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PubMed2004

Analysis of an isolate of Mungbean yellow mosaic virus (MYMV) with a highly variable DNA B component.

One DNA A (KA30) and five different DNA B components (KA21, KA22, KA27, KA28 and KA34) of a geminivirus, Mungbean yellow mosaic virus-Vigna (MYMV-Vig) were cloned from a pooled sample of field-infected Vigna mungo plants from Vamban, South India. MYMV-Vig DNA A (KA30) and one of the DNA B components (KA27) exhibited 97% and 95% sequence identities, respectively, to those of MYMV reported from Thailand. However, the DNA B components KA21, KA22, KA28 and KA34 exhibited only 71 to 72% sequence identity to MYMV DNA B. Co-existence of multiple DNA B components in field-infected V. mungo was proved by Southern and PCR analyses. Each of the five DNA B components was infective together with the DNA A upon agroinoculation. Agroinoculation with mixed cultures of Agrobacterium with partial dimers of DNA A and all five DNA Bs proved that all five DNA B components can co-infect a single V. mungo plant.

Blotting, Southern↗
PubMed1980

Cell-free synthesis of active ribulose-1,5-bisphosphate carboxylase in the mesophyll chloroplasts of Sorghum vulgare.

Chloroplast and whole leaf cell RNA from Vigna sinensis, a C3 plant were used as exogenous templates for translation in a cell-free light-dependent system of isolated chloroplasts from Sorghum vulgare, a C4-type plant. Analysis of immunoprecipitates of the translated products with the total cellular RNA on sodium dodecyl sulfate polyacrylamide gels revealed the synthesis of both the subunits of ribulose-1,5-bisphosphate carboxylase. Similar analysis of the product translated with the RNA from Vigna chloroplasts, indicated the synthesis of only the large subunit of the carboxylase. Apparently the chloroplast protein synthetic machinery is capable of translating the mRNA for the smaller subunit of this protein as well, which is known to be translated in the cytoplasmic ribosomal system. Sufficient quantities of ribulose-1,5-bisphosphate carboxylase were synthesised in vitro in the preincubated chloroplast system with the whole cell RNA from the C3 plant to assay the ribulose 1,5-bisphosphate-dependent carboxylation. The newly synthesised protein in the cell-free system is identical in many ways to the native enzyme including the Mg2+ concentration-dependent shift in pH optima towards neutral side. It is specifically inhibited by anti-native ribulose-1,5-bisphosphate carboxylase and pyridoxal 5'-phosphate.

Carboxy-Lyases↗
PubMed1991

Sensitivity of three leguminous crops to O3 as influenced by different stages of growth and development.

The sensitivity of Cicer arietinum, Vigna mungo and Trigonella foenum-graecum to O(3) has been assessed at different stages of growth and development. Plants of different ages (0, 1, 2, 4 and 6 weeks old) were fumigated with 0 and 120 nl litre(-1) O(3), from 09.30 h to 16.30 h each day for four weeks, in hemispherical chambers located out-of-doors. Seed germination was not affected by O(3) in any of the species, but there were responses (differing between species) on the cotyledons. True leaves were fairly resistant when young but later they became more sensitive. Premature senescence and earlier abscission of leaves (in C. arietinum and T. foenum-graecum) and flowers and abortive fruit drop (in C. arietinum) were also observed. Of the five growth stages examined, 2- and 4-week-old plants seemed to be most sensitive except for Trigonella where sensitivity decreased with increasing age of the plants. The partitioning and distribution of dry matter among different plant parts was also significantly disturbed and root, leaf and stem were adversely affected in a decreasing order. However, the percentage reductions in dry weight per plants for Cicer and Vigna increased with age up to four weeks, then declined abruptly. Growth reductions at the 0- and 6-week-old stages differed only slightly and were very small in magnitude. It may, therefore, be suggested that the plants of these legumes in early stages of exponential growth are more vulnerable to O(3) damage and that the developmental or physiological age is an important factor in O(3) sensitivity.

Journal Article↗
PubMed1975

Biosynthesis of p-aminophenylalanine: part of a general scheme for the biosynthesis of chorisimic acid derivatives.

p-Aminophenylalanine is biosynthesized in Vigna vexillata (L.) A. Rich. from shikimic acid through a pathway different from that giving phenylalanine and tyrosine. Experiments with 1,6-14C-labelled shikimic acid demonstrate that the C3-side chain in p-aminophenylalanine is attached to the original C-1 in shikimic acid. The biosynthesis of p-aminophenylalanine in Vigna vexillata probably follows the same pathway as the biosynthesis of this amino acid in Streptomyces species where it is known to be an intermediate in the biosynthesis of chloramphenicol. It is proposed that the biosynthesis takes place through chorismic acid, 4-amino-3-enolpyruvylcyclohexa-1,5-dienecarboxylic acid, 3-(4-amino-1-carboxycyclohexa-2,5-dienyl) pyruvic acid, and 4'-aminophenyl-yruvic acid. It is proposed that chorismic acid can gave rise to 4-amino-3-enolpyruvylcyclohexa-1,5-dienecarboxylic acid, 2-amino-3-enolpyruvylcyclohexa-4,6-dienecarboxylic acid, and isochorismic acid, and that these three compounds and chorismic acid itself by simple rearrangements and elimination reactions can give rise to most known chorismic acid derivatives, i. e.p-hydroxybenzoic acid, 3,4-dihydroxy-3,4-dihydrobenzoic acid, phenylalanine, tyrosine, p-aminobenzoic acid, p-aminophenylalanine, anthranilic acid, 2-amino-3-hydroxy-2,3-dihydrobenzoic acid, 3-(3-carobxyphenyl) alanine, 3-(3-carbocy-4-hydrocyphenyl) alanine, salicylic acid, and 2,3-dihydroxy2,3-dihydrobenzoic acid.

Amines↗
PubMed2006

Cloning and characterization of a cowpea seed lipid transfer protein cDNA: expression analysis during seed development and under fungal and cold stresses in seedlings' tissues.

Lipid transfer proteins (LTPs) are antimicrobial peptides (AMPs) involved in the defense of plants against pathogens. Our group has previously characterized and purified a LTP from cowpea (Vigna unguiculata (L.) Walp.) seeds which caused the inhibition of growth of fungal pathogens in vitro. The aim of this work was to obtain the cDNA encoding the cowpea LTP and after cloning, to use the cDNA as a probe for studying its expression profile during the development of cowpea seeds. In this work, the N-terminal sequence of the mature LTP peptide from cowpea was used to produce a degenerated oligonucleotide. This primer allowed the amplification of the LTP cDNA by RT-PCR from mRNA of cowpea seeds. The sequence analysis of the cloned cDNA, named VULTP, showed 494 bp which encoded a polypeptide of 91 amino acids. The deduced peptide presented high homology of similarity to plant LTPs of Vigna radiata var. radiate (94%), Prunus domestica (82%) and Zea mays (72%). The expression profile of the VULTP gene in cowpea was analyzed by Northern blot and revealed that the transcript is not accumulated in adult tissues. Conversely, VULTP mRNA is early and strongly accumulated during seed development. The results obtained to seedling of cowpea demonstrate that the VULTP gene presents differential expression in response to different stress. Further studies will be conducted to try to gain better understanding about the physiological role of this gene in cowpea.

Adaptation, Physiological↗
PubMed2000

A rapid and sensitive assay method for measuring amine oxidase based on hydrogen peroxide-titanium complex formation.

Hydrogenperoxide (H(2)O(2)) is an end product of diamine and polyamine oxidation by their respective oxidase enzymes. A new sensitive assay method is based on a H(2)O(2)-titanium (Ti) complex formation as an indicator of H(2)O(2) production due to polyamine oxidation. The orange-yellow coloured H(2)O(2)-Ti complex was measured at 410 nm in a Shimadzu spectrophotometer. The assay conditions for maximum diamine oxidase (DAO) and polyamine oxidase (PAO) as standardized here using the hypocotyl tissues of Vigna catjang Endl. cv Pusa Barsati consisted of pH 7.4 (40 mM potassium phosphate buffer), 3 mM substrate (putrescine or spermine), 37 degrees C incubation temperature and 30 min incubation time in the presence of catechol (10(-2) M) used as an inhibitor of both peroxidase and catalase activity. The method described here was significantly more sensitive than the starch-iodide method [T.A. Smith, Biochem. Biophys. Res. Commun. 41 (1970) 1452-1456], which could be improved further if measured under the same assay conditions as described for the H(2)O(2)-Ti method. Sensitivity of the present method was tested by assaying DAO/PAO activity in auxin treated hypocotyls of Vigna and comparing it with the starch-iodide method in two other plant samples.

Journal Article↗
PubMed1998

Legume vicilins (7S storage globulins) inhibit yeast growth and glucose stimulated acidification of the medium by yeast cells.

Vicilin (7S storage proteins) isolated from different legume seeds were shown to inhibit yeast growth and glucose stimulated acidification of the medium by yeast cells. The degree of growth inhibition varied with the origin of vicilins. It was more than 90% for vicilins from cowpea (Vigna unguiculata, cultivar pitiuba) and equal to 65% for vicilins from Vigna radiata, in the case of Saccharomyces cerevisae. Vicilins from cowpea seeds inhibited the glucose stimulated acidification of the medium by S. cerevisae up to 60%. We have also observed that vicilins bind to yeast cells. We suggest that vicilins bind to chitin-containing structures of yeast cells and that such association could result in inhibition of H+ pumping, cell growth and spore formation. A final consequence of the yeast growth inhibition by vicilins is (probably) the formation of spores.

Antifungal Agents↗
PubMed1998

Analysis of the expression of two thiolprotease genes from daylily (Hemerocallis spp.) during flower senescence.

A cDNA clone encoding a daylily (Hemerocallis spp.) thiolprotease (SEN11), whose expression is strongly upregulated in flower tepal senescence, has been isolated. The amino acid sequence, deduced from the nucleotide sequence, showed highest similarity to plant thiolproteases of Vigna mungo, Phaseolus vulgaris and Hemerocallis (SEN102), and contains a putative ER retention signal that has been described in Vigna mungo. SEN102 and SEN11 transcripts were not detectable in flower buds at the opening stage, but two peaks of transcripts were seen after 9 h and 19 h, in both petals and sepals, when wilting symptoms were apparent. The pattern of protease activity migrating on a 26.3 kDa protein was similar to the SEN102 and SEN11 transcript profiles. These two genes were also expressed in stamens and leaves, but their transcripts were undetectable in carpels and rhizomes. The expression of SEN102 was lower in the senescent leaf than in the green leaf. The pattern of expression of these genes suggests their involvement in the protein hydrolysis occurring in tepals at the late senescence stage, whereas in leaves they could be involved in the constitutive protein turnover machinery. Exogenous gibberellic acid application to cut flowers increased transcripts of both genes.

Amino Acid Sequence↗
PubMed2001

Cloning and characterization of a DEAD box RNA helicase from the viable seedlings of aged mung bean.

Seeds stored under adverse conditions will reduce the viability of germination as a result of induced aging. We have established a procedure to induce accelerated aging for studying the process of aging in mung bean (Vigna radiata) seeds at the molecular level. A full-length cDNA was isolated from acceleratedly aged mung bean seedlings. The cDNA, VrRH1 (Vigna radiata RNA helicase 1), contains an open reading frame of 2139 bp encoding a protein of 713 amino acids. VrRHI has seven highly conserved motifs including the DEAD box as in the case of other plant RNA helicases. VrRHI was sub-cloned into an expression vector pET-28b (+), over-expressed in Escherichia coli BL 21 and purified by a Ni2+-agarose column. The expressed protein showed double-stranded RNA unwinding and ATPase activities. Either ATP or dATP is required for the unwinding activity, indicating that VrRHI is an ATP/dATP-dependent RNA helicase. Northern blot analysis showed the presence of mRNAs hybridized with a full-length cDNA fragment of VrRHI (VrRH transcripts) in mung bean seeds that were imbibed for 16 to 32 h after accelerated aging treatment. The amount of these mRNAs reached a maximum in 24 h imbibed seeds after the treatment. The accumulation of VrRH transcripts was shown to lead to the appearance of 25S and 18S rRNAs in the imbibed aging mung bean seeds. The results suggest that VrRHI may play a role in the viability of mung bean seeds.

Adenosine Triphosphatases↗
PubMed1990

Proposed regulatory pathway encoded by the nodV and nodW genes, determinants of host specificity in Bradyrhizobium japonicum.

Bradyrhizobium japonicum is the root nodule endosymbiont of soybean (Glycine max), mung bean (Vigna radiata), cowpea (Vigna unguiculata), and Siratro (Macroptilium atropurpureum). We report the characteristics of a nodulation-gene region of B. japonicum that contributes only marginally to the bacterium's ability to nodulate soybean but is essential for the nodulation of the three alternative hosts. This DNA region consists of two open reading frames designated nodV and nodW. The predicted amino acid sequences of the NodV and NodW proteins suggest that they are members of the family of two-component regulatory systems, which supports the hypothesis that NodV responds to an environmental stimulus and, after signal transduction, NodW may be required to positively regulate the transcription of one or several unknown genes involved in the nodulation process. It seems likely that all host plants produce the necessary signal, whereas host specificity may be brought about by the product(s) of the gene(s) activated by NodW.

Amino Acid Sequence↗
PubMed1997

Purification and characterization of two novel hypersensitive response-inducing specific elicitors produced by the cowpea rust fungus.

Two novel elicitor peptides that are produced by the race 1 of the cowpea rust fungus Uromyces vignae and that specifically induce a hypersensitive response (a putative form of programmed cell death) in a resistant cultivar of cowpea (Vigna unguiculata L. Walp) have been purified to homogeneity. Purification steps included gel filtration, anion-exchange chromatography, continuous elution electrophoresis, and reversed-phase C18 high performance liquid chromatography. The relative molecular masses of the peptide elicitors as deduced from Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis were 5600 Da (major) and 5800 Da (minor), respectively. Peptide 1 (major) and the minor copurifying peptide (peptide 2) resolved at the final C18 high performance liquid chromatography step. The NH2 terminus of peptide 1 was deblocked with anhydrous trifluoroacetic acid prior to sequencing. However, the NH2 terminus of peptide 2 was free. The acidic and hydrophobic peptides show some homology between themselves but do not show any significant similarity with known proteins. The two specific elicitors may be products of two avirulence genes corresponding to the two genes for resistance in the resistant cultivar.

Amino Acid Sequence↗
PubMed2003

Chemical composition of certain tribal pulses in South India.

Raw seeds of tribal pulses Atylosia scarabaeoides, Canavalia gladiata, Lablab purpureus var. lignosus, Neonotonia wightii var. coimbatorensis, Rhynchosia filipes, Vigna trilobata and Vigna unguiculata subsp. unguiculata were investigated for their proximate composition, minerals, vitamins (niacin and ascorbic acid) and certain anti-nutritional substances. The seeds of L. purpureus var. lignosus and V. trilobata had a higher content of crude protein than the commonly consumed Indian pulses. The seeds were found to be a rich source of minerals like potassium when compared with recommended dietary allowance values. The total free phenolics, tannins, 3,4-dihydroxyphenylalanine and hydrogen cyanide were also analysed.

Developing Countries↗
PubMed1992

Evidence for orthologous seed weight genes in cowpea and mung bean based on RFLP mapping.

A well saturated genomic map is a necessity for a breeding program based on marker assisted selection. To this end, we are developing genomic maps for cowpea (Vigna unguiculata 2N = 22) and mung bean (Vigna radiata 2N = 22) based on restriction fragment length polymorphism (RFLP) markers. Using these maps, we have located major quantitative trait loci (QTLs) for seed weight in both species. Two unlinked genomic regions in cowpea contained QTLs accounting for 52.7% of the variation for seed weight. In mung bean there were four unlinked genomic regions accounting for 49.7% of the variation for seed weight. In both cowpea and mung bean the genomic region with the greatest effect on seed weight spanned the same RFLP markers in the same linkage order. This suggests that the QTLs in this genomic region have remained conserved through evolution. This inference is supported by the observation that a significant interaction (i.e., epistasis) was detected between the QTL(s) in the conserved region and an unlinked RFLP marker locus in both species.

Fabaceae↗
PubMed1982

Novel evolutionary variation in transcription and location of two chloroplast genes.

We have found major evolutionary changes in the types of transcripts produced by specific chloroplast genes, in particular those encoding the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase and a photosystem II polypeptide (PII). Two distinct patterns of LS gene transcripts are revealed by hybridizing an LS gene probe to electrophoretically separated RNA from 19 angiosperms. Most species, including pea, contain the single transcript of approximately 1.6 kb previously observed in corn, spinach and mustard. However, in mung bean and other members of the legume genera Vigna and Phaseolus, the 1.6 kb transcript represents only a minor fraction of LS transcripts, and instead, two larger LS transcripts of approximately 2.4 and 2.6 kb predominate. The PII gene produces a single transcript in pea and most other species examined, while members of the related legume genera Vigna, Phaseolus and Glycine contain two additional transcripts which are smaller in size and probably represent specific RNA breakdown products. A single species, sweet pea (Lathyrus odoratus), contains a second PII transcript which is 0.2 kb larger than the approximately 1.2 kb transcript found in all species. The LS and PII genes map to the same 5 kb region in both pea and mung bean and are transcribed off the same DNA strand. In contrast, published studies indicate that the two genes are approximately 50 kb apart and are transcribed off opposite DNA strands in five other chloroplast genomes. These differences are probably the consequence of an approximately 50 kb inversion which distinguishes the pea and mung bean genomes from those of most other angiosperms (1).

Biological Evolution↗
PubMed1999

Glutathione and homoglutathione synthesis in legume root nodules.

High-performance liquid chromatography (HPLC) with fluorescence detection was used to study thiol metabolism in legume nodules. Glutathione (GSH) was the major non-protein thiol in all indeterminate nodules examined, as well as in the determinate nodules of cowpea (Vigna unguiculata), whereas homoglutathione (hGSH) predominated in soybean (Glycine max), bean (Phaseolus vulgaris), and mungbean (Vigna radiata) nodules. All nodules had greater thiol concentrations than the leaves and roots of the same plants because of active thiol synthesis in nodule tissue. The correlation between thiol tripeptides and the activities of glutathione synthetase (GSHS) and homoglutathione synthetase (hGSHS) in the nodules of eight legumes, and the contrasting thiol contents and activities in alfalfa (Medicago sativa) leaves (98% hGSH, 100% hGSHS) and nodules (72% GSH, 80% GSHS) indicated that the distribution of GSH and hGSH is determined by specific synthetases. Thiol contents and synthesis decreased with both natural and induced nodule senescence, and were also reduced in the senescent zone of indeterminate nodules. Thiols and GSHS were especially abundant in the meristematic and infected zones of pea (Pisum sativum) nodules. Thiols and gamma-glutamylcysteinyl synthetase were also more abundant in the infected zone of bean nodules, but hGSHS was predominant in the cortex. Isolation of full-length cDNA sequences coding for gamma-glutamylcysteinyl synthetase from legume nodules revealed that they are highly homologous to those from other higher plants.

Amino Acid Sequence↗
PubMed1969

Relative activities of NAD- and NADP- isocritric dehydrogenases in bean mitochondria modified by glycerol or NADP.

Mitochondria from cotyledons of Vigna sesquipedalis (L.) Fruwirth (starchy seed) showed no NAD-isocitric dehydrogenase (NAD-IDH) activity by the methods which have been known to be useful for the detection of NAD-IDH in mitochondria of plants including castor bean and alaska pea. When the Vigna cotyledon mitochondria were treated with glycerol, NAD-IDH activity appeared and NADP-isocitric dehydrogenase (NADP-IDH) activity was inhibited. The inhibition of mitochondrial NADP-IDH by glycerol was overcome by the addition of excess NADP. On the other hand, NADP-IDH activity in the soluble fraction of cell components was only slightly inhibited by glycerol and no NAD-IDH activity was elicited. It was postulated that NADP-IDH in mitochondria is converted to NAD-IDH by glycerol and back to NADP-IDH with NADP by the alteration in the spatial configuration of the enzyme. However, there could be 2 proteins as the other possibility. The NADP-IDH in the soluble fraction which is not subject to such alteration is different from the mitochondrial NADP-IDH.

Glycerol↗
PubMed1979

Legume Lectins: I. Immunological Cross-Reactions between the Enzymic Lectin from Mung Beans and other Well Characterized Legume Lectins.

A number of well characterized legume lectins including the enzymic lectin from Vigna radiata were examined for immunological relatedness. The immunological cross-reactions observed indicate that most of the legume lectins, including Vigna lectin, are evolutionarily closely related proteins. The possibility that these proteins are homologs with enzymic functions is discussed.

Journal Article↗
PubMed1979

Enzyme Profiles in Seedling Development and the Effect of Itaconate, an Isocitrate Lyase-directed Reagent.

Changes in levels of isocitrate lyase, malate synthase, and catalase have been investigated during germination of flax (Linum usitatissimum L.) in the presence and absence of itaconate. Germination was accompanied by a rapid increase in these enzymes during the first 3 days. The presence of 38 millimolar itaconate inhibited the incidence of seed germination and the growth of embryo axes as well as the appearance of isocitrate lyase but did not alter the levels of malate synthase, catalase, or NADP(+)-isocitrate dehydrogenase. The specific activity for the latter enzyme was constant throughout germination. Oxalate or succinate, each at 38 millimolar, had no effect upon germination of flax seeds. Itaconate did not inhibit the activities of malate synthase, catalase, or NADP(+)-isocitrate dehydrogenase in vitro but was a potent noncompetitive inhibitor of isocitrate lyase (K(i):17 micromolar at 30 C, pH 7.6). Itaconate (at 38 millimolar) did not alter the appearance of malate synthase but reduced the incidence of germination, onset of germination, and growth of the embryo axis as well as the specific activity of isocitrate lyase in seedlings of Zea mays, Vigna glabra, Glycine hispida, Vigna sinensis, Trigonella foenumgraecum, Lens culinaris, and Medicago sativa. The incidence and onset of germination of wheat seeds were unaltered by the same concentration of itaconate but seedlings did not contain isocitrate lyase or malate synthase. The data suggest that itaconate may be isocitrate lyase-directed in inhibiting the germination of fatty seeds.

Journal Article↗