Genetic relationships within Vigna unguiculata (L.) Walp. based on isozyme analyses

Summary Isozyme analyses of genetic diversity in Vigna unguiculata were performed to determine genetic relationships and level of genetic diversity between wild and cultivated cowpea. Thirty-four cultivated accessions of V. unguiculata, 56 wild accessions of V. unguiculata, and six accessions representing five related wild Vigna species were analyzed. Ten enzyme systems were polymorphic within Vigna unguiculata: AAT, ACO, G6PDH, DIAP, LAP, MUE, ME MDH, PRX, and SOD. Fourteen of 24 putative loci (58%) were polymorphic within wild V. unguiculata, but only one locus (4%) was polymorphic within cultivated cowpea; when five related Vigna species were examined, 21 of the 24 bands of activity showed polymorphisms (88%) adding 33 alleles to the 48 identified within V. unguiculata. In one F₂ population of 68 plants (UCDVg 36 × UCDVg 21) a loose linkage was indicated between Diap-2 and G6pd-1 (² = 15.39; p = 0.004) with an estimated distance of 36.0 cM ± 5.02 (recombination (r) = 0.31). Also in another F₂ population of 38 plants (CB 88 × UCDVg 21) a loose linkage was indicated between Lap-1 and Prx (\gC² = 9.62; p = 0.047) with an estimated distance of 39.8 cM ± 7.0 (r = 0.33). Total genetic diversity (H_T) was 0.085 over all of the accessions including the one classified as V. nervosa. Within accession diversity (H_s) approached zero and between accession diversity (D_si) was responsible for all of the genetic diversity present. Therefore the coefficient of gene differentiation (G_ST = D_STII_T) approached 1. Absolute gene differentiation (D_m) was 0.087. Two of the nine segregations in this study were skewed. In general, results of this study concurred with the taxonomic classification within V. unguiculata and provided a strong indication that a severe genetic bottleneck occurred during the domestication process of cowpea.Abbreviations AAT aspartate amino-transferase - ACO aconitase - ALD aldolase - AUS Australia - BDI Burundi - BWA Botswana - CHN China - CMR Cameroon - DIAP diaphorase - DZA Algeria - ETH Ethiopia - G6PDH glucose-6-phosphate dehydrogenase - GDH glutamate dehydrogenase - GHA Ghana - GUY Guyana - IDH isocitrate dehydrogenase - IND India - KEN Kenya - LAO Laos - LAP leucine aminopeptidase - MDH malate dehydrogenase - ME malic enzyme - MEX Mexico - MOZ Mozambique - MUE methylumelliferyl-esterase - MWI Malawi - MYS Malaysia - NER Niger - NGA Nigeria - PRX peroxidase - RBSC ribulose-bisphosphate carboxylase - SEN Senegal - SLE Sierra Leone - SOD superoxide dismutase - TGO Togo - TZA Tanzania - USA United States of America - XDH xanthine dehydrogenase - ZAF South Africa - ZAR Zaire - ZIM Zimbabwe - ZMB Zambia     

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.     

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.     

Vigna vexillata (L.) A. Rich. Cultivated as a Root Crop in Bali and Timor

Vigna vexillata is considered as a pantropical distributed wild species closely related to the cowpea (Vigna uniculata) and adapted to infertile soils. The species is occasionally used for its storage roots as well as forage and erosion control plant. The objective of this study was to pursue personal communications that V. vexillata has be transformed into a cultivar in Indonesia. Seven Indonesian islands were visited (Java, Bali, Sumba, Flores, Timor, Kalimantan and Sulawesi). Wild V. vexillata was found in Java, Bali, Sumba, Flores and Timor, occasionally used as wild ‘forest food’, and cultivated V. vexillata was found in Bali and Timor, Seven cultivars were collected and two of these were made available for the National Botanic Garden of Belgium. The cultivars were primarily cultivated for their storage roots, propagated by seeds, required no scarified seeds for good germination and formed non-dehiscent pods. On-farm root yields of 18–30 t ha⁻¹ and seed yields of 0.7–1.2 t ha⁻¹ were estimated. A brief discussion about the common names of wild V. vexillata is given and it is proposed to use the name ‘tuber cowpea’ for V. vexillata accessions which are cultivated for their storage roots. The material may be of interest for regions, where the growing season is too short or the rainfall too low for sweet potato and cassava as well as to incorporate cultivar characteristics into wild V. vexillata accessions which are used for their storage roots.     

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.     

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.     

Karyomorphological and heterochromatin similarities among fourVigna species

Summary Karyomorphological similarities among four species of the genusVigna, V. unguiculata, V. luteola, V. oblongifolia andV. vexillata, were evaluated. Two different approaches were followed: cytomorphometrical determinations, carried out with an automated image analysis system to improve resolution, yielded some karyotypical indices; the distribution of heterochromatin bands was interpreted using a recently reported algorithm. Both methods led to similar results: the most karyologically similar species toV. unguiculata appears to beV. luteola, whileV. oblongifolia appears to be the least related one.     

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.     

Novel sources of resistance to the soybean cyst nematode (Heterodera glycines) found in wild relatives of azuki bean (Vigna angularis) and their characteristics of resistance

Soybean cyst nematode (Heterodera glycines; SCN) is becoming a serious problem in azuki bean (Vigna angularis var. angularis) production in Hokkaido, Japan. To find sources of resistance to SCN, we screened wild relatives of azuki bean and calculated “female indices” (FIs) of 342 accessions from 8 Vigna species that are cross-compatible with azuki bean. Twenty-three accessions belonging to V. hirtella, V. minima, V. nakashimae, V. riukiuensis, and V. tenuicaulis were resistant to the most prevalent SCN race, race 3, but most of them were only moderately resistant or susceptible to race 5. Four promising accessions (V. minima JP205886, JP205891, and JP210806, and V. nakashimae JP107879) showed a high level of resistance to all SCN races found in Japan (1, 3, and 5) and race 2 (an experimentally derived race from race 5). Since the SCN-resistant soybean cultivars released in Japan are not resistant to races 2 and 5, these wild Vigna accessions may have resistance mechanisms different from that of soybean. Continuous culture of race 3 on promising accessions showed that it might be difficult to overcome the resistance of these accessions. The resistance sources found in this study will be useful in controlling SCN through the breeding of SCN-resistant azuki bean cultivars. This study also showed the effectiveness of using wild genetic resources for identifying novel resistance sources.     

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.     

Genetic relationships and diversity among Brazilian cultivars and landraces of common beans (Phaseolus vulgaris L.) revealed by AFLP markers

The genetic variation and relationships among 31 accessions of Phaseolus vulgaris L., and two representatives of Vigna unguiculata L., were evaluated by AFLP analysis. A total of 263 DNA fragments across all materials were scored using nine primer combinations, averaging 32 per primer. More than 95% of the amplification products showed polymorphism, indicating high variation at the DNA level among these accessions. Pair-wise genetic similarity (Jaccard's coefficient) ranged from 0.553 to 0.840, with a mean of 0.765. Twenty-three accessions (70%) clustered into three groups. A majority of the commercial cultivars (91%) clustered within a single group, whereas the landraces were distributed along all the variation. An apparent correlation with phaseolin types was detected. Results of this study suggest that Brazilian landraces truly represent the overall genetic variability of Phaseolus vulgaris, confirming the multiple origins of these materials, and their potential as a source of variation for breeding programs.     

PCR Marker-based Analysis of Wild and Cultivated Yams (<Emphasis Type="Italic">Dioscorea</Emphasis> spp.) in Nigeria: Genetic Relationships and Implications for <Emphasis Type="Italic">ex situ</Emphasis> Conservation

Reliable characterization of the variation among wild and cultivated yams in Nigeria is essential for improved management and efficient utilization of yam genetic resources. RAPD and double stringency PCR (DS-PCR) analyses were used to investigate genetic relationships and the extent of redundancy among 30 accessions of two cultivated, and 35 accessions of four wild yam species collected from Nigeria. Twenty-five selected random decamer and two microsatellite primers were used individually and in combination to generate DNA profiles for each accession of the six Dioscorea species. The number of amplified fragments varied from 7 to 18 fragments per primer/primer combination. Different levels of intraspecific genetic diversity were found, with Dioscorea rotundata Poir. being the most variable. Based on identical profiles for the RAPD and DS-PCR primers, 12 duplication groups consisting of a total number of 37 accessions were observed in the present study. An UPGMA analysis grouped the majority of plants according to the species. Cultivated yams belonging to the D. cayenensis–rotundata species complex, which were classified into seven morphotypes/varietal groups, could be clearly separated into two major groups corresponding to D. rotundata Poir. and D. cayenensis Lam. D. cayenensis cultivars exhibited a low level of intraspecific variation and were genetically close to the wild species Dioscorea burkilliana J. Miège. D. rotundata cultivars classified into six varietal groups showed a high degree of DNA polymorphism and were separated into two major groups that appeared most closely related to Dioscorea praehensilis Benth. and Dioscorea liebrechtsiana de Wild. We propose, based on these results, that cultivars classified into D. cayenensis should be considered as a taxon separate from D. rotundata. The implications of intraspecific variability for the ex situ conservation of wild and cultivated yam germplasm in Nigeria are discussed.     

Assessment of genetic variation in a working collection of Vigna vexillata (L.) A. Rich. by isozyme and RAPD analyses

Genetic variation based on isozyme and RAPD analyses was investigated in 47 and 34 accessions respectively of Vigna vexillata from different geographical origins and belonging to three botanical varieties. A total of 9 enzyme systems were studied, accounting for 14 putative loci, 8 of which were polymorphic. The analysis of genetic diversity revealed a low level of within accession variation (H_S=0.013), while between accession diversity (D_ST) was 0.120. Coefficient of gene differentiation (G_ST) was 0.905, indicating that most variation was among accessions. Nei's genetic distances were calculated on the basis of allelic frequencies and a UPGMA dendrogram was constructed. Twenty arbitrary 10-mer oligonucleotides were used in RAPD analysis. Amplification profiles disclosed a higher level of polymorphism than isozymes. Based on amplification patterns, the similarity index of Jaccard was calculated and a dendrogram constructed on the basis of the similarity matrix. The final clustering based on RAPD data was similar to the one obtained using isozyme allelic frequencies. The classification in botanical varieties did not reflect the allelic constitution of the different samples. On the other hand, referring to geographical origin, most accessions from Africa and from Latin America were distributed respectively in two distinct clusters in the dendrogram. This grouping might also reflect the differences observed in the germination behaviour of V. vexillata from the two continents.     

AFLP analysis of diploid species in the genus Vigna subgenus Ceratotropis

A representative set of germplasm of genusVigna subgenus Ceratotropis specieswas used to analyze genetic diversity using amplified fragment lengthpolymorphism (AFLP) methodology. The germplasm was selected based onmorphological diversity and geographic origin and includes 18 out of the 21species in the subgenus Ceratotropis. Genetic diversitywithin and between taxa was measured using information generated by AFLP bandpolymorphism from which Jaccard's similarity coefficient and nucleotidediversity were estimated. The data were also used to generate phenograms tovisualize relationships among analyzed accessions. All species in sectionsCeratotropis and Aconitifoliae arewell separated. However, most of the species in sectionAngulares show a high level of similarity suggesting a lowlevel of genetic differentiation. In section Angulares onespecies, V. trinervia, is welldifferentiated from the other species and represents a separate evolutionarybranch between section Ceratotropis and other species insection Angulares. The relationships between the newlydescribed species V. aridicola,V. exilis, V.nepalensis and V.tenuicaulis and other species in the subgenusCeratotropis are described for the first time based on AFLPdata.     

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.     

A new taxonomic treatment for some wild relatives of mungbean (<Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilcz.) based on their molecular phylogenetic relationships and morphological variations

Mungbean (Vigna radiata (L.) Wilcz.) is an important leguminous crop cultivated mainly in Asia. Its wild relatives are considered useful genetic resources for mungbean breeding. However, the taxonomic history of mungbean and its wild relatives is complicated and some confusion is still present in recent publications. In this study, we examined the rDNA-ITS sequences and morphological characteristics of 83 gene bank accessions closely related to mungbean. As a result, we classified the 83 accessions into five species and one unclassified accession. The proper taxon name for each species was determined (Vigna grandiflora (Prain) Tateishi et Maxted, Vigna mungo (L.) Hepper, V. radiata, Vigna subramaniana (Babu ex Raizada) Raizada, and Vigna trinervia (Heyne ex Wight et Arn.) Tateishi et Maxted) based on a review of their taxonomic history and morphological comparisons between gene bank accessions and type specimens. A new taxonomic treatment is proposed and a morphological key has been prepared. In this treatment, Vigna radiata (L.) Wilcz. var. setulosa (Dalz.) Ohwi et Ohashi is treated as a synonym of Vigna radiata (L.) Wilcz. var. sublobata (Roxb.) Verdc., and Vigna hainiana Babu, Gopinathan et Sharma is a synonym of V. subramaniana. Accession ‘NI1135’ was revealed to be most closely related to, but is considerably differentiated from, mungbean (V. radiata) based on its rDNA-ITS sequences. It also has distinguishing morphological characters. Plants with similar morphologies and DNA profiles might be distributed in the Indian Himalaya. However, since only one accession is available at present, the taxonomic classification of ‘NI1135’ needs to be reconsidered in the future.     

Germplasm Sources for Improvement of Forage Quality in Lotus corniculatus L. and L. uliginosus Schkuhr (Fabaceae)

Extensive germplasm resources are present in collections of Lotus corniculatus and L. uliginosus, two forage species that are valuable in situations where other legumes are not persistent. Naturalised populations in many regions are the primary gene pool for genetic improvement of these species and interspecific hybrids are an important source of variation for agronomic and forage quality traits. Although previous research has provided evidence that selection for optimal levels of condensed tannins and herbage productivity are feasible breeding objectives, less is known about correlations of condensed tannin with other parameters of nutritive value. A germplasm collection of 38 accessions in both species from 9 geographic regions was used as a data base to determine these correlations. Forage of each accession was sampled in two successive years from a field site in south-eastern Australia and analysed for condensed tannin, in vitro digestible dry matter, nitrogen, acid detergent fibre and neutral detergent fibre. In both species condensed tannin was negatively correlated with in vitro digestible dry matter and nitrogen. This result helped to identify potentially valuable accessions with low condensed tannin (<4% of dry weight) and high in vitro digestible dry matter (>70%). One of these was an interspecific hybrid of L. uliginosus and L. corniculatus, confirming the view that interspecific hybridization would be a valuable avenue for improvement of agronomic and forage quality characters in Lotus species.     

Diversity of seed globulins in Lathyrus sativus L. and some related species

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of reduced seed globulins covered 141 accessions of Lathyrus sativus and the following related species: L. amphicarpos, L. blepharicarpus, L. cicera, L. gorgoni, L. marmoratus, L. pseudocicera and L. stenophyllus. In the total examined material 71 globulin polypeptide bands were distinguished. The number of polypeptide bands detected in particular species varied from 14 in L. amphicarpos to 29 in L. sativus; all the species under study showed intraspecific variation due to individual variation and/or differences among accessions. Electrophoretic data were subjected to statistical analysis using hierarchical UPGMA grouping. Frequency distribution of polypeptide bands in L. sativus showed some correlations with geographical origin and certain seed characteristics (seed coat colour, seed weight) of the studied accessions. As regards interspecific relationships, the studied species showed to be distantly related taxa except for the rather closely allied L. cicera and L. marmoratus. The obtained results are compared with the electrophoretic seed albumin data reported earlier for the same taxa.     

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.     

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.