generated using appropriate primers (a total of 136, which average of 12.6 polymorphic bands/primer). These markers were considered to estimate the genetic distance among accessions and to draw phylogenetic trees. Our data provide evidence of a high degree of genetic diversity in Tunisian ryegrass. In addition, both cultivars and wild types present a high degree of divergence suggesting a complex domestication process in this crop. Moreover, spontaneous populations of Tunisian ryegrass have been identified as important ecotypes that are suitable in selection programs to improve grasslands.