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.     

Purification and characterization of a Bowman-Birk proteinase inhibitor from the seeds of black gram (Vigna mungo)

A proteinase inhibitor (BgPI) was purified from black gram, Vigna mungo (cv. TAU-1) seeds by using ammonium sulfate fractionation, followed by ion-exchange, affinity and gel-filtration chromatography. BgPI showed a single band in SDS-PAGE under non-reducing condition with an apparent molecular mass of ∼8 kDa correlating to the peak 8041.5 Da in matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrum. BgPI existed in different isoinhibitor forms with pI values ranging from 4.3 to 6.0. The internal sequence “SIPPQCHCADIR” of a peak 1453.7 m/z, obtained from MALDI-TOF-TOF showed 100% similarity with Bowman-Birk inhibitor (BBI) family. BgPI exhibited non-competitive-type inhibitory activity against both bovine pancreatic trypsin (K_i of 309.8 nM) and chymotrypsin (K_i of 10.7 μM), however, with a molar ratio of 1:2 with trypsin. BgPI was stable up to a temperature of 80 °C and active over a wide pH range between 2 and 12. The temperature-induced conformational changes in secondary structure are reversed when BgPI was cooled from 90 to 25 °C. Further, upon reduction with dithiothreitol, BgPI lost both its inhibitory activity as well as secondary structural conformation. Lysine residue(s) present in the reactive site of BgPI play an important role in inhibiting the bovine trypsin activity. The present study provides detailed biochemical characteristic features of a BBI type serine proteinase inhibitor isolated from V. mungo.     

Flavonoids of someVigna-plants in leguminosae

Flavonoids in five lines ofVigna mungo, fifteen lines ofV. radiata var.radiata and two lines ofV. radiata var.sublobata, which belong to the subgenus Ceratotropis, were examined. In hypocotyls, seed-coats and mature leaves of those plants, twelve kinds of flavonoid including three anthocyanins, two leucoanthocyanins, two glycoflavones and five flavonol glycosides were found, and their distribution pattern in twenty-two lines of the legumes is discussed. The leaves ofV. radiata var.radiata and var.sublobata contained the glycosides (mainly rutin) of both quercetin and kaempferol, while those ofV. mungo contained only kaempferol glycosides, with robinin as the predominant pigment, and the purple-red hypocotyls of the former group contained delphinidin 3-p-coumaroylglucoside, while those of the latter contained cyanidin 3-glucoside, although delphinidin 3-glucoside was commonly found in all of the plants. With the exception of two lines, all of the seed-coats examined contained in common four compounds of vitexin, isovitexin, leucocyanidin and leucodelphinidin, whereas in addition to these pigments the black seed-coats ofV. mungo andV. radiata var.sublobata TC 1965 contained delphindin 3-glucoside.     

Growth and metal accumulation potential of Vigna radiata L. grown under fly-ash amendments

The study was undertaken with the aim of testing the feasibility of growing Vigna radiata on fly-ash dykes or on fly-ash contaminated soil. Experiments were conducted in earthen pots (12″) having various amendments of fly-ash with garden soil, press mud and farm yard manure and harvested after 60 days of growth. Various growth variables were analyzed, viz., root-shoot length, biomass yield, number of branches, photosynthetic area and pigments, protein content, in vivo nitrate reductase activity and nodulation frequency. The metal accumulation potential (Fe, Zn, Cu, Cr, Cd) in roots and shoot tissues of plants was determined in various fly-ash amendments and seeds were also examined for their metal content so that they can be utilized for growing in fly-ash-contaminated areas. Our results show that the plants grown in 100% fly-ash accumulated significant amounts of metals due to which general vigor and development of plant are affected. V. radiata grown on ameliorated fly-ash showed tolerant characteristics. Plants accumulated metal in the order Fe > Cu > Zn > Cr > Cd. However, their concentration was more in shoot than root tissue except in the case of Fe, which was found to have accumulated more in root tissues. Press mud (25%) amended with fly-ash was found to be the best ameliorant to support plant growth. It may be concluded from the present study that V. radiata was an ideal crop for growing in fly-ash-contaminated areas for the revegetation of fly-ash landfills.     

Ontogeny of Somatic Embryos of Vigna aconitifolia, Vigna mungo and Vigna radiata

Callus cultures were initiated from immature cotyledons of Vignaaconitifolia, V. mungo and V. radiata on MS medium supplementedwith NAA, picloram or 2, 4-D. On transfer to L-6 liquid mediumsupplemented with low concentrations of picloram, GA₃ and cytokinins,large number of somatic embryos differentiated from the callus.The cells destined to become somatic embryos divided to formspherical or filamentous proembryos. From the filamentous proembryo,the embryo proper developed either at single or multiple sites.Development of somatic embryos from multiple sites resultedin several embryos connected by a common suspensor at the radicleend. Continued divisions of the proembryos led to globular,heart shaped and dicotyledonary stages of somatic embryogenesis.The somatic embryos of V. mungo and V. aconitifolia differentiatedinto tiny plantlets at low frequency (1%) in liquid suspensioncultures supplemented with zeatin, picloram and GA₃. Vigna aconitifolia Jacq, Marechal, mothbean, Vigna mungo L. Hepper, urdbean, Vigna radiata L. Wilczk, mungbean, somatic embryo     

Vigna mungo, V. radiata and V. unguiculata plants sampled in different agronomical–ecological–climatic regions of India are nodulated by Bradyrhizobium yuanmingense

Vigna mungo, Vigna radiata and Vigna unguiculata are important legume crops cultivated in India, but little is known about the genetic resources in native rhizobia that nodulate these species. To identify these bacteria, a core collection of 76 slow-growing isolates was built from root nodules of V. mungo, V. radiata and V. unguiculata plants grown at different sites within three agro-ecological-climatic regions of India. The genetic diversity of the bacterial collection was assessed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified DNA fragments of the 16S–23S rDNA intergenic spacer (IGS) region, and the symbiotic genes nifH and nodC. One rDNA IGS type grouped 91% of isolates, but more diversity was found at the symbiotic loci (17 symbiotic genotypes). Overall, no host plant specificity was shown, the three host plant species sharing common bradyrhizobial genotypes that represented 62% of the collection. Similarly, the predominant genotypes were found at most sampling sites and in all agro-ecological-climatic regions. Phylogenies inferred from IGS sequencing and multi-locus sequence analysis of the dnaK, glnII and recA genes indicated that all isolates but one were clustered with the Bradyrhizobium yuanmingense species. The nifH phylogeny also grouped the different nif haplotypes within a cluster including B. yuanmingense, except for one infrequent nif haplotype which formed a new lineage within the Bradyrhizobium genus. These results may reflect a long history of co-evolution between B. yuanmingense and Vigna spp. in India, while intra-species polymorphism detected in the symbiotic loci may be linked with the long history of diversification of B. yuanmingense coinciding with that of its host legumes.     

Seed protein and amino acid composition of WildVigna radiata var.sublobata (Fabaceae) and Two Cultigens,V. mungo andV. radiata

WildVigna radiata var.sublobata populations, inhabiting the mountains of India, and two cultigens,V. mungo andV. radiata, were examined for seed-protein content and amino acid composition. The protein content in the populations varied from 15.2 to 21%. Lysine, valine, isoleucine, leucine, phenylalanine, and tyrosine contents were invariably higher in wild populations as compared with the FAO reference pattern. The variation encountered in wild populations is largely genetic in origin. Total essential amino acids in the wild populations varied from 38.3 to 42.2; in the cultigens, from 29.2 to 37.5 (g/100 g protein). There was marked variation in the amino acid composition of the populations for the two different ecozones studied, suggesting a broad genetic base.     

Molecular Marker-Assisted Genotyping of Mungbean Yellow Mosaic India Virus Resistant Germplasms of Mungbean and Urdbean

Mungbean Yellow Mosaic India Virus (MYMIV) belonging to the genus begomovirus causes the yellow mosaic disease in a number of economically important edible grain legumes including mungbean (Vigna radiata), urdbean (Vigna mungo) and soybean (Glycine max). The disease is severe, critical, open spread and inflicts heavy yield losses annually. The objective of this study is to develop molecular markers linked to MYMIV-resistance to facilitate genotyping of urdbean and mungbean germplasms for MYMIV-reaction. Resistance-linked molecular markers were successfully developed from consensus motifs of other resistance (R) gene or R gene homologue sequences. Applying linked marker-assisted genotyping, plant breeders can carry out repeated genotyping throughout the growing season in absence of any disease incidence. Two MYMIV-resistance marker loci, YR4 and CYR1, were identified and of these two CYR1 is completely linked with MYMIV-resistant germplasms and co-segregating with MYMIV-resistant F₂, F₃ progenies of urdbean. The present study demonstrated that these two markers could be efficiently employed together in a multiplex-PCR-reaction for genotyping both V. mungo and V. radiata germplasms from field grown plants and also directly from the seed stock. This method of genotyping would save time and labour during the introgression of MYMIV-resistance through molecular breeding, as methods of phenotyping against begomoviruses are tedious, labour and time intensive.     

In vitro induction of multiple shoots and plant regeneration inVigna

Summary Cotyledonary nodes, excised cotyledons, and hypocotyl segments of six varieties ofVigna mungo andV. radiata have been tested for their morphogenic potential on media containing a range of hormonal combinations including benzyladenine, kinetin, thidiazuron (TDZ), and zeatin. Multiple shoots developed on cotyledonary node explants in all varieties tested on basal medium containing cytokinin. Presence of both the cotyledons, either full or half, resulted in a maximum number of shoots produced. Shoot bud regeneration was achieved via meristem formation on excised cotyledons on Murashige-skoog basal medium with B₅ vitamins supplemented with TDZ. Mature plants had normal phenotypes.V. mungo var. PS1 andV. radiata var. Pusa 105 were found to be the most responsive varieties for shoot regneration. The histology ofin vitro organogenesis was studied.     

Vigna unguiculata is nodulated in Spain by endosymbionts of Genisteae legumes and by a new symbiovar (vignae) of the genus Bradyrhizobium

Vigna unguiculata was introduced into Europe from its distribution centre in Africa, and it is currently being cultivated in Mediterranean regions with adequate edapho-climatic conditions where the slow growing rhizobia nodulating this legume have not yet been studied. Previous studies based on rrs gene and ITS region analyses have shown that Bradyrhizobium yuanmingense and B. elkanii nodulated V. unguiculata in Africa, but these two species were not found in this study. Using the same phylogenetic markers it was shown that V. unguiculata, a legume from the tribe Phaseolae, was nodulated in Spain by two species of group I, B. cytisi and B. canariense, which are common endosymbionts of Genisteae in both Europe and Africa. These species have not been found to date in V. unguiculata nodules in its African distribution centres. All strains from Bradyrhizobium group I isolated in Spain belonged to the symbiovar genistearum, which is found at present only in Genisteae legumes in both Africa and Europe. V. unguiculata was also nodulated in Spain by a strain from Bradyrhizobium group II that belonged to a novel symbiovar (vignae). Some African V. unguiculata-nodulating strains also belonged to this proposed new symbiovar.     

Impacts of ambient solar uv (280-400 nm) radiation on three tropical legumes

Tropical regions receive the highest level of global solar ultraviolet (UV) radiation especially UV-B (280-320 nm). The average daily dose of the UV-B radiation in Madurai, South India (10°N) is 10 kJ m^-2. This is approximately 50% more than the average daily UV-B radiation in many European countries. A field study was conducted using selective filters to remove either the UV-B (< 320 nm) or UV-B/A (<400 nm) of the solar spectrum, and the effects were followed inCyamopsis tetragonoloba, Vigna mungo, andVigna radiata to determine their sensitivity to UV. When compared to ambient radiation, exclusion of solar UV-B increased the seedling height, leaf area, fresh weight and dry weight and the crop yield by 50% in the case ofCyamopsis, and the extent of such increase was slightly less under UV-B/A exclusion. InV. mungo a significant reduction was seen in solar UV excluded plants whileV. radiata was found to be unaffected.     

Etude chimiotaxonomique dans les genres Macrotyloma, Dolichos et Pseudovigna

The variations of free, protein and total amino acid contents have been studied in seeds of Macrotyloma, Dolichos and Pseudovigna. The concentrations of free amino acids and of γ-glutamylphenylalanine seem to be characteristic of some species and also of some genera.     

Dipeptide precursor of garlic odour in Marasmius species

γ-Glutamyl-marasmine, a new natural dipeptide containing an unusual cysteine sulphoxide moiety has been isolated from the Basidiomyceteous mushrooms Marasmius alliaceus, M. scorodonius and M. prasiosmus, which are known for their garlic like odour. It is shown that this compound is the common natural precursor and that its two step enzymatic cleavage leads to the odorous substances. In the first step γ-glutamyl -marasmine is cleaved by a γ-glutamyl transpeptidase. The formed marasmine is split in a second enzymatic reaction by a C-S lyase into pyruvic acid, ammonia and an unstable sulfur compound, which decomposes to form the odorous secondary products.     

Vigna radiata var.Sublobata (Fabaceae): Economically useful wild relative of urd and mung beans

Eight natural populations ofVigna radiata var.sublobata— wild relative of cultivated urd (V. mungoj and mung (V. radiata,) beans—were sampled from different ecozones of Palney Hills, an eastward offshoot of Western Ghats of Tamilnadu, India. Photosynthetic efficiency, protein content, seed weight, and amino acid composition were determined for it and the cultigens. Some populations ofV. radiata var.sublobata are as good as or even superior to the cultigens. The wild relative is a potential donor of desirable traits to urd and mung beans.     

Sequence related amplified polymorphism (SRAP) analysis for genetic diversity and micronutrient content among gene pools in mungbean [Vigna radiata (L.) Wilczek]

Vigna radiata (L.) Wilczek, commonly called mungbean is an important pulse crop. Commercial cultivars contain low levels of iron and zinc and it is important to assess genetic variability in the available germplasm for improving micronutrient content in commercial cultivars. The present study was undertaken to study molecular diversity using Sequence-related amplified polymorphism (SRAP) among 21 Vigna radiata genotypes. Twenty nine SRAP primer combinations produced a total of 121 amplified bands which were polymorphic with an average of 4.65 bands per primer. The size of amplified bands ranged from 70 bp to 3,000 bp and 6 out of 29 SRAP primers were most useful in fingerprinting Vigna radiata genotypes under study. The similarity coefficients between different genotypes ranged from 0.45 to 0.96 with an average similarity value of 0.71. At an arbitrary cut-off at 60 % similarity level on a dendrogram, the Vigna radiata accessions were categorized into two major clusters. ML1108 and 2KM115 were found to be genetically similar. SMH99-1A and ML776 showed high iron and zinc content while Satya was poor in iron as well as zinc content. Mapping population involving ML776 and Satya could be used for tagging gene(s) for micronutrient content. The results indicated that SRAP markers were efficient for identification of Vigna radiata genotypes and assessment of the genetic relationships among them.     

Un nouvel acide amine dans les graines de Vigna vexillata

A new natural amino acid, l-(+)-p-aminophenylalanine, has been isolated from a legume, Vigna vexillata (L.) A. Rich. Its structure was determined and confirmed by synthesis.     

Detoxification potential and expression analysis of eutypine reducing aldehyde reductase (VrALR) during progressive drought and recovery in Vigna radiata (L.) Wilczek roots

Generation of reactive oxygen species (ROS) in plants is an inevitable consequence of adverse environmental cues and the ability to detoxify deleterious by-products of ROS-mediated oxidation reactions reflect an important defence strategy to combat abiotic stress. Here, we have cloned the eutypine reducing aldehyde reductase gene (VrALR) from Vigna radiata (L.) Wilczek roots. We have expressed and purified the VrALR protein and analyzed its enzyme kinetic parameters and catalytic efficiency with three different substrates to confirm its identity. The functional characterization of this enzyme was unravelled through heterologous expression of the gene in Escherichia coli BL21 and an oxidative stress-sensitive Saccharomyces cerevisiae mutant strain, W3O3-1-A. Finally, the endogenous VrALR enzyme activity and the mRNA expression patterns of the VrALR gene in the roots of V. radiata in response to progressive drought stress in vivo was studied to correlate the ROS-detoxifying role of this important enzyme under the influence of progressive drought stress. Our results, for the first time, demonstrate that eutypine reducing VrALR provides varying degree of stress tolerance in bacteria, yeast systems and also plays a promising protective role against oxidative stress in V. radiata roots during gradual water deprivation. The present study provides an unequivocal evidence to understand the crucial role of aldehyde reductase ROS-detoxifying system which is highly essential for developing stress tolerance in economically important crop plants.     

Distribution of low molecular weight carbohydrates in the subgenusCeratotropis of the genusVigna (Leguminosae)

The seeds of 9 members of the subgenusCeratotropis (Piper) Verdc., namelyVigna aconitifolia (Jacq.) Maréchal,V. angularis (Willd.) Ohwi et Ohashi,V. minima (Roxb.) Ohwi et Ohashi,V. nakashimae (Ohwi) Ohwi et Ohashi,V. reflexo-pilosa Hayata,V. umbellata (Thumb.) Ohwi et Ohashi,V. mungo (L.) Hepper,V. radiata (L.) Wilczek andV. sp., have been examined. On their low molecular weight carbohydrate compositions, this subgenus has been divided into 2 subgroups, mungo-radiata group and angularis group. Four other species referred to the subgeneraPlectotropis (Schumach.) Bak.,Lasiospron (Benth. emend Piper) Maréchal, Mascherpa et Stainier andVigna, V. vexillata (L.) A. Rich.,V. lasiocarpa (Benth.) Verdc.,V. marina (Burm.) Merr. andV. unguiculata (L.) Walp., were also analyzed and they had distinctive carbohydrate compositions. 1d-4-O-methyl-myo-inositol and 1d-5-O-(α-d-galactopyranosyl)-4-O-methyl-myo-inositol were detected in all species examined exceptV. vexillata, V. mungo andV. radiata.     

Induction of stress metabolite formation in suspension cultures of Vigna angularis

Three isoflavone glucosides were isolated from cell suspension cultures of red bean (Vigna angularis) treated with actinomycin D and identified as