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PubMed1986

Cationic form of beta-galactosidase in the germinating seeds of Vigna sinensis (Linn) Savi.

The cationic form of beta-galactosidase (EC 3.2.1.23) from the germinating seeds of Vigna sinensis has been separated from its other isoforms by DEAE-cellulose (DE-52) column chromatography and further purified by gel filtration and affinity chromatography. Polyacrylamide gel electrophoresis of the purified enzyme imparted a single protein band. The molecular mass of the enzyme as determined by Sephadex G-150 gel filtration is 58,800 Da. The optimum temperature and the optimum pH are 60 degrees C and 4.5, respectively. Most of the metal ions tested were inhibitory to the enzyme activity. The enzyme has Km for p-nitrophenyl beta-D-galactoside and o-nitrophenyl beta-D-galactoside of 0.56 and 2.0 mM, respectively. The Ki values of galactose and lactose are 2.4 and 70.0 mM, respectively. The energy of activation of PNPG for the enzyme is 10.3 kcal/mol.

Galactosidases↗
PubMed1994

Purification and characterization of a soluble lipolytic acylhydrolase from Cowpea (Vigna unguiculata L.) leaves.

With the use of [14C]monogalactosyl diacylglycerol as substrate for enzymatic test, a lipolytic acylhydrolase (EC 3.1.1.26) was purified 263-fold with a yield of 2.0% from soluble leaf extract of Vigna unguiculata L. cv. EPACE-1. The procedure involved ammonium sulfate precipitation, Q-Sepharose Fast Flow chromatography, gel filtration on Sephacryl 300 HR and chromatofocusing on Mono-P, followed by a semi-preparative electrophoresis on polyacrylamide gel. The purified enzyme had a molecular mass of about 80 kDa, as determined by gel filtration. On SDS-PAGE, it showed a single band corresponding to a molecular mass of 40 kDa. The isoelectric point of the enzyme was estimated to be 5.0-5.1 by isoelectric focusing and chromatofocusing. The Km value was 0.119 mM for monogalactosyl-diacylglycerol. The hydrolytic activity of the enzyme on different substrates was determined: the relative rates were digalactosyl-diacylglycerol > monogalactosyl-diacylglycerol > phosphatidylcholine > phosphatidylglycerol. For all substrates, the products of hydrolysis were free fatty acids. Triacylglycerols were not hydrolysed. The enzyme was activated by calcium but was not calcium-dependent. Experiments concerning the enzyme stability as affected by temperature and pH demonstrated that it was quite stable.

Carboxylic Ester Hydrolases↗
PubMed1986

Construction of a genomic library from germinating seedlings of mung bean (Vigna radiata). Evidence for the presence of a class of repetitive sequences and selection of beta-tubulin specific recombinant phage.

We have constructed a genomic library of Vigna radiata from its total DNA in the phage charon 4A. Recombinant phages were used to hybridize with a family of mung bean repetitive DNA. At least 30% of recombinant phages showed positive hybridization with a specific type of sequences. These phages were also used to screen for beta-tubulin-specific recombinants and a recombinant phage with beta-tubulin gene was identified.

Autoradiography↗
PubMed1996

Purification of acid phosphatase I from germinating seeds of Vigna sinensis.

Acid phosphatase I (AP-I) is the major isoform of Vigna acid phosphatase. It is constitutively expressed in seed cotyledons during germination. AP-I was separated from other isoforms and purified to homogeneity by three simple purification steps; (NH4)2SO4 precipitation, and phosphocellulose and DEAE-cellulose column chromatography. The activity of AP-I was not affected by 1 mM Mg2+, Mn2+, Ca2+, Co2+ or Pb2+, but severely inhibited by 1 mM Cu2+, Fe3+, Hg2+, Mo6+ or Zn2+. AP-I has both phosphatase and pyrophosphatase activities, and is highly stable even at 50 degrees.

Acid Phosphatase↗
PubMed1993

Pyrophosphate: fructose-6-phosphate 1-phosphotransferase and biosynthetic capacity during differentiation of hypocotyls of Vigna seedlings.

The relationship between the activity of pyrophosphate:fructose-6-phosphate 1-phosphotransferase (PFP) and the capacity for biosynthesis of macromolecules was examined in segments from different parts of hypocotyls of etiolated seedlings of Vigna mungo and V. radiata. The relative ratio of the maximum activity of PFP to that of ATP-dependent phosphofructokinase (PFK) (PFP/PFK ratio) was high in young tissues and decreased with differentiation and ageing of the tissues. The highest level of fructose-2,6-bisphosphate was observed in the youngest part of hypocotyls of V. mungo. The level was markedly decreased with ageing of tissues. The levels of PPi and ATP were also higher in younger parts than in older parts of the hypocotyls, but the ratio of the level of PPi to that of ATP was almost constant in all parts of the hypocotyl. A good correlation was found between the PFP/PFK ratio and the biosynthetic capacity, as estimated from the rate of incorporation of [U-14C]sucrose into ethanol-insoluble macromolecules.

Aging↗
PubMed1988

Isolation, characterization and sequencing of a novel repetitive DNA from the mung bean Vigna radiata.

A family of highly reiterated, small (approx. 300 bp) sequences has been identified in DNA of the mung bean Vigna radiata. The members are extensively interspersed throughout the chromosomes with some clustering. They also occur extrachromosomally. There is no tissue-specificity to the repeat family but it is highly species-specific. The repetitive DNA hybridizes to total RNA as well as to polyadenylated RNA isolated from germinated mung beans. It has analogy with the human AluI family in the mode of isolation, size, genomic distribution, copy number and transcribability though they do not share any sequence homology. A repetitive DNA clone was selected from a shotgun genomic library of mung bean DNA in pBR322. The average copy number of the cloned repeat is estimated to be 8 x 10(4) per haploid genome, and thus constitutes approx. 5% of the total mung bean genome. The genomic organization and transcription of the cloned repeat is reported. Sequencing of the cloned repetitive DNA reveals the presence of the number of direct and inverted repeats and some short palindromic sequences.

Base Sequence↗
PubMed1988

Sequence organization and putative regulatory elements in the 5S rRNA genes of two higher plants (Vigna radiata and Matthiola incana).

The tandemly arranged and clustered highly repeated 5S rRNA genes are investigated for two plants belonging to different higher plant families: Matthiola incana (Brassicaceae, Dilleniidae, Rosidae; 3600 5S rRNA genes/n) shows a homogeneous repeat size of 510 bp, whereas Vigna radiata (mung bean, former Phaseolus aureus, Fabaceae, Rosidae; approx. 4300 5S rRNA genes) has a repeat size of 215 bp. The mung-bean 5S rRNA coding region starts 5' with AGG and ends with CCT; Matthiola starts with GGG and ends with CCC. Striking is the strict occurrence of a 'TATA' box starting at nucleotide-28 similar to Neurospora crassa 5S rRNA genes. The 3' end is followed by CTTTT or GTTT stretches present in different numbers in the non-transcribed spacer suggesting a function in termination.

Base Sequence↗
PubMed2004

Study of salt permeation process into Vigna angularis using Ag/AgCl electrodes.

In this work we report a novel method that can monitor in vivo the salt permeation process into a plant Vigna angularis using two Ag/AgCl electrodes. The method is based on the electromotive force (emf) measurement using two Ag/AgCl electrodes, one inserted into the V. angularis pith cavity and the other placed into saline ([NaCl]=1 M) surrounding the roots. Temporary change of emf ranging from 0 to ca. 0.115 V was measured in vivo using the system, and the possible physiological phenomenon during the period was discussed.

Electrochemistry↗
PubMed2005

Effect of seaweed extracts on the growth and biochemical constituents of Vigna sinensis.

The effect of seaweed liquid fertilizers (SLF) of Sargassum wightii and Caulerpa chemnitzia on growth and biochemical constituents of Vigna sinensis was studied. The seeds soaked with aqueous extract of seaweeds performed better when compared to the water soaked controls. Hundred per cent germination was recorded both in aqueous extract soaked and water soaked treatments. The low concentration (20%) of aqueous extracts of S. wightii and C. chemnitzia promoted the seedling growth including the parameters of shoot length (15.87, 14.13 cm/seedling), root length (6.42, 5.38 cm/seedling), fresh weight (4.017, 4.012 g/seedling) and dry weight (0.878, 0.865 g/seedling), chlorophyll (1.599, 1.491 mg g-1 fr. wt.), carotenoids (0.899, 0.875 mg g-1 fr. wt.), protein content of shoot (3.956, 3.474 mg g-1 fr. wt.) and root (2.926, 2.890 mg g-1 fr. wt.), amino acid content of shoot (1.447, 1.429 mg g-1 fr. wt.) and root (0.698, 0.680 mg g-1 fr. wt.), reducing sugar content of shoot (6.426, 6.233 mg g-1 fr. wt.) and root (5.118, 5.103 mg g-1 fr. wt.), total sugar content of shoot (11.846, 11.350 mg g-1 fr. wt.) and root (10.368, 10.102 mg g-1 fr. wt.), alpha-amylase (1.927, 1.819 microg min-1 mg-1 protein) and beta-amylase (1.730, 1.617 microg min-1 mg-1 protein) activities in V. sinensis. Among the two seaweeds tested, S. wightii exhibited better responses.

Amino Acids↗
PubMed2006

Isolation and structural analysis of ajugose from Vigna mungo L.

The hexasaccharide ajugose, alpha-D-galactopyranosyl-(1-->6)-alpha-D-galactopyranosyl-(1-->6)-O-alpha-D-galactopyranosyl-(1-->6)-alpha-D-galactopyranosyl-(1-->6)-alpha-D-glucopyranosyl-(1<-->2)-beta-D-fructofuranoside, generally uncommon in legumes, was detected in the seeds of Vigna mungo L. by TLC and paper chromatography. Ajugose was then isolated by silica gel chromatography and its structure was established by acid and enzymatic hydrolysis, fast atom bombardment mass spectrometry and both one- and two-dimensional 1H and 13C NMR techniques.

Carbohydrate Sequence↗
PubMed2005

Role of ethylene diurea (EDU) in assessing impact of ozone on Vigna radiata L. plants in a suburban area of Allahabad (India).

A field study was conducted to evaluate the suitability of ethylene diurea (N-[2-(2-oxo-1-imidazolidinyl)ethyl]-N'-phenylurea; EDU) in assessing the impact of O3 on mung bean plants (Vigna radiata L. var. Malviya Jyoti) grown in suburban area of Allahabad city situated in a dry tropical region of India. EDU is a synthetic chemical having anti-ozonant property. Mean monthly O3 concentration varied between 64 and 69 microg m(-3) during the experimental period. In comparison to EDU-treated plants, non-EDU-treated plants showed significant reductions in plant growth and yield under ambient conditions. Significant favourable effects of EDU-application were observed with respect to photosynthetic pigments, soluble protein, ascorbic acid and phenol contents. EDU-treated plants maintained higher levels of pigments, protein and ascorbic acid in foliage as compared to non-EDU-treated ones. The study clearly demonstrated that EDU alleviates the unfavourable effects of O3 on mung bean plants, and therefore can be used as a tool to assess the growth and yield losses in areas having higher O3 concentrations.

Agriculture↗
PubMed2006

Toxic effects of low concentrations of Cu on nodulation of cowpea (Vigna unguiculata).

Although Cu is phytotoxic at Cu(2+) activities as low as 1-2 microM, the effect of Cu(2+) on the nodulation of legumes has received little attention. The effect of Cu(2+) on nodulation of cowpea (Vigna unguiculata (L.) Walp. cv. Caloona) was examined in a dilute solution culture system utilising a cation exchange resin to buffer solution Cu(2+). The nodulation process was more sensitive to increasing Cu(2+) activities than both shoot and root growth; whilst a Cu(2+) activity of 1.0 microM corresponded to a 10% reduction in the relative yield of the shoots and roots, a Cu(2+) activity of 0.2 microM corresponded to a 10% reduction in nodulation. This reduction in nodulation with increasing Cu(2+) activity was associated with an inhibition of root hair formation in treatments containing > or =0.77 microM Cu(2+), rather than to a reduction in the size of the Rhizobium population.

Copper↗
PubMed2004

A multicystatin is induced by drought-stress in cowpea (Vigna unguiculata (L.) Walp.) leaves.

Cystatins are protein inhibitors of cystein proteinases belonging to the papain family. In cowpea, cystatin-like polypeptides and a cDNA have been identified from seeds and metabolic functions have been attributed to them. This paper describes VuC1, a new cystatin cDNA isolated from cowpea leaves (Vigna unguiculata (L.) Walp.). Sequence analysis revealed a multicystatin structure with two cystatin-like domains. The recombinant VUC1 protein (rVUC1) was expressed in an heterologous expression system and purified to apparent homogeneity. It appeared to be an efficient inhibitor of papain activity on a chromogenic substrate. Polyclonal antibodies against rVUC1 were obtained. Involvement of the VuC1 cDNA in the cellular response to various abiotic stresses (progressive drought-stress, dessication and application of exogenous abscissic acid) was studied, using Northern blot and Western blot analysis, in the leaf tissues of cowpea plants corresponding to two cultivars with different capacity to tolerate drought-stress. Surprisingly, these abiotic stresses induced accumulation of two VuC1-like messages both translated into VUC1-like polypeptides. Difference in the transcript accumulation patterns was observed between the two cultivars and related to their respective tolerance level. Presence of multiple cystatin-like polypeptides and their possible involvement in the control of leaf protein degradation by cysteine proteinases is discussed.

Amino Acid Sequence↗
PubMed2005

Protective effect of polyphenol-containing azuki bean (Vigna angularis) seed coats on the renal cortex in streptozotocin-induced diabetic rats.

This study was undertaken to investigate the effect of azuki bean (Vigna angularis) seed coats (ABSC), which contain polyphenols, on the infiltration of macrophages and the progression of diabetic nephropathy in streptozotocin (STZ)-induced diabetic rats. The diabetic rats were divided into three groups with 0% (commercial diet), 0.1% and 1.0% ABSC diets. The vehicle-injected controls were given a commercial diet. At 10 weeks, the macrophage kinetics, the degree of fibrosis in glomeruli and mRNA expression for monocyte chemoattractant protein-1 (MCP-1) were examined. There was no difference in plasma glucose levels between diabetic rats treated with and without ABSC. The plasma levels of malondialdehyde (MDA) in the ABSC-treated diabetic rats were significantly lower than those in the untreated diabetic rats. Histopathologically, the percentage of the fibrotic areas stained by Sirius red stain in the glomeruli in the ABSC-treated diabetic rats was lower than in the untreated diabetic rats. ED1-positive macrophages in the glomeruli and tubulointerstitium in the untreated diabetic rats showed a significant increase in number compared with the controls. In contrast, the number of macrophages in the ABSC-treated diabetic rats was smaller than that in untreated diabetic rats. MCP-1 mRNA expression, estimated by real-time quantitative RT-PCR, was increased 2.5-fold in the untreated diabetic rat kidney, while a lower level was observed in the ABSC-treated diabetic rats. In conclusion, our results suggest that ABSC treatments suppress the increased number of infiltrating macrophages and MCP-1 mRNA expression, and attenuated the glomerular expansion in STZ-induced rat diabetic nephropathy.

Animals↗
PubMed2004

Isolation and characterization of an aspartic proteinase gene from cowpea (Vigna unguiculata L. Walp.) .

A cowpea (Vigna unguiculata cv. EPACE-1) aspartic proteinase (AP) gene was isolated by genomic Library screening. Sequence analysis shows that this AP gene follows the same pattern of intron/exon number and organization as the other isolated plant AP genes, which are distinct from other solved AP genes. Northern blot analysis revealed that cowpea AP accumulates in leaves and stems but not in roots, indicating tissue-specific expression. An increased accumulation of transcripts during senescence suggests enzyme involvement in this process.

Amino Acid Sequence↗
PubMed2006

Alternative oxidase regulation in roots of Vigna unguiculata cultivars differing in drought/salt tolerance.

The alternative oxidase (Aox) was studied at different levels (transcript, protein and capacity) in response to an osmotic shock applied to roots of cowpea (Vigna unguiculata). Two cultivars of V. unguiculata were used, Vita 3 and Vita 5, tolerant and sensitive to drought/saline stress respectively. The seedlings (17-day-old) were grown in hydroponic conditions and submitted to NaCl (100 and 200 mM) or 200.67 g L(-1) PEG 6000 (iso-osmotic condition to 100 mM NaCl). The VuAox1 and VuAox2a mRNA were not detected in either cultivar under all tested conditions while the VuAox2b gene was differently expressed. In the tolerant cultivar (Vita 3), the expression of VuAox2b gene was stimulated by an osmotic stress induced by PEG which was associated with a higher amount and capacity of the Aox protein. In the same cultivar, this gene was under-expressed in salt stress conditions with poor effect on the protein level. In the sensitive cultivar (Vita 5), the transcript level of the VuAox2b was unchanged in response to PEG treatment, even though the protein and the capacity tended to increase. Upon salt stress, the VuAox2b gene was over-expressed. At 100mM NaCl, this VuAox2b gene over-expression led to a higher amount and capacity of Aox. This effect was reduced at 200 mM NaCl. Overall, these results suggest complex mechanisms (transcriptional, translational and post-translational) for Aox regulation in response to osmotic stress.

Fabaceae↗
PubMed2005

Protective effect of dietary azuki bean (Vigna angularis) seed coats against renal interstitial fibrosis of rats induced by cisplatin.

OBJECTIVE: We investigated the effects of azuki bean (Vigna angularis) seed coats (ABSCs), which mainly contain proanthocyanidins and dietary fibers, on the infiltration of macrophages and the progression of renal interstitial fibrosis induced by cisplatin (CDDP). METHODS: Male rats were divided into two groups: controls received intraperitoneal injections of saline and the other rats were injected intraperitoneally with 3 mg of CDDP per kilogram once a week for 5 wk. The CDDP-injected animals received one of four diets: 1) control diet (commercial diet only), 2) 0.5% red ABSC (RABSC) diet, 3) 2.0% RABSC diet, and 4) 2.0% white ABSC (WABSC) diet. The saline-injected animals were given the commercial diet. Five weeks after the final CDDP injection, macrophage kinetics and interstitial fibrotic areas were examined. RESULTS: The content of polyphenols in the RABSC (76.3 g/kg of plant material) was higher than that in the WABSC (18.1 g/kg). Proanthocyanidins were detected in the RABSC (20.4 g/kg) but not in the WABSC. Histologically, the fibrotic areas consisting of fibroblastic cells and mononuclear cells developed around the dilated or atrophic tubules in the corticomedullary junction in CDDP-treated rat kidney, whereas the extent and magnitude of damage were less in the WABSC- and RABSC-treated rats. In immunohistochemical analysis, ED1-positive macrophages in CDDP-treated rats showed a significant increase in number compared with the control. The number of macrophages in CDDP plus WABSC or RABSC groups was significantly smaller than that in CDDP-treated rats. In addition, the number of macrophages in the RABSC group was significantly smaller than in the WABSC group, indicating that ABSC, especially RABSC, prevented macrophages from infiltrating into areas of interstitial fibrosis. CONCLUSIONS: These results suggest that ABSC, especially RABSC, suppress the increase of infiltrating macrophages in the damaged kidney and may lead to amelioration of interstitial fibrosis. Based on the composition of ABSC, molecules such as proanthocyanidins and/or dietary fibers may be associated with the amelioration of renal damage.

Animals↗
PubMed2005

A beta-(1-->3)-arabinopyranosyltransferase that transfers a single arabinopyranose onto arabino-oligosaccharides in mung bean (Vigna radiate) hypocotyls.

Arabinopyranosyltransferase (ArapT) activity that results in the transfer of a single arabinopyranose (Arap) residue from UDP-beta-L-arabinopyranose (UDP-Arap) to exogenous (1-->5)-linked alpha-L-arabino-oligosaccharides labeled with 2-aminobenzamide (2-AB) at their reducing ends was identified in a particulate preparation obtained from 3-day-old mung bean (Vigna radiate L. Wilezek) hypocotyls. The transferred Ara residue was shown to be beta-(1-->3)-linked to O-3 of the non-reducing terminal Araf residues of the oligosaccharide using nuclear magnetic resonance spectroscopy together with glycosyl composition and glycosyl linkage composition analyses. The 2AB-labeled arabino-octasaccharide was the most effective acceptor substrate analyzed, although arabino-oligosaccharides with a degree of polymerization between 4 and 7 were also acceptor substrates. Maximum ArapT activity was obtained at pH 6.5-7.0, and 20 degrees C in the presence of 25 mM Mn(2+) and 0.5% Triton X-100.

Arabinose↗