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.     

Analysis of genetic diversity in a sweet potato (Ipomoea batatas L.) germplasm collection from Tanzania as revealed by AFLP

Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.     

Diversity and genetic resources of wild <Emphasis Type="Italic">Vigna</Emphasis> species in India

Diversity in morphological characters of 206 accessions of 14 wild Vigna species from India was assessed. Of these, 12 species belonged to Asian Vigna in the subgenus Ceratotropis and two were V. vexillata and V. pilosa belonging to subgenus Plectotropis and Dolichovigna, respectively. Data on 71 morphological traits, both qualitative and quantitative, were recorded. Data on 45 qualitative and quantitative traits exhibiting higher variation were subjected to multivariate analysis for establishing species relationships and assessing the pattern of intraspecific variation. Of the three easily distinguishable groups in the subgenus Ceratotropis, all the species in mungo-radiata group, except V. khandalensis, viz. V. radiata var. sublobata, V. radiata var. setulosa, V. mungo var. silvestris and V. hainiana showed greater homology in vegetative morphology and growth habit. The species, however, differed in other plant, flower, pod and seed characteristics. Within species variation was higher in V. mungo var. silvestris populations and three distinct clusters could be identified in multivariate analysis. V. umbellata showed more similarity to V. dalzelliana than V. bourneae and V. minima in the angularis-umbellata (azuki bean) group. Within species variations was higher in V. umbellata than other species in the group. In the aconitifolia-trilobata (mothbean), V. trilobata populations, were more diverse than V. aconitifolia. The cultigens of the conspecific wild species were more robust in growth, with large vegetative parts and often of erect growth with three- to five-fold increase in seed size and seed weight, except V. aconitifolia, which has still retained the wild type morphology to a greater extent. More intensive collection, characterisation and conservation of species diversity and intraspecific variations, particularly of the close wild relatives of Asian Vigna with valuable characters such as resistance to biotic/abiotic stresses, more number of pod bearing clusters per plant etc. assumes great priority in crop improvement programmes.     

AFLP Analysis of the Phenetic Organization and Genetic Diversity in the Sugarcane Complex, Saccharum and Erianthus

Amplified fragment length polymorphism (AFLP) markers were evaluated for determining the phylogenetic relationships, and the diversity in the Saccharum complex using 30 clones belonging to S. officinarum, S. robustum, S. spontaneum, S. barberi, S. sinense and the related genus Erianthus. The phenetic tree of the species clones based on AFLP data was consistent with the known taxonomical relationships. AFLP gave higher resolution of closely related species into discrete groups than that by RAPD and RFLP markers, reported earlier. The levels of diversity within the various Saccharum species were also found to be higher than those obtained previously with the same set of clones using RAPD markers. The intraspecies similarity in S. barberi and S. sinense was much higher than interspecies similarity suggesting a clear separation of the two, which are considered ‘horticultural species’. The genetic similarity matrix derived from a single primer combination highly correlated (r = 0.980) with that obtained from all the 12 primer combination used in the study, thus highlighting the efficiency of a single primer combination in delineating species relationships. All the primer combinations could identify markers that are specific to each of the species and the genus Erianthus. Among the species, specific markers were highest in S. spontaneum followed by S. robustum, S. barberi, S. officinarum and S. sinense. Erianthus had a distinct profile with 30% of the total amplified fragments being specific to it. This offers great scope for identifying intergeneric hybrids, which has been very difficult using morphological traits and RAPD markers. High degree of correspondence between the results from the cluster analysis based on Jaccard's similarity index, Neighbour Joining tree based on Sokal and Michener distance matrix and AFTD (Analyses Factorielle on Table of Distances) analysis clearly demonstrated that AFLP markers would be an appropriate tool in providing better information about the relationships among the species, estimation of diversity, and in revealing species and genus specific markers that could be directly applied in sugarcane breeding programmes.     

Genetic Variation and Relationships of Pedigree-Known Oat, Wheat, and Barley Cultivars Releaved by Bulking and Single-Plant Sampling

Applications of bulking procedures have played an increasingly important role in molecular characterization of plant germplasm, but little attention has been made to address the effectiveness of detecting genetic variation and inferring genetic relationships via bulking. An analysis was performed here to compare the genetic variation detected and genetic relationships inferred via bulking and single-plant sampling of five oat (Avena sativa L.), wheat (Triticum aestivum L.), and barley (Hordeum vulgare L.) cultivars with known pedigrees using amplified fragment length polymorphism (AFLP) markers. Three AFLP primer pairs were applied to screen one bulk and eight single-plant samples of each cultivar and up to 140 AFLP bands were scored for each sample. Analyses of these AFLP data showed bulking revealed AFLP variation up to 21.4% less than corresponding single-plant sampling for these crop species and also introduced up to 2.2% upward and 5.1% downward biases in detecting AFLP variations for each cultivar. The genetic relationships inferred by bulking using the Dice's coefficient, the simple matching coefficient, and the Jaccard's coefficient were largely the same, but differed from those in single-plant samples employing average Dice's coefficient, average simple matching coefficient, and AMOVA-based distance method. All of the inferred genetic relationships were not congruent to the known pedigrees. Clearly, substantial biases could exist in detection of AFLP variation and in inference of genetic relationships from bulk samples, even for closely related germplasm, and more efforts to assess the effectiveness of bulking in inferring genetic variation and relationships are needed for more informative molecular characterization of plant germplasm.     

Genetic variation within and among species of five sections of the genus Arachis L. (Leguminosae) using RAPDs

Some Arachis species are widely used as commercial plants, e.g. the groundnut A. hypogaea, an important source of good quality protein and oil, and A. pintoi and A. glabrata, that are utilized as forage species. Germplasm of most Arachis species is available in germplasm banks. However, little it is known about the genetic attributes of this germplasm, and mainly about its genetic variability, which is very important for its maintenance. In the present study RAPDs were used to assay the genetic variation within and among 48 accessions of five sections of the genus Arachis and to establish the genetic relationships among these accessions. Ten of 34 primers tested were selected for DNA amplification reactions since they yielded the largest numbers of polymorphic loci. A dendrogram was constructed based on data from the 10 primers selected. Eighty RAPD polymorphic bands were analyzed among the accessions studied. The relationships among species based on RAPDs were similar to those previously reported based on morphological, cytological and crossability data; demonstrating that RAPDs can be used to determine the genetic relationships among species of the different sections of the genus Arachis. In general, wide variation was found among accessions and low variation was found within the accessions that had two or more plants analyzed. However, higher polymorphism was found in the section Trierectoides and in one accession of A. major, indicating that generalizations should be avoided and each species should be analyzed in order to establish collection and maintenance strategies.     

Highly polymorphic AFLP markers as a complementary tool to ITS sequences in assessing genetic diversity and phylogenetic relationships of sweetpotato (Ipomoea batatas (L.) Lam.) and its wild relatives

Comparative analyses of genetic diversity and phylogenetic relationshipsof sweetpotato (Ipomoea batatas (L.) Lam.) and its wildrelatives in Ipomoea series Batataswere conducted using amplified fragment length polymorphism (AFLP) and sequencedata from the internal transcribed spacer (ITS) region of the ribosomal DNA. LowITS divergence among thirteen species of ser. Batatasresulted in poorly resolved relationships. More variable AFLP characters werefound to be more efficient in characterizing genetic diversity and phylogeneticrelationships at both intra- and interspecific levels within ser.Batatas. Highly informative AFLP fingerprints of 36accessions representing 10 species of ser. Batatas weregenerated using only six primer combinations. Of the species examined,I. trifida was found to be the mostclosely related to I. batatas, whileI. ramosissima andI. umbraticola were the most distantlyrelated to I. batatas. The highlypolymorphic AFLP markers are a valuable tool in assessing genetic diversity andphylogenetic relationships of sweetpotato and its wild relatives.     

An AFLP diversity study of the genus Passiflora focusing on subgenus Tacsonia

DNA variation was studied at the intra- andinterspecific levels within the genus Passiflora, with particular emphasis on the subgenusTacsonia and its relationship with the subgenusPassiflora. AFLP fingerprints were generated for35 accessions from 21 species, 14 of which belong to subgenusTacsonia, and four to subgenusPassiflora. The morphologically intermediateP. manicata (subgenusManicata) was also included, as wereP. trinervia (subgenusPsylanthus) and P.alnifolia (subgenus Decaloba). The two latter were used as outgroup species. Twoprimer combinations generated 34–80 scorable fragments pergenotype for a total of 260 fragments. Polymorphism analyses spannedthe levels from individual genotype to section. SubgenusTacsonia presented wide genetic variability atboth intra- and interspecific levels, particularly within wildspecies. The classification based on these markers generally agreeswell with current taxonomic definitions of common species, but lesswith definitions of sections. A geographical component was evident inthe intraspecific variation of P.mixta, P.tripartita var. mollissimaand P. tarminiana, withwider variation in Ecuador and Peru than in Colombia and Venezuela.These same three species, plus P.cumbalensis, formed adjacent clusters, showingcloser affinity with each other than with the other species ofsubgenus Tacsonia. As expected, P. manicata clustered at anintermediate position between subgenera Tacsoniaand Passiflora.     

Genetic Variation of Ethiopian Mustard (Brassica carinata A. Braun) Germplasm in Western Canada

Information on genetic diversity and genetic relationships among genotypes of Brassica carinata is currently limited. The objectives of this study were to evaluate patterns and levels of genetic diversity in B. carinata based on amplified fragment length polymorphisms (AFLP) as compared with Brassica juncea and Brassica nigra, and to evaluate agronomic and seed quality data for plants grown in the field in western Canada. A total of 296 AFLP bands were generated from four primer pair combinations and scored for presence/absence in 66, 20 and 7 accessions of B. carinata, B. juncea and B. nigra, respectively. B. carinata was less genetically diverse than the other two species. Differences in diversity were evident in the proportion of polymorphic loci within each species: 23, 35 and 50% for B. carinata, B. nigra and B. juncea, respectively. Pair-wise similarity measures based on the Jaccard coefficient were highest among accessions of B. carinata and showed the narrowest range: 0.911 (0.810–0.981) compared to B. nigra: 0.569 (0.438–0.660) and B. juncea: 0.715 (0.345–0.951). AFLP-based genetic distance information can be used by plant breeders to select diverse genotypes. AFLPs are also useful for fingerprinting cultivars and two primer pair combinations were sufficient to uniquely identify all the accessions of B. carinata. More variation among accessions was identified in the agronomic trial than had previously been described in studies of B. carinata in western Canada, but the data were too limited to draw conclusions regarding specific accessions. Overall, the findings were in agreement with other published work describing the favourable agronomic potential of this species.     

AFLP Analysis of Genetic Diversity in Indian Soybean [Glycine max (L.) Merr.] Varieties

AFLP technique was used to assess genetic diversity in 72 soybean varieties under cultivation in India. Selected 12 AFLP primer pairs produced 1319 products of which 1257 were polymorphic (95%). Wide variations were observed for the number of amplification products, percent polymorphism and average polymorphism information content (PIC). The Jaccard's similarity indices (J) based on the AFLP profiles of the 72 soybean varieties were subjected to UPGMA cluster analysis. The dendrogram generated revealed four major clusters, which were strongly supported by the high bootstrap values obtained from analyses of 1000 bootstrap samples. In addition, the Mantel's test for cophenetic correlation with r = 0.955 indicated very good fit of the varieties to a group in the cluster analysis. Some correspondence between the clustering pattern and the pedigree, place of release or target area of the variety was observed. Overall moderately high genetic diversity was observed which appears to be due to the higher genetic diversity prevalent in 12 of the varieties included in three diverse clusters and was indicative of the need to include more diverse germplasm in the soybean improvement programs.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

A-genome cotton as a source of genetic variability for Upland cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis>)

Since Upland cotton (Gossypium hirsutum) is known to have relatively low levels of genetic diversity, a better understanding of variation and relationships among possible sources of novel genes would be valuable. Therefore, analysis of genetic variation of the genus Gossypium, especially the diploids, which are the putative donors of the A and D genomes for the commercially important allotetraploid cottons (AADD), G. hirsutum and G. barbadense, could provide important information about the feasibility of using these genetic resources for cotton improvement. The primary objective of this study was to analyze the genetic diversity in A-genome diploid cotton species, G. herbaceum (A1) and G.␣arboreum (A2) by using microsatellite markers. Forty-one A-genome germplasm accessions were evaluated with 32 microsatellite loci. Genetic similarities between A1 and A2 ranged from 0.62 to 0.86 with a mean of 0.70. Within each A-genome species similarities ranged from 0.80 to 0.97 with a mean of 0.89 for A1 and from 0.82 to 0.98 with a mean of 0.89 for A2. A UPGMA tree and principal coordinate analysis based on genetic similarity matrices showed distinct clusters consistent with the genomic groups.     

Intra-specific DNA polymorphism in pineapple (<Emphasis Type="Italic">Ananas comosus</Emphasis> (L.) Merr.) assessed by AFLP markers

Pineapple (Ananas comosus (L.) Merr.) cultivars, often derived from somatic mutations, are propagated vegetatively. It has been suggested by isozyme data that there is little genetic variation among Smooth Cayenne cultivars. A thorough investigation of the genetic variation within the cultivated speciesAnanas comosus, particularly among commercial cultivars, will provide critical information needed for crop improvement and cultivar protection. One-hundred and forty-eight accessions ofA. comosus and 14 accessions of related species were evaluated with AFLP markers. The average genetic similarity ofA. comosus was 0.735 ranging from 0.549 to 0.972, suggesting a high degree of genetic variation within this species. With AFLP markers, discrete DNA fingerprints were detected for each commercial cultivar, breeding line, and intra-specific hybrid. Self-incompatibility, high levels of somatic mutation, and intraspecific hybridization may account for this high degree of variation. However, major cultivar groups of pineapple, such as Cayenne, Spanish, and Queen, could not be distinctively separated. These cultivar groups are based on morphological similarity, and the similar appearance can be caused by a few mutations that occurred on different genetic background. Our results suggest that there is abundant genetic variation within existing pineapple germplasm for selection, and discrete DNA fingerprinting patterns for commercial cultivars can be detected for cultivar protection. The genetic diversity and relationships of fourAnanas species are also discussed.     

Genetic relationships among perennial and annual Cicer species growing in Turkey as revealed by allozymes

Allozyme polymorphisms were used to assess genetic variation and relationships among ten Cicer species (annuals and perennials) growing in Turkey. Using seven enzyme systems, 12 putative scorable loci were detected and surveyed for polymorphism in an accession collection including wild and cultivated forms. Variation was generally low within accessions and species, but common between species. Cluster analysis based on the pairwise genetic distance coefficients among accessions and species using UPGMA revealed two species clusters; one includes three perennials (C. montbretii, C. isauricum and C. anatolicum) and the other contains six annuals (C. pinnatifidum, C. bijugum, C. judaicum, C. echinospermum, C. reticulatum and C. arietinum) and one perennial species (C.incisum). Grouping obtained in allozyme analysis appears to be consistent with the classification these species into three sections. However, contrary to relationships obtained in previous studies, three perennial species from section Polycicer were relatively distant to the group containing annuals. One perennial species, C. incisum from section Chamaecicer, clustered with annuals showing a close similarity. The grouping of six annual species was consistent with the previous reports of relationships. The relationships deduced between perennials and annuals appear to shed light on the evolution of annual habit from perennial habit.     

RAPD and AFLP assessment of genetic variation in a landrace of pepper (Capsicum annuum L.), grown in North-West Italy

In several regions of Italy as well as other parts of southern Europe, the heterogeneity of the land, the climate and the soil favour the survival in cultivation of a large number of landraces specifically adapted to local conditions. Knowledge on the level and distribution of their genetic variation can help to develop appropriate strategies, in order to suistainably manage in situ these germplasm resources at risk of genetic erosion. C. annuum is an herbaceous diploid species and is considered to be self-pollinating, although different rates of out-crossing have been recorded. We used random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers to assess genetic diversity within and between five populations of a landrace of Capsicum annuum L., grown in a limited area in north-west Italy and locally known as Cuneo pepper. Partitioning the genetic variation with Shannon's diversity index revealed that 41.6% occurred between and 58.4% within populations. Analogous results were obtained when the analysis was based only on RAPD or AFLP markers. However, AFLP was more reliable, since a lower range of variation was observed among primer combinations in detecting the two components of genetic variation. Notwithstanding the rather high level of within genetic variation detected, the five populations were clearly differentiated and differed in the frequency of alleles exclusive and/or present at very low frequencies. Our results show the need for accurate estimation of allele frequencies, in order to identify populations to which priority should be given for dynamic conservation of landraces.     

Genetic similarities among Spanisch populations of Agropyron, Elymus and Thinopyrum, using PCR-based markers

Arbitrary oligonucleotides were used as primers to amplifygenomic DNA of 48 wild Spanish populations of Agropyroncristatum, Elymus hispanicus,E. caninus,E. repens,Thinopyrum curvifolium, Th.junceum and Th.intermedium. Genetic diversity was analysedusing nineteen primers. The number of amplified products ranged from8 to 18 per primer and a total of 240 markers were scored. Differentlevels of intraspecific genetic diversity were found, the allogamousspecies E. repens andTh. intermedium being themost variable. Jaccard's similarity coefficients for internalmeasure within and between populations were used to produce a clusterdiagram. The results demonstrate differences in the degree ofsimilarity between taxonomic units. Interpopulational variability andinterspecific genomic relationships of these species arediscussed.     

Phylogenetic relationships within <Emphasis Type="Italic">Citrus</Emphasis> and its related genera as inferred from AFLP markers

We investigated the phylogenetic relationships among Citrus and its relatives, including 29 genotypes belonging to Citrus, Poncirus, Fortunella, Microcitrus, Eremocitrus, Atalantia and Severinia using AFLP analysis. Cluster analysis was conducted using neighbour joining (NJ) method. The results demonstrated that Poncirus, Microcitrus and Eremocitrus are distant from Citrus. A strong affiliation exists between C. halimii and Fortunella and the results did not support C. halimii as the fourth basic species. Further evidence was provided that P. polyandra should not be the hybrid between trifoliate orange and Citrus genotypes and it deserves species status. Neither the separation of subgenus Papeda and Citrus nor the separation of subgenus Archicitrus and Metacitrus were clearly resolved in the dendrogram. C. ichangesis is a distinct species very different from other Citrus genotypes, and the results showed it is improper to classify it into subgenus Metacitrus. C. reticulata, C. maxima and C. medica were separated into three distinct clusters. Therefore if only cultivated Citrus species are considered, the three basic species concept is acceptable. However, it is not applicable for whole genus Citrus.     

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.     

Characterisation of germplasm accessions of Napier grass (pennisetum purpureum and P. purpureum×P. glaucum Hybrids) and comparison with farm clones using RAPD

Fifty six germplasm accessions of the important East African fodder crop Napier grass, Pennisetum purpureum, and its hybrids with P. glaucum, were characterised using 67 random amplified polymorphic DNA (RAPD) fragments. No or very low intra-accession variation was found for 49 of the accessions examined, confirming field observations that this species is predominantly clonally propagated. Comparison of intra and inter-accession variation identified several groups of identical/similar accessions that could be targeted if the collection is to be rationalised, and also highlighted two misplantings of germplasm material during transfer to a field trial site. A neighbour joining dendrogram of Jaccard's similarity estimates, clearly separated 50 accessions of P. purpureum from three P. glaucum individuals, and placed six hybrid accessions in an intermediate position. These groupings were well supported by a nested AMOVA (P<0.001; 29.5% of total variance due to taxonomic delineation). The main group of P. purpureum individuals could be further differentiated into five sub-groups (designated East Africa, Southern Africa, USA1, USA2 and Miscellaneous, to reflect the majority membership of sub-groups) and examination of the within P. purpureum component of the nested AMOVA, found them to be significantly different (P<0.001; 18.8% of variance). Genetic diversity across all accessions was found to be fairly high (Shannon's diversity index 0.306) and thus the collection probably represents a wide genetic base for this species. In addition to germplasm accessions, 25 Kenyan farm clones were also analysed. A principal coordinate analysis found that all but one of the clones clustered with the main P. purpureum group of accessions, indicating that the majority are probably not interspecific hybrids. The origin and pedigree of clones is discussed based on genetic similarity amongst clones and to germplasm accessions.     

Genetic variation analysis in the genus Lathyrus using RAPD markers

RAPD analysis was applied to five species belonging to the genusLathyrus (Fabaceae): L.sativus, L.cicera, L.ochrus, L.sylvestris and L.latifolius. All the species under study belong to thesection Lathyrus except L.ochrus which is in section Clymenum.Nine populations representing these species were used and ten random10-mer primers were sampled. A total of 129 amplification products,ranging in size from 0.3 to 3 Kb, were generated. Partitioning ofvariation was studied using the AMOVA technique. The genetic variation proved tobe nearly equally distributed among species and among populations withinspecies. A similar approach was carried out to distinguish populations, whichproved to be efficient. The between population dissimilarities were calculatedand a dendrogram of genetic relationships was drawn. The RAPDs obtained weresufficient to distinguish between the two accessions of a species and toseparate these accessions by clustering them according to species. A highgenetic similarity between the populations of L.sylvestris and L.latifolius was established. A similar result is also shownfor the populations of L. sativus andL. cicera.