Molecular taxonomic relationships in the genus Vigna based on RFLP analysis

Summary The taxonomy of the genus Vigna has been primarily based on morphological attributes. We have used 27 genomic clones from soybean, common bean, mungbean and cowpea to examine restriction fragment length polymorphism (RFLP) among 44 accessions of different species belonging to four subgenera of the genus Vigna. One accession each of common bean (Phaseolus vulgaris) and soybean (Glycine max) was included in the study. Total DNA from the various genotypes was digested with one restriction enzyme (Eco RV). Results of a numerical taxonomic analysis showed a high level of genetic variation within the genus with a remarkably higher amount of variation associated with Vigna sp. from Africa relative to those from Asia. The distinctness of the Asiatic grams in subgenus Ceratotropis, cowpea in section Catiang, bambara groundnut (V. subterranean) and members of the subgenus Plectotropis was elucidated by this study. Members of the subgenus Plectotropis were closer in genome homology to those of subgenus Vigna section Catiang than to those of subgenus Ceratotropis. The relative positions of some genotypes to one another on the dendrogram and minimum spanning tree were discussed in regard to hybridisations aimed generating well-saturated genomic maps and interspecies transfer of desirable genes.     

Novel Genetic Resources in the Genus Vigna Unveiled from Gene Bank Accessions

The genus Vigna (Fabaceae) consists of five subgenera, and includes more than 100 wild species. In Vigna, 10 crops have been domesticated from three subgenera, Vigna, Plectrotropis, and Ceratotropis. The habitats of wild Vigna species are so diverse that their genomes could harbor various genes responsible for environmental stress adaptation, which could lead to innovations in agriculture. Since some of the gene bank Vigna accessions were unidentified and they seemed to be novel genetic resources, these accessions were identified based on morphological traits. The phylogenetic positions were estimated based on the DNA sequences of nuclear rDNA-ITS and chloroplast atpB-rbcL spacer regions. Based on the results, the potential usefulness of the recently described species V. indica and V. sahyadriana, and some wild Vigna species, i.e., V. aconitifolia, V. dalzelliana, V. khandalensis, V. marina var. oblonga, and V. vexillata, was discussed.     

Genetic variability and relationships between and within grape cultivated varieties and wild species based on SRAP markers

Sequence-related amplified polymorphism (SRAP) markers were used to assess genetic relationships among 76 grape genotypes including Chinese indigenous and newly bred varieties, representatives of foreign grape varieties, and wild Vitis species. Nineteen informative primers were selected from 100 SRAP primer pairs due to their ability to produce clearly and repeatedly polymorphic and unambiguous bands among the varieties. A total of 228 bands were produced; 78.63% of them were polymorphic; the average polymorphism information content (PIC) is 0.76. Genetic relationships were obtained using Nei and Li similarity coefficients. Cluster analysis of SRAP markers through the unweighted pair-group method of arithmetic averages (UPGMA) analysis and principal coordinate analysis (PCoA) were largely consistent. The definition of clusters in the dendrogram and PCoA plot is the same and some degree of grouping by types of grape, ecogeographical origin, and taxonomic status of the varieties was revealed. Three main groups were found after cluster analysis, i.e., table grape of Vitis vinifera; table grape of Euro-America hybrid and wine grape of V. vinifera; wild Vitis species. Groupings indicated a divergence between the table and wine-type varieties of V. vinifera. The results showed that the wild Vitis species that originated from America and China could be clearly differentiated and Vitis hancockii is the most distant from the others of Asian Vitis species. The results also indicated that SRAP markers are informative and could distinguish bud sports of grape. The present analysis revealed that Chinese cultivated and wild grape germplasm are highly variable and have abundant genetic diversity.     

Chromosome numbers in the genus Mimosa L.: cytotaxonomic and evolutionary implications

Chromosome numbers were determined for 125 accessions of 92 taxa of Mimosa from all five of Barneby??s (Mem New York Bot Gard 65:1?C835, 1991) taxonomic sections. For 69 species, 1 subspecies and 8 varieties, chromosome numbers are presented for the first time, for 6 species and 1 variety previously published data have been confirmed and for 3 species and 2 varieties different numbers were found. Results show that 74% of the accessions were diploid (2n?=?2x?=?26) and 26% polyploid, these mostly tetraploid (2n?=?4x?=?52) but with two triploid (2n?=?3x?=?39). These results double the number of Mimosa species for which the chromosome count is known from less than 10% previously reported to more than 20%, representing an important advance in the cytotaxonomy of this legume genus. These results together with literature data show that ca. 78% of Mimosa species are diploid. Polyploids are present in most of the taxonomic sections and in different lineages across the genus. No particular chromosome number is restricted to a given section or lineage. A possible relation between geography, species distribution, polyploidy and invasiveness was detected, however, further studies based on more accessions, especially from higher latitudes, are required before firm conclusions can be drawn.     

Genome-wide single nucleotide polymorphism discovery and validation in adzuki bean

Adzuki bean, also known as red bean (Vigna angularis), with 2n = 22 chromosomes, is an important legume crop in East Asian countries, including China, Japan, and Korea. For single nucleotide polymorphism (SNP) discovery, we used Vigna accessions, V. angularis IT213134 and its wild relative V. nakashimae IT178530, because of the lack of DNA sequence polymorphism in the cultivated species. Short read sequences of IT213134 and IT178530 of approximately 37 billion and 35 billion bp were produced using the Illumina HiSeq 2000 system to a sequencing depth of 61.5× and 57.7×, respectively. After de novo assembly was carried out with trimmed HiSeq reads from IT213134, 98,441 contigs of various sizes were produced with N50 of 13,755 bp. Using Burrows–Wheeler Aligner software, trimmed short reads of V. nakashimae IT178530 were successfully mapped to IT213134 contigs. All sequence variations at the whole-genome level were examined between the two Vigna species. Of the 1,565,699 SNPs, 59.4 % were transitions and 40.6 % were transversions. A total of 213,758 SNPs, consisting of 122,327 non-synonymous and 91,431 synonymous SNPs, were identified in coding sequences. For SNP validation, 96 SNPs in the genic region were chosen from among IT213134 contigs longer than 10 kb. Of these 96 SNPs, 88 were confirmed by Sanger sequencing of 10 adzuki bean genotypes from various geographic origins as well as IT213134 and its wild relative IT178530. These genome-wide SNP markers will enrich the existing Vigna resources and, specifically, could be of value for constructing a genetic map and evaluating the genetic diversity of adzuki bean.     

Abundant Microsatellite Diversity and Oil Content in Wild Arachis Species

The peanut (Arachis hypogaea) is an important oil crop. Breeding for high oil content is becoming increasingly important. Wild Arachis species have been reported to harbor genes for many valuable traits that may enable the improvement of cultivated Arachis hypogaea, such as resistance to pests and disease. However, only limited information is available on variation in oil content. In the present study, a collection of 72 wild Arachis accessions representing 19 species and 3 cultivated peanut accessions were genotyped using 136 genome-wide SSR markers and phenotyped for oil content over three growing seasons. The wild Arachis accessions showed abundant diversity across the 19 species. A. duranensis exhibited the highest diversity, with a Shannon-Weaver diversity index of 0.35. A total of 129 unique alleles were detected in the species studied. A. rigonii exhibited the largest number of unique alleles (75), indicating that this species is highly differentiated. AMOVA and genetic distance analyses confirmed the genetic differentiation between the wild Arachis species. The majority of SSR alleles were detected exclusively in the wild species and not in A. hypogaea, indicating that directional selection or the hitchhiking effect has played an important role in the domestication of the cultivated peanut. The 75 accessions were grouped into three clusters based on population structure and phylogenic analysis, consistent with their taxonomic sections, species and genome types. A. villosa and A. batizocoi were grouped with A. hypogaea, suggesting the close relationship between these two diploid wild species and the cultivated peanut. Considerable phenotypic variation in oil content was observed among different sections and species. Nine alleles were identified as associated with oil content based on association analysis, of these, three alleles were associated with higher oil content but were absent in the cultivated peanut. The results demonstrated that there is great potential to increase the oil content in A. hypogaea by using the wild Arachis germplasm.     

Detection of Genome Donor Species of Neglected Tetraploid Crop Vigna reflexo-pilosa (Créole Bean), and Genetic Structure of Diploid Species Based on Newly Developed EST-SSR Markers from Azuki Bean (Vigna angularis)

Vigna reflexo-pilosa, which includes a neglected crop, is the only one tetraploid species in genus Vigna. The ancestral species that make up this allotetraploid species have not conclusively been identified, although previous studies suggested that a donor genome of V. reflexo-pilosa is V. trinervia. In this study, 1,429 azuki bean EST-SSR markers were developed of which 38 EST-SSR primer pairs that amplified one product in diploid species and two discrete products in tetraploid species were selected to analyze 268 accessions from eight taxa of seven Asian Vigna species including V. reflexo-pilosa var. glabra, V. reflexo-pilosa var. reflexo-pilosa, V. exilis, V. hirtella, V. minima, V. radiata var. sublobata, V. tenuicaulis and V. trinervia to identify genome donor of V. reflexo-pilosa. Since both diploid and tetraploid species were analyzed and each SSR primer pair detected two loci in the tetraploid species, we separated genomes of the tetraploid species into two different diploid types, viz. A and B. In total, 445 alleles were detected by 38 EST-SSR markers. The highest gene diversity was observed in V. hirtella. By assigning the discrete PCR products of V. reflexo-pilosa into two distinguished genomes, we were able to identify the two genome donor parents of créole bean. Phylogenetic and principal coordinate analyses suggested that V. hirtella is a species complex and may be composed of at least three distinct taxa. Both analyses also clearly demonstrated that V. trinervia and one taxon of V. hirtella are the genome donors of V. reflexo-pilosa. Gene diversity indicates that the evolution rate of EST-SSRs on genome B of créole bean might be faster than that on genome A. Species relationship among the Vigna species in relation to genetic data, morphology and geographical distribution are presented.     

Serological evidence confirming the assignment of phaseolus aureus and P. mungo to the genus vigna

Endopeptidase activity was detected in extracts of cotyledons of 11 species of Vigna and Phaseolus Antibodies against the purified protease isolated from the cotyledons of 5-day-old P.aureus seedlings inhibited the activity of that enzyme in crude extracts of cotyledons. A similar inhibition was obtained with P. mungo, P. adenanthus and 4 species of Vigna, while there was no inhibition of endopeptidase activity in extracts of cotyledons of 4 species of Phaseolus. Immunodiffusion tests proved that the protease of Vigna is distinct from that of Phaseolus. The evidence supports the reassignment of P. aureus and P. mungo to the genus Vigna and indicates that the names Vigna radiata and Vigna mungo are more appropriate than P. aureus and P. mungo for green gram and black gram respectively.     

Molecular characterization of intergenic spacer region of 5S ribosomal RNA genes in subgenus Vigna: extensive hybridization among V. unguiculata subspecies

We report the molecular analysis of the 5S ribosomal RNA intergenic spacer (IGS) region from 57 Vigna species of subgenus Vigna. Sequence analysis revealed that the 5S IGS was highly variable in length (189?C237?bp) and sequence (58% polymorphic sites). Most of the Vigna species analysed harboured a single type of 5S rRNA repeat unit, except V. unguiculata and V. reticulata that showed multiple ??intragenomic?? 5S types. The intragenomic 5S types among the six V. unguiculata subspecies were characterized by PCR-RFLP, genomic RFLP and sequencing. The 5S IGS was phylogenetically informative (comparable to ITS-1 and ITS-2 spacers) in inferring species relationships among the Vigna species analysed. However, due to the presence of multiple intragenomic 5S types and their incomplete homogenization among V. unguiculata subspecies the relationships in section Catiang could not be resolved below species level. The results presented indicate that intraspecies hybridization might have resulted in the ??horizontal transfer?? of 5S types among the V. unguiculata subspecies, while their maintenance could be due to a slow molecular drive.     

Genetic differentiation of <Emphasis Type="Italic">Vigna</Emphasis> species by RAPD,URP and SSR markers

Seventy genotypes belonging to 7 wild and cultivated Vigna species were genetically differentiated using randomly amplified polymorphic DNA (RAPD), universal rice primer (URP) and simple sequence repeat (SSR) markers. We identified RAPD marker, OPG13 which produced a species-specific fingerprint profile. This primer characterized all the Vigna species uniquely suggesting an insight for their co-evolution, domestication and interspecific relationship. The cluster analysis of combined data set of all the markers resulted in five major groups. Most of the genotypes belonging to cultivated species formed a specific group whereas all the wild species formed a separate cluster using unweighted paired group method with arithmetic averages and principle component analysis. The Mantel matrix correspondence test resulted in a high matrix correlation with best fit (r = 0.95) from combined marker data. Comparison of three-marker systems showed that SSR marker was more efficient in detecting genetic variability among all the Vigna species. The narrow genetic base of the V. radiata cultivars obtained in the present study emphasized that large germplasm collection should be used in Vigna improvement programme.     

Comparison of the epifauna spatial distribution in Posidonia oceanica,Cymodocea nodosa and unvegetated bottoms: Importance of meadow edges

In order to detect the influence of seagrass, the most important habitats of shallow soft-bottom along the Mediterranean coast, on spatial distribution of epifauna, four different types of habitat were sampled: Posidonia oceanica, Cymodocea nodosa, edge of Posidonia meadow and sandy bottoms. Sampling was carried out, using the hand net method, in various random sites and at different times (April 95, August 95 and February 96) on the Alicante coast (SE Spain). A double taxonomic approach was used in order to detect spatial and temporal changes in the abundance of the main taxonomic groups and amphipod species. The differences among habitats were detected by non-parametric multidimensional scaling (MDS) for community structure and by analysis of variance for groups and amphipod populations. The community structure defined by the abundance of taxonomic groups was not significantly modified by the habitats. Single abundance of taxonomic groups also had no significant preference for the type of habitat except in the case of Acari, a group linked to Posidonia. However, certain trends of preference, such as mysids on Posidonia edge and isopods on Cymodocea, were detectable. On other hand, the amphipod assemblage showed important differences among habitats. Some species changed significantly in abundance depending on the habitat considered, e.g. Dexamine spiniventris and Perioculodes longimanus. It was possible to conclude that there is an important influence of Posidonia and Cymodocea on epifauna distribution by diversification of habitat structure on sandy bottoms, but it was more important at the species level than when considering taxonomic groups. Furthermore, sandy bottoms and the meadow edges had a relatively high importance on fauna distribution, depending on taxa and the period of the year.     

Evidences of introgression from cultivated rice to Oryza rufipogon (Poaceae) populations based on SSR fingerprinting: implications for wild rice differentiation and conservation

Crop-to-wild introgression may play an important role in evolution of wild species. Asian cultivated rice (Oryza sativa L.) is of a particular concern because of its cross-compatibility with the wild ancestor, O. rufipogon Griff. The distribution of cultivated rice and O. rufipogon populations is extensively sympatric, particularly in Asia where many wild populations are surrounded by rice fields. Consequently, gene flow from cultivated rice may have a potential to alter genetic composition of wild rice populations in close proximity. In this study, we estimated introgression of cultivated rice with O. rufipogon based on analyses of 139 rice varieties (86 indica and 53 japonica ecotypes) and 336 wild individuals from 11 O. rufipogon populations in China. DNA fingerprinting based on 17 selected rice simple sequence repeat (SSR) primer pairs was adopted to measure allelic frequencies in rice varieties and O. rufipogon samples, and to estimate genetic associations between wild and cultivated rice through cluster analysis. We detected consanguinity of cultivated rice in O. rufipogon populations according to the admixture model of the STRUCTURE program. The analyses showedz that four wild rice populations, DX-P1, DX-P2, GZ-P2, and HL-P, contained some rare alleles that were commonly found in the rice varieties examined. In addition, the four wild rice populations that scattered among the rice varieties in the cluster analysis showed a closer affinity to the cultivars than the other wild populations. This finding supports the contention of substantial gene flow from crop to wild species when these species occur close to each other. The introgressive populations had slightly higher genetic diversity than those that were isolated from rice. Crop-to-wild introgression may have accumulative impacts on genetic variations in wild populations, leading to significant differentiation in wild species. Therefore, effective measure should be taken to avoid considerable introgression from cultivated rice, which may influence the effective in-situ conservation of wild rice species.     

Seed Coat Microsculpturing Is Related to Genomic Components in Wild Brassica juncea and Sinapis arvensis

It has been reported that wild Brassica and related species are widely distributed across Xinjiang, China, and there has been an argument for species identification. Seed coat microsculpturing (SCM) is known to be an excellent character for taxonomic and evolutionary studies. By identifying collections from Xinjiang, China, and combining SCM pattern, flow cytometry, and genome-specific DNA markers as well as sexual compatibility with known species, this study aimed to detect potential relationships between SCM and genomic types in wild Brassica and related species. Three wild collections were found to be tetraploid with a SCM reticulate pattern similar to B. juncea, and containing A and B genome-specific loci, indicating relatively high sexual compatibility with B. juncea. The others were diploid, carrying S-genome-specific DNA markers, and having relatively high sexual compatibility with Sinapis arvensis. Moreover, their SCM was in a rugose pattern similar to that of S. arvensis. It was suggested that SCM, as a morphological characteristic, can reflect genomic type, and be used to distinguish B-genome species such as B. juncea from the related S. arvensis. The relationship between SCM and genomic type can support taxonomic studies of the wild Brassica species and related species.     

Assessment of molecular diversity and evolutionary relationship of kenaf (Hibiscus cannabinus L.), roselle (H. sabdariffa L.) and their wild relatives

Kenaf (Hibiscus cannabinus L.) and roselle (H. sabdariffa L.) are valuable fibre crop species with diverse end use. Phylogenetic relationship of 73 accessions of kenaf, roselle and their wild relatives from 15 countries was assessed using 44 inter-simple sequence repeat (ISSR) and jute (Corchorus olitorius L.) specific simple sequence repeats (SSR) markers. A total of 113 alleles were identified of which 61.95 % were polymorphic. Jute specific SSR markers exhibited high polymorphism and resolving power in kenaf, although ISSR markers exhibited higher resolving power than SSR markers. Number of polymorphic alleles varied from 1 to 5 for ISSR and 1 to 6 for SSR markers. Cultivated species exhibited higher allele polymorphism (57 %) than the wild species (35 %), but the improved cultivars exhibited lower genetic diversity compared to germplasm accessions. Accessions with common genetic lineage and geographical distribution clustered together. Indian kenaf varieties were distinct from cultivars bred in other countries and shared more genetic homology with African accessions. High genetic diversity was observed in the Indian (J = 0.35–0.74) and exotic kenaf germplasm collections (J = 0.38–0.79), suggesting kenaf might have been introduced in India from Africa through Central Asia during early domestication. Genetic similarity-based cluster analysis was in close accordance with taxonomic classification of Hibiscus.     

Study of the relationship between the cultivars of Vitis vinifera and the white-fruited and hermaphrodite Chinese wild grapes

Chinese wild grapes are almost exclusively dioecious and black-fruited, with rare reports of white and hermaphrodite types in V. davidii. To reveal the molecular mechanisms of these phenotypic variations, specific primers were designed to detect the genotypes of mybA-related genes in Vitis species, including the Chinese wild Vitis species, V. riparia, V. rupestris, cultivars of Vitis vinifera and its hybrids. We report here that three mybA-related genes, VvmybA1a, VvmybA2 and VvmybA3, were only detected in cultivars of V. vinifera and its hybrids, but not in V. riparia, V. rupestris or Chinese wild Vitis species, indicating that these genes could be used to test the genetic relationship to V. vinifera. On the other hand, the genes were not detected in the dioecious varieties of V. davidii, but were in the hermaphrodites. In particular, the white-fruited varieties were homozygous for VvmybA1a and showed a low expression of mybA-related genes and UFGT during the entire maturation period. Simple sequence repeat analysis showed that the hermaphrodite varieties of V. davidii, including the white-fruited varieties, were more closely related to V. vinifera cv. Pinot Noir and V. labruscana cv. Kyoho. These results suggested that the white-fruited and hermaphrodite varieties of V. davidii could be the result of its crossing with V. vinifera. It provides a new approach to identify truly Chinese wild varieties and to search for possible hybridization events.     

Mitochondrial gene variation and phylogenetic relationships in the genus Beta

Restriction fragment length polymorphisms (RFLPs) for three mitochondrial genes, coxI, coxII and atpA, were used to determine mitochondrial (mt) DNA diversity in 21 accessions of the genus Beta representing wild and cultivated species. On the basis of distribution of the RFLP patterns these Beta genotypes were assigned into six distinct chondriome groups. A high degree of heterogeneity was found to exist between the mitochondrial genomes of the sugarbeet cultivar and the wild species of Procumbentes section. The polymorphic fragments from wild Beta species were cloned and subjected to fine mapping. We found that most of the RFLPs are due to sequence rearrangements rather than point mutations. Our data also suggest that the close linkage between coxII and coxI is taxonomically localized to an evolutionary lineage that led to Vulgares and Corollinae species but not to Procumbentes species. This linkage is most likely to have arisen via the mutation(s) that inserted the DNA segment containing coxI downstream of coxII in the common ancestor of Vulgares and Corollinae species. The results are discussed with regard to the taxonomic and phylogenetic relationships of the Beta species.     

Inter-primer binding site retrotransposon and inter-simple sequence repeat diversity among wild Lens species

Even though lentil has been an important food legume for centuries, genetic studies in lentil are still in their infancy. Genetic diversity and relationships among wild Lens species from Turkey has seldom been investigated. Additionally, a limited number of simple sequence repeat (SSR) markers have been developed for use in breeding and genetic studies of lentil crop. In this study, molecular characterization of 50 accessions mostly from Turkey, belonging to 6 wild and 1 cultivated Lens species, was performed using newly developed inter-primer binding site (iPBS) retrotransposons and inter-SSR (ISSR) markers. The 10 iPBS primers generated a total of 151 scorable bands, of which 150 were polymorphic (99.3%) with an average of 15.0 polymorphic fragments per primer. The 10 ISSR primers detected 138 scorable bands showing 100% polymorphism, with an average of 13.5 bands per primer. The average polymorphism information content (PIC) value for ISSR markers (0.97) was higher than that for iPBS markers (0.90). Lens orientalis was found to be the most diverse species, raising the possibility of wide crosses with cultivated species Lens culinaris. Cultivated varieties also showed high level of polymorphism, at 82.92% and 51.92% with ISSR and iPBS markers, respectively. Lens lamottei and Lens tomentosus were found as the least polymorphic species using both marker systems. The grouping of accessions and species within clusters were almost similar when iPBS and ISSR graphs were compared. Our data also suggested the role of iPBS-retrotransposons as ‘a universal marker’ for molecular characterization of wild and cultivated Lens species.     

Molecular Diversity in Indian Tobacco Types as Revealed by Randomly Amplified DNA Polymorphism

During the past five decades, a large number of tobacco varieties have been developed for different end uses in India through pure line selection from local land races, mutation breeding, and hybridization involving local selections and exotic introductions followed by pedigree selection. No systematic effort has been made to understand the existing diversity pattern in these varieties, which is crucial to define future breeding strategy in this important commercial crop. We characterized 46 varieties belonging to 10 different manufacturing tobacco types cultivated under different agro-climatic conditions in India along with two wild species of Nicotiana using 40 arbitrary primers in RAPID. The level of polymorphism among the varieties of N. tabacum was 59.4%, which was more than double the level observed in the other cultivated species N. rustica (25.2%). A broader range (0.64 to 0.94) of pair wise similarity measures in N. tabacum than in N. rustica (0.83 to 0.92) reflected the more diversified breeding efforts in the major cultivated species. The two wild species namely, N. glutinosa and N. gossei clustered separately from the two cultivated species. Molecular classification of the varieties corresponded largely with their manufacturing trait and parentage. RAPID markers provided sufficient resolution to distinguish among closely related tobacco types. Nine RAPID markers were found conserved across all the varieties and species. The markers found specific to the varieties can be used in correct identification of the carrier genotypes in trade and commerce. This is the first report on the molecular diversity analysis of Indian tobacco.     

Morphometric analysis of varieties of Acacia caven: (Leguminosae, Mimosoideae): Taxonomic inferences in the context of other Argentinean species

The aims of this study were to evaluate the degree of morphological differentiation between six varieties of Acacia caven and to examine their taxonomic validity in the context of other Argentinean species of the genus. To accomplish these purposes, morphological traits have been analyzed using multivariate methods (non parametric ANOVA, phenetic analysis and principal component analysis) on the varieties of A. caven and other six species of the genus, represented in Argentina. The phenogram obtained showed two principal clusters, one grouping all the species of subg. Acacia and the other grouping the species of subg. Aculeiferum. This result agreed with Vassal's infrageneric classification. However, the results of the principal PCA gathered the seven species here included in three groups, which were consistent with Bentham's infrageneric treatment. The ANOVA method indicated that most of the morphometric characters used were statistically sound for differentiation between varieties of A. caven. Further studies, including more species and characters, must be performed in order to clarify the position of Acacia boliviana and the relationships between A. caven and A. curvifructa.     

Comparative phytochemistry of eleven species of Vigna (Fabaceae)

A survey of the biochemical constituents of 11 species of Vigna indicates the absence of the non-protein amino acid canavanine in their seeds, and absence of proanthocyanidin (polyphenol) in their leaves. Proanthocyanidin was found in the seeds of all, except Vigna subterranea. The constitutive leaf flavonoids of four genotypes of the pantropic V. subterranea were also studied and compared with those from three other cultivated species. The flavonoid kaempferol seems to be most prevalent as it was found in all of the four cultivated species and genotypes. The glycoside kaempferol-3-O-rutinoside was found present in the four genotypes of V. subterranea and other cultivated Vigna species. However, the flavonoid kaempferol-3-O-glucoside-7-rhamnoside is restricted to V. subterranea. This study questions the inclusion of V. subterranea in the genus Vigna on account of absence of seed proanthocyanidin and restricted accumulation of kaempferol-3-O-glucoside-7-rhamnoside in the leaves.