Identification of <Emphasis Type="Italic">Aegilops</Emphasis> L. species and <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. based on chloroplast DNA

The genus Aegilops L. is a very important genetic resource for the breeding of bread wheat Triticum aestivum. Therefore, an accurate and easy identification of Aegilops species is required. Traditionally, identification of Aegilops species has relied heavily on morphological characters. These characters, however, are either not variable enough among Aegilops species or too plastic to be used for identification at the species level. Molecular markers that are more stable within species, therefore, could be the alternative strategy towards an accurate identification. Since the chloroplast DNA has a lower level of evolution compared to the nuclear genome, an attempt was made in this study to investigate polymorphism in the chloroplast DNA among 21 Aegilops species (including Ae. mutica that is now known as Amblyopyrum muticum) and between the latter and T. aestivum to generate markers for the diagnosis of all targeted species. Cleaved amplified polymorphic sequence (CAPS) applied on 22 coding and non-coding chloroplast regions using 80 endonucleases and sequencing of two of those regions revealed little polymorphism between T. aestivum and the various Aegilops species examined and to a less extent was the variation among Aegilops species. Polymorphism observed among species analysed allowed the discrimination of T. aestivum and 12 Aegilops species.     

Red List assessment of nine <Emphasis Type="Italic">Aegilops</Emphasis> species in Armenia

The aims of this study are to determine the geographical and ecological distribution of nine Aegilops species in Republic of Armenia and to make an assessment of their IUCN Red List status, using the IUCN Red list categories and criteria, in order to develop an in situ conservation strategy for wild relatives of wheat in Armenia. Ecogeographic surveys of nine Aegilops species were undertaken over 2 years in Armenia. They included a herbarium survey followed by extensive ground-truthing field surveys where targeted Aegilops species occur. The study showed that of the nine Aegilops species studied, four are threatened and of these, Ae. mutica and Ae. crassa are critically endangered. The latter species may even be extinct in Armenia. Ae. neglecta and A. biuncialis are endangered. Additional studies are required to assess the threat status of Ae. umbellulata. Ae. columnaris was assessed as near threatened, while the remaining species (Ae. triuncialis, Ae. cylindrica and Ae. tauschii) are of least concern. There has been a dramatic decline in the genetic resources of Aegilops species during recent years in Armenia as a result of adverse human impacts such as expansion of agriculture, urbanization and uncontrolled grazing. Several species, especially Ae. mutica and Ae. crassa, should be prioritized in conservation activities in Armenia. Efforts should be made to conserve genetic diversity of crop wild relative species both in situ and ex situ, bearing in mind that their germplasm carries potentially valuable information (traits) that can improve adaptability and productivity of cultivated wheat varieties.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

A semi-fertile interspecific hybrid of <Emphasis Type="Italic">Brassica rapa</Emphasis> and <Emphasis Type="Italic">B. nigra</Emphasis> and the cytogenetic analysis of its progeny

Black rot is a bacterial disease of Brassica rapa caused by Xanthomonas campestris pv. campestris (Xcc.). Sources of resistance to this disease within B. rapa are insufficient and control measures are limited, making the development of resistant breeding lines extremely important. Certain lines of B. nigra exhibit very high resistance to Xcc. For this study, an interspecific cross between Brassica rapa and B. nigra was performed, and a total of 6 F₁ hybrids were obtained through ovary culture. Five plants (H_1–5) were relatively slow-growing, entirely sterile, and had 18 chromosomes in the majority of pollen mother cells (PMCs). GISH analysis showed that most of the PMCs had 8 B. nigra chromosomes, which indicated the expected AB genomic constitution. The last plant (H₆) was partially fertile and the majority of PMCs contained 10 chromosomes of B. rapa and 16 chromosomes of B. nigra, indicating an ABB genomic constitution. Ovary culture techniques were not necessary for the development of the first- or second-backcross generation. Sequence-related amplified polymorphism analysis of F₁, BC₁, and BC₂ plants indicated that some fragments from B. nigra were lost, particularly in the genome of BC₁/BC₂ in successive generation(s). The BC₂ plants expressing good resistance to Xcc. were observed.     

Transfer of leaf rust and stripe rust resistance from <Emphasis Type="Italic">Aegilops umbellulata</Emphasis> Zhuk. to bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Aegilops umbellulata acc. 3732, an excellent source of resistance to major wheat diseases, was used for transferring leaf rust and stripe rust resistance to cultivated wheat. An amphiploid between Ae. umbellulata acc. 3732 and Triticum durum cv. WH890 was crossed with cv. Chinese Spring Ph ^I to induce homoeologous pairing between Ae. umbellulata and wheat chromosomes. The F₁ was crossed to the susceptible Triticum aestivum cv. ‘WL711’ and leaf rust and stripe rust resistant plants were selected among the backcross progenies. Homozygous lines were selected and screened against six Puccinia triticina and four Puccinia striiformis f. sp. tritici pathotypes at the seedling stage and a mixture of prevalent pathotypes of both rust pathogens at the adult plant stage. Genomic in situ hybridization in some of the selected introgression lines detected two lines with complete Ae. umbellulata chromosomes. Depending on the rust reactions and allelism tests, the introgression lines could be classified into two groups, comprising of lines with seedling leaf rust resistance gene Lr9 and with new seedling leaf rust and stripe rust resistance genes. Inheritance studies detected an additional adult plant leaf rust resistance gene in one of the introgression lines. A minimum of three putatively new genes—two for leaf rust resistance (LrU1 and LrU2) and one for stripe rust resistance (YrU1) have been introgressed into wheat from Ae. umbellulata. Two lines with no apparent linkage drag have been identified. These lines could serve as sources of resistance to leaf rust and stripe rust in breeding programs.     

Production and cytogenetics of trigeneric hybrid involving <Emphasis Type="Italic">Triticum</Emphasis>, <Emphasis Type="Italic">Psathyrostachys</Emphasis> and <Emphasis Type="Italic">Secale</Emphasis>

Trigeneric hybrids may help establish evolutionary relationships among different genomes present in the same cellular-genetic background, and also offers the possibility to transfer different alien characters into cultivated wheat. In this study, a new trigeneric hybrid involving species from the Triticum, Psathyrostachys and Secale was synthesized by crossing wheat-P. huashanica amphiploid (PHW-SA) with wheat-S. cereale amphiploid (Zhongsi 828). The crossability of F₁ hybrid was high with 35.13%, and the fertility was 41.95%. The morphological characteristics of F₁ plants resembled the parent Zhongsi 828. The trigeneric hybrids pollen mother cells (PMCs) regularly revealed averagely 19.88 univalents, 9.63 ring bivalents, 3.97 rod bivalents, 0.60 trivalents and 0.03 tetravalents per cell. Multivalents consisted of trivalents and tetravalents can be observed in 52.7% of cells. A variation of abnormal lagging chromosome, micronuclei and chromosome bridge were formed at anaphase I and telophase II. The mean chromosomes number of F₂ progenies was 2n = 46.13, and the distribution range was 42–53. GISH results revealed that most F₂ plants had 6–12 S. cereale chromosomes, and only 0–2 P. huashanica chromosomes were detected. The results indicated that S. cereale chromosomes can be preferentially transmitted in the F₂ progenies of trigeneric hybrid than P. huashanica chromosomes. A survey of disease resistances revealed that the stripe rust resistance from the PHW-SA were completely expressed in the F₁ and some F₂ plants. The trigeneric hybrid could be a useful bridge for the transference of P. huashanica and S. cereale chromatins to common wheat.     

Characterization of fertile amphidiploid between <Emphasis Type="Italic">Raphanus sativus</Emphasis> and <Emphasis Type="Italic">Brassica alboglabra</Emphasis> and the crossability with <Emphasis Type="Italic">Brassica</Emphasis> species

A fertile amphidiploid × Brassicoraphanus (RRCC, 2n = 36) between Raphanus sativus cv. HQ-04 (2n = 18, RR) and Brassica alboglabra Bailey (2n = 18, CC) was synthesized and successive selections for seed fertility were made from F₄ to F₁₀. F₁₀ plants exhibited good fertility with 14.9 seeds per siliqua and 32.3 g seeds per plant. Cytological observation revealed that frequent secondary pairing occurred among 3 chromosome pairs in pollen mother cells of plants (F₄) with lower fertility, but not of plants with high fertility (F₁₀). GISH analysis indicated that these F₁₀ plants included the expected 18 chromosomes from R. sativus and B. alboglabra, respectively, but they lost approximately 27.6% R. sativus and 35.6% B. alboglabra AFLP (amplified fragment length polymorphism) bands. The crossability of the Raphanobrassica with R. sativus and 5 Brassica species (13 cultivars) were investigated. Seeds or F₁ seedlings were easy to be produced from crosses × Brassicoraphanus × R. sativus, and B. napus, B. juncea and B. carinata × Brassicoraphanus. Fewer seeds or seedlings were obtained from crosses × Brassicoraphanus × B. napus, B. juncea and B. carinata. However, few seeds were harvested in the reciprocals of × Brassicoraphanus with B. rapa and B. oleracea. The possible cause of fertility improvements and the potential of the present × Brassicoraphanus for breeding were discussed.     

Natural hybrids between cultivated and wild sunflowers (<Emphasis Type="Italic">Helianthus</Emphasis> spp.) in Argentina

Two introduced wild species Helianthus annuus L. and H. petiolaris Nutt. have become widespread in central Argentina and overlap the sunflower crop region. Intermediate off-type plants between the wild and cultivated species are often found, which is of concern because of the recent release of imidazolinone resistant varieties and the likely use of genetically modified sunflower cultivars. The progeny of 33 off-type plants obtained from 14 representative sites of the diffusion area were studied to confirm hybrid origin. Germination, survival, morphological traits and days to flowering confirmed hybridization between crop and both wild species, when compared to eight accessions of typical wild plants. Some progenies were presumably crop–wild H. annuus hybrids, some originated from the cross of cultivated plants and H. petiolaris, and two were the advanced generation of a cultivated hybrid. Hence, morphological traits are a good clue for the identification of spontaneous hybrid plants at field. The results indicate that crop–wild hybridization and introgression occur at various places in central Argentina. This fact may represent a way to herbicide resistance escape and future transgene escape if GM sunflower cultivars are released for commercial use.     

Genetic diversity and gene-pool subdivisions of diploid D-genome <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. (Poaceae) in Iran as revealed by AFLP

The diploid goatgrass Aegilops tauschii is considered the D-genome donor of bread wheat and has probably a centre of diversity in north of Iran. In order to measure the genetic diversity of and the relationships among different populations, varieties and subspecies belonging to Ae. tauschii in Iran, DNA was extracted from 48 accessions of Ae. tauschii collected across the geographic range of the species in the Country and the genetic diversity was assessed using AFLPs based on eight PstI/MseI +3 primer pairs resulted in 277 bands, 198 of which were polymorphic. High level polymorphism was detected, with an average of polymorphism rate of 0.715; relatively low genetic similarity (0.455) between accessions and significant difference between the lowest (0.179) and the highest genetic similarity (0.817). The Iranian Ae. tauschii populations showed high level of genetic diversity. The populations studied were divided into two groups: one group was mainly representing Northern populations collected from Southern Caspian Sea shore and the other group was mainly representing Northeast and Northwest populations. Based on the results of this study, it can be suggested that Ae. tauschii possesses two separate gene-pools in Iran: Northern and Northeastern–Northwestern. Considering the needs for introducing new characteristics and alleles for wheat improvement purposes, Ae. tauschii Iranian gene-pool is assumed to be of high importance for more investigation in the future.     

Effects of long-term mineral fertilization on microbial biomass,microbial activity,and the presence of <Emphasis Type="Italic">r</Emphasis>- and <Emphasis Type="Italic">K</Emphasis>-strategists in soil

Long-term effects of mineral fertilization on microbial biomass C (MBC), basal respiration (R _B), substrate-induced respiration (R _S), β-glucosidase activity, and the r–K-growth strategy of soil microflora were investigated using a field trial on grassland established in 1969. The experimental plots were fertilized at three rates of mineral N (0, 80, and 160 kg ha⁻¹ year⁻¹) with 32 kg P ha⁻¹ year⁻¹ and 100 kg K ha⁻¹ year⁻¹. No fertilizer was applied on the control plots (C). The application of a mineral fertilizer led to lower values of the MBC and R _B, probably as a result of fast mineralization of available substrate after an input of the mineral fertilizer. The application of mineral N decreased the content of C extracted by 0.5 M K₂SO₄ (C _ex). A positive correlation was found between pH and the proportion of active microflora (R _S/MBC). The specific growth rate (μ) of soil heterotrophs was higher in the fertilized than in unfertilized soils, suggesting the stimulation of r-strategists, probably as the result of the presence of available P and rhizodepositions. The cessation of fertilization with 320 kg N ha⁻¹ year⁻¹ (NF) in 1989 also stimulated r-strategists compared to C soil, probably as the result of the higher content of available P in the NF soil than in the C soil.     

Diversification of Multipurpose Plant, <Emphasis Type="Italic">Perilla Frutescens</Emphasis>

The traditional Asian crop, Perilla frutescens has multiple uses. There are specialized cultivars for seed oil and for medicinal use, as well as wild/weedy forms growing in various habitats. Based on selective characteristics of leaf odor, anthocyanin pigmentation, seed hardness and seed diameter, the diversity of this species was investigated to clarify the intraspecific differentiation. P. frutescens was divided into five groups by the combination of three qualitative characteristics: leaf odor, anthocyanin pigmentation and seed hardness. Most of the plants cultivated for oil belonged to one group, while medicinal plants belonged to three other groups. Wild/weedy forms were in the last group. The five groups could not be distinguished by seed diameter. Though the plants cultivated for oil tended to have larger seeds than the medicinal and wild/weedy plants, there was no boundary either between the two crops, or among various phenotypes of P. frutescens.     

Cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis> Crantz) genetic resources: a case of high iron and zinc

Cassava hybrids from interspecific crosses with Manihot caerulescens Pohl, M. pseudoglaziovii Pax and Hoffmann and M. dichotoma Ule showed a very high iron and zinc content in both roots and leaves, e.g. 98.15 mg kg⁻¹ in roots of the interspecific hybrid cassava-M. caerulescens versus 12.17 mg kg⁻¹ in a cassava cultivar. This promising results show the potential of wild Manihot species for micronutrient enhancement of cassava.     

Sources of wheat resistance to Sunn pest, <Emphasis Type="Italic">Eurygaster integriceps</Emphasis> Puton,in Syria

Sunn pest, Eurygaster integriceps Puton, is the most damaging insect pest of wheat in West and Central Asia and East Europe. Host plant resistance has been investigated as one component of a total integrated pest management program for the control of this pest. In Syria, field screening of artificially infested wheat accessions from the International Center for Agricultural Research in the Dry Areas (ICARDA) gene bank, selected using the Focused Identification of Germplasm Strategy (FIGS), identified one durum wheat and eight bread wheat accessions with good levels of resistance at the vegetative stage to overwintered Sunn pest adults. ICARDA is using these sources of resistance in wheat breeding programs to develop cultivars resistant to overwintered Sunn pest adults, which damage wheat at the vegetative stage (shoots and leaves). This study also demonstrated that the FIGS approach was effective in mining genetic resource collections for useful traits.     

Divergent evolution of wild and cultivated subspecies of <Emphasis Type="Italic">Triticum timopheevii</Emphasis> as revealed by the study of <Emphasis Type="Italic">PolA1</Emphasis> gene

Triticum timopheevii (genome symbol AAGG) comprises two subspecies, cultivated ssp. timopheevii, and wild ssp. armeniacum. These two subspecies are considered as allotetraploids of AA genome from Triticum diploid species and SS genome from Aegilops species. The difference in genome symbol (G vs. S) is due to wide genetic variations among four SS genome species, Ae. bicornis, Ae. longissima, Ae. searsii, and Ae. speltoides. In order to study the origin of T. timopheevii, we compared 19th intron (PI19) sequence of the PolA1 gene, encoding the largest subunit of RNA polymerase I. Two different sized DNA fragments containing PI19 sequences (PI19A and PI19G) were amplified both in ssp. timopheevii and ssp. armeniacum. Shorter PI19A (112 bp) sequences of both subspecies were identical to PI19 sequences of two AA species, T. monococcum and T. urartu. Interestingly, the longer PI19G (241–243 bp) sequences of ssp. armeniacum showed more similarity to PI19 sequences of Ae. speltoides whereas ssp. timopheevii showed more similarity to PI19 sequences of other three SS genome species. The results indicated that two subspecies of T. timopheevii, ssp. armeniacum or ssp. timopheevii, might have arisen independently by allotetraploidization of AA genome with Ae. speltoides or one of the remaining three Aegilops species, respectively.     

Identification of <Emphasis Type="Italic">SUB1A</Emphasis> alleles from wild rice <Emphasis Type="Italic">Oryza rufipogon</Emphasis> Griff.

Submergence stress is a major constraint to rice production in South and Southeast Asia. Most rice (Oryza sativa L.) cultivars die within a week of complete submergence, while a small number of accessions are submergence-tolerant for up to 2 weeks or more. These cultivars have the tolerant allele of the SUB1A gene, one of three ERF genes at this locus on rice chromosome 9. In all O. sativa varieties studied, the SUB1A gene is limited to a subset of indica accessions of O. sativa. Thus far, there has been no published report of the SUB1A gene in wild rice species. Here we report evidence of the SUB1A gene found in wild species of O. rufipogon Griff. accessions by the use of degenerate primers corresponding to the most highly conserved regions of the SUB1 locus. The results indicated that two SUB1A-like alleles, e.g. OrSub1A-1 and OrSub1A-2, were identified from two O. rufipogon accessions. Submergence treatment shows that both of the accessions with SUB1A-like genes were submergence-intolerant. This preliminary study provides insight into the origin and allelic variation of SUB1A, an agronomically important gene that is rapidly being introduced into widely-grown rice cultivars.     

Relationship of seedling and adult plant resistance and evaluation of wheat germplasm against tan spot (<Emphasis Type="Italic">Pyrenophora tritici</Emphasis>-<Emphasis Type="Italic">repentis</Emphasis>)

Genetic resistance is the most effective, economical and environment friendly method of managing tan spot of wheat caused by the ascomycete Pyrenophora tritici-repentis (Died.) Drechs. (anamorph Drechslera tritici-repentis, Died.). This study was carried out to determine the association between seedling and adult plant resistance in winter wheat cultivars, study the inheritance of tan spot resistance and evaluate wheat germplasm for resistance. A significant positive correlation was noted between seedling resistance evaluated in greenhouse and adult plant resistance estimated in field conditions. The absence of segregation into resistant plants in the F₁ disomic crosses of the resistant spring and winter wheat cultivars with the susceptible cultivars, and the segregation of the corresponding F₂ crosses into 1 resistant: 3 susceptible ratio indicated that tan spot resistance is controlled by a single recessive gene which inherits qualitatively. The winter wheat cultivars: Ibis, Heines VII, Albrecht, Solitar, Ohio, Toronto, Yindos, Zenith and Kronjuwel, and the spelt wheat cultivars: Ceralion, Hercule, and Schwabenkorn showed highly resistant response to both race 1 and race 5 isolates. We recommend these genotypes to be used for gene deployment in wheat breeding programs.     

Geographic distribution and domestication of wild emmer wheat (<Emphasis Type="Italic">Triticum dicoccoides</Emphasis>)

The transition from hunting and gathering to agriculture had revolutionary consequences for the development of human societies. Crops such as wheat, barley, lentil, pea and chickpea played a crucial role in the establishment of complex civilizations in south west Asia. Wild emmer wheat (Triticum dicoccoides) was one of the first cereals to be domesticated in the Fertile Crescent between c. 12,000 and c. 10,000 years ago. This step provided the key for subsequent bread wheat evolution. Wild emmer is found today in the western Fertile Crescent in Jordan, Syria and Israel, the central part of southeastern Turkey and mountain areas in eastern Iraq and western Iran. In this review, we summarize issues concerning geography and domestication of wild emmer wheat based on published molecular and archaeobotanical data and on our recent findings. We suggest that modern domestic tetraploid wheats derived from wild emmer lines from southeast Turkey. However, our understanding of emmer domestication is not complete. The “dispersed-specific” domestication model proposed for einkorn might well be appropriate also for emmer.     

Genetic and bioclimatic variation in <Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>

Solanum pimpinellifolium, due to its close relationship to S. lycopersicum, has been a genetic source for many commercially important tomato traits. It is a wild species found in the coastal areas of Peru and Ecuador. In this study, the genetic variation of S. pimpinellifolium was studied using the diversity found in 10 microsatellites in 248 plants spread throughout its entire distribution area, including Ecuador, which has been underrepresented in previous studies. Peruvian and Ecuadorian accessions are genetically quite differentiated. A possible cause of these differences could be the non-uniform nature of the coastal Ecuadorian and Peruvian climates, seeing as an important correlation between genetic differentiation and climate has been found. In addition, Ecuadorian and south Peruvian accessions have a lower genetic diversity and a higher homozygosity due to their higher autogamy, lower population size, and possible colonization bottlenecks. The Galápagos Islands population is an extreme case, with no diversity, likely caused by a recent colonization from the northern continental Ecuadorian region where genetically identical plants have been found. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Elena Zuriaga and José M. Blanca contributed equally to this work.     

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.