Mungbean [Vigna radiata (L.) Wilczek] globulins: purification and characterization

Vicilin type (8S) and basic 7S globulins and legumin type (11S) globulins were isolated from mungbean [Vigna radiata (L.) Wilczek]. The native molecular weights of the different globulin types were 360000 for legumin, 200000 for vicilin, and 135000 for basic 7S. Some of the 8S globulin apparently complexed and coeluted with the 11S on gel filtration. On SDS-PAGE, 11S was composed of two bands of 40000 and 24000, 8S was composed of 60000, 48000, 32000, and 26000 bands, and basic 7S was composed of 28000 and 16000 bands. The percent composition of total globulins was estimated to be as follow: 8S, 89%; basic 7S, 3.4%; and 11S, 7.6%. The basic 7S and 11S but not the 8S globulins were found to have disulfide bonds. The presence of carbohydrates by conjugated peroxidase reaction was observed in all bands of 8S, the acidic polypeptide of basic 7S, and its complex but not in 11S. The 28000 basic 7S band and its 42000 complex and the first three major bands of 8S cross-reacted with antibodies to all types of soybean conglycinin subunits (alpha, alpha', and beta), whereas the fourth band cross-reacted only with the anti-beta subunit. None of the mungbean globulins cross-reacted with anti-soybean glycinin. Basic 7S was found to be easily extracted with 0.15 M NaCl, 11S was extracted with 0.35 M NaCl,and 8S was extracted over a wide range of NaCl concentrations. The N-terminal sequences of the different subunits/fragments of the globulins were determined and found to have strong homology with storage proteins of other legumes and crops.     

8S globulin of mungbean [Vigna radiata (L.) Wilczek]: cloning and characterization of its cDNA isoforms, expression in Escherichia coli, purification, and crystallization of the major recombinant 8S isoform

Three isoforms of the cDNA of the major 8S globulin of mungbean, 8Salpha, 8Salpha', and 8Sbeta, were isolated, cloned, and characterized. The cDNA sequences of 8Salpha, 8Salpha', and 8Sbeta had open reading frames of 1362, 1359 or 1362, and 1359 bp, respectively, which code for 454, 453 or 454, and 453 amino acids corresponding to molecular weights of 51 973, 51 627 or 51 758, and 51 779, respectively. Homology in terms of cDNA and amino acid sequences was 91-92% between 8Salpha and 8Salpha', 87% between 8Salpha and 8Sbeta, and 86-88% between 8Salpha' and 8Sbeta. The signal peptide was found to be 1-25, 1-24 or 25, and 1-23 for 8Salpha, 8Salpha', and 8Sbeta, respectively, using the signalP website (Nielsen, H.; Engelbrecht, J.; Brunak, S.; von Heijne, G. Protein Eng. 1997, 10, 1-6). The propeptide was determined to be IVHREN. A single site for glycosylation (N-X-S/T) was observed about 90 amino acids from the C terminus. Homology between mungbean 8S isoforms and other 7-8S proteins ranged from 45 to 68% within members of the legume family and 29 to 34% for crops of different species. The major isoform 8Salpha was expressed in Escherichia coli and purified by successive ammonium sulfate fractionation, hydrophobic interaction, and Mono Q column chromatography. The recombinant 8Salpha, but not the native form, was successfully crystallized producing rhombohedral crystals.     

Salt stress and mungbean [Vigna radiata (L.) Wilczek]: effects,physiological perspective and management practices for alleviating salinity

Mungbean is an important food grain legume with high economic status. It has an excellent source of dietary protein and nutritional health benefits, particularly for the vegetarians. It increases soil fertility and also plays an imperative role in major cropping systems due to its short life span. Production of mungbean is still decreasing due to its susceptibility towards various environmental stress factors. Salt stress is one of the most prevailing abiotic stress imposing threats for agriculture food crops along with increasing world population and limited natural resources. Fewer efforts have been made to develop an improved variety of mungbean. The present review summarizes the adverse effects of salt stress and mungbean response at the physiological and molecular level. It covers recent studies on introgression of useful traits in mungbean for its better adaptability and survival under stressed conditions. Modern biotechnological approaches and traditional breeding methods may assist the development of salt-tolerant cultivars of mungbean for salinity-affected area in arid and semi-arid regions. Researchers involved in this area should keep this goal on priority for sustainable mungbean production. Availability of protein-rich food may help to reduce the problem of malnutrition in poor families and national food security issue for a continuous rising population.     

The identification of starch phosphorylase in the developing mungbean (Vigna radiata L.)

Starch phosphorylase (SP) in immature mungbean (Vigna radiata L. cv KPS1) seed soluble extract was detected by in situ activity staining and identified by MALDI-TOF mass analysis. After in situ SP assay on native-PAGE, a major starch-enzyme complex was located on the gel zymogram in a dose-dependent manner. This complex depicted two major SP-activity related proteins, 105 kDa and 55 kDa, by SDS-PAGE. The mass and predicted sequence of the tryptic fragments of the isolated 105 kDa protein, analyzed by MALDI-TOF spectroscopy and bioinformatic analysis, confirmed it to be mungbean SP as a result of high similarity to the L-SP of known plant. Polyclonal antibodies raised from the 55 kDa recognized both the 105 kDa and the 55 kDa proteins on the Western blot and neutralized partial SP activity, indicating that the two proteins were immunologically related. The 55 kDa protein possess high similarity to the N-terminal half of the 105 kDa SP was further confirmed. The SP activity and the activity stained protein density in mungbean soluble extract decreased as the seed size increased during early seed growth. These data indicate that mungbean 105 kDa SP and SP activity-related 55 kDa were identified in the developing mungbean.     

Cobalt reduces nitrate inhibition of nodulation in mungbean (Vigna radiata)

 A cobalt-mediated decrease in ethylene production reduced the inhibition of nodulation by nitrate in Vigna radiata (mungbean). Nitrate increased the ethylene production in 5-day-old seedlings, while it caused a reduction in the nodulation status (nodule number and nodule weight) and nodule efficiency (acetylene reduction activity) in mungbean plants. The application of cobalt chloride inhibited nitrate-affected ethylene production and also decreased the inhibitory effect of nitrate on nodulation. The effect of cobalt was most marked on nodule number. Received: 6 August 1999     

Effect of vesicular-arbuscular mycorrhizae and zinc application on dry matter and zinc uptake of greengram (Vigna radiata L. Wilczek)

Summary In a greenhouse study we examined the effects of vesicular-arbuscular mycorrhizae (VAM) inoculation, using Glomus macrocarpum and of Zn application on dry matter production and Zn uptake by greengram in two mollisols. The VAM inoculation significantly increased the dry weight of different plant parts and the Zn uptake in both soils. Inoculated plants showed a greater response to the application of Zn at 2.5 and 5.0 mg kg^-1 soil in a Zn deficient clay loam soil. The inoculated plants also absorbed — more water than the uninoculated plants. Mass flow and diffusion were the principal processes by which Zn reached the plant roots; mass flow was particularly important in the absence of VAM in a sandy soil fertilized with higher Zn doses (5 and 10 mg kg^-1 soil). The greater supply of Zn to inoculated roots was attributed to an apparent diffusion process rather than to mass flow of Zn.     

Detection of proteins related to starch synthase activity in the developing mungbean (Vigna radiata L.)

Proteins associated with starch synthase (SS) activities were identified in immature mungbeans (Vigna radiata L. cv KPS1). Seed soluble extract was separated by native-PAGE and subjected to in situ activity staining. The gel zymogram located starch-enzyme complex bands. The soluble extract was also partitioned by preparative-IEF and screened for SS activity using radioactive assay. IEF fractions eluted within pH 4-6 revealed enriched SS activity of 145-fold. Parallel comparison of the protein profiles among the activity stained enzyme complex and the active isoelectric focused fractions on SDS-PAGE depicted three SS-activity-related proteins with molecular size of 32, 53, and 85 kDa. The 85 kDa protein, however, was identified to be methionine synthase by MALDI-TOF analysis and should be a protein physically associated with the active SS. Polyclonal antibodies raised from eluted native enzyme complex neutralized up to 90% activity and antigenically recognize the other 53 and 32 kDa proteins on Western blot. Antibodies raised from the two individual denatured proteins were able to neutralize SS activities near 60% separately, indicating that the 53 kDa and 32 proteins associated with SS activity are potentially involved in starch biosynthesis during mungbean seed development.     

Residual effects of nitrogen sources,sulfur and boron levels on mungbean (Vigna radiata) in a sunflower (Helianthus annuus)–mungbean system

Field experiments were conducted during spring–rainy (kharif) seasons of 2005 and 2006 on a sunflower–mungbean cropping system at the research farm of the Division of Agronomy, Indian Agricultural Research Institute (IARI), New Delhi, India. The objectives of this study were to investigate the residual effect of nitrogen sources, sulfur and boron levels applied to sunflower on productivity, nutrient concentrations and their uptake by the succeeding mungbean crop in a sunflower–mungbean cropping system. The experiment with 19 treatments was laid out in factorial randomized block design for both sunflower and mungbean. The residual effects of nutrients applied to sunflower were significant on the succeeding mungbean crop in terms of biometric parameters, yield attributing characters, seed yield and soil nutrient status. The highest mungbean seed yield (961.2 kg ha^?1) was produced with 50 kg ha^?1 sulfur application to the preceding sunflower crop, which was significantly (p < 0.05) higher than with 0 and 25 kg sulfur ha^?1. The concentrations and uptake of nitrogen, sulfur and boron were also greater in the succeeding mungbean crop due to the residual effects of nutrients applied to the preceding sunflower crop. The soil nutrient status before and after mungbean indicated that the available nitrogen and sulfur were higher due to application to the preceding crop, while available boron after mungbean was even higher than after sunflower due to its slow release and static nature in the soil.     

Irrigation with brackish water modifies the boron requirement of mungbean (Vigna radiata L.) on typic calciargid

The boron (B) sufficiency range for plant growth is narrow and its management is problematic under brackish irrigation water. This study was conducted to evaluate the B requirement of mungbean at different sodium adsorption ratios of irrigation waters (SAR_iw) [control, 8 and 16 (mmol_c L^?1)^1/2]. The boron adsorption characteristics of a loamy soil were first determined in the laboratory by equilibrating 2.5 g soil with 0.01 M CaCl₂ solution containing different B levels. Boron rates for a pot study were computed against different soil solution levels by fitting sorption data in a modified Freundlich model [x/m = K _f (EBC)^1/n ]. The maximum increase in shoot dry matter was 11.9% when B was applied at 1.29 mg kg^?1 soil at control SAR_iw. Visual leaf B toxicity symptoms appeared at higher B rates and became severe at higher SAR_iw. By contrast to Ca, shoot concentrations of B and Na increased significantly with B application and SAR_iw. For optimum shoot growth, internal and external B requirements were 25 mg B kg^?1 shoot dry matter and 0.39 mg B L^?1 soil solution, respectively, at control SAR_iw. At higher SAR_iw, a lower concentration of B in plant shoots and soil solution had an inhibitory effect on plant growth.     

Plant analysis diagnostic indices for boron nutrition of mungbean (Vigna radiata L.) cultivars grown in a rainfed calcareous soil

Boron requirement of mungbean was assessed from yield response curves based on field experiment during 2010-11. Four cultivars, i.e., NM-11, NM-2006, Chakwal Mung-2006 and AZRI Mung-2006 were grown in a boron (B)-deficient calcareous soil by applying B at 0, 0.5, 1.0, 2.0, and 4.0 kg ha^?I. Boron application significantly increased grain yield and B uptake by all the mungbean cultivars over control. Maximum grain yield increases were: cv. NM-11, 20%; cv. NM-2006, 24%; cv. Chakwal Mung-2006, 26%; and cv. AZRI Mung-2006, 28%. The cultivars varied in B efficiency and cv. NM-11 was most B efficient (83%) while cv. AZRI Mung-2006 was the least B efficient (78%). Fertilizer B requirement (kg ha^?1) for 95% of the maximum grain yield was: 0.60 for cv. AZRI Mung-2006, 0.70 for cv. Chakwal Mung-2006, 0.75 for cv. NM-2006, and 0.85 for cv. NM-11. Plant tissue critical B concentration in leaves was higher than in seeds.     

Effects of different phosphatase-producing fungi on growth and nutrition of mung beans [Vigna radiata (L.) Wilczek] in an arid soil

Summary The efficiency of different phosphatase-producing fungi in arid soils was studied in pot experiments in order to evaluate the effect of these organisms on the enhancement of rhizosphere enzyme activities, the uptake of different nutrients, dry matter production, and grain yields of the mung bean. A significant increase in phosphatase (acid and alkaline), nitrogenase, and dehydrogenase activities was observed upon inoculation. Inoculation with phosphatase-producing fungi significantly increased dry matter production and grain yields compared with uninoculated controls. In general, there was a significant improvement in the uptake of N, P, K, Ca, Mg, Fe, and Zn while no effect on Na, Mn, and Cu was recorded. A significant positive correlation was observed between the activities of various enzymes in the rhizosphere on the one hand and N and P uptake, dry matter production, and grain yields of the mung bean on the other. Aspergillus rugulosus was the best of the phosphatase-producing fungi that were tested, followed by A. terreus. The experiment confirmed that phosphatase-producing fungi have a significant effect on growth and nutrient uptake in the mung bean and indicated that these fungi are particularly important in arid soils.     

Response of maize (Zea mays L.) and mungbean (Vigna radiata L.) to tillage in relation to water table depth in tropical lowland rice soils

The effects of zero, minimum and conventional tillage on soil physical properties and on the growth and yield of mungbean (Vigna radiata L.) grown after lowland rice (Oryza sativa L.) were studied in field experiments conducted during the 1984 and 1985 dry seasons (DS) at two Philippine sites (clay loam, Vertic Tropaquept, with shallow water table and sandy loam, Aeric Tropaquept, with deep water table). Effects on maize (Zea mays L.) were studied only in 1984 on clay loam soil.All parameter measurements were not significantly different with minimum and conventional tillage. Tillage, averaged over minimum and conventional and in both seasons, significantly lowered bulk density (10%) and increased aeration porosity (120%) of the 0–0.10 m clay loam soil layer. In sandy loam soil in 1985, it decreased bulk density by 7% and increased aeration porosity by 61%. Tillage only slightly affected the matric suction, strength and temperature of both soils.Maize seedling emergence was 15% higher with zero tillage than with minimum and conventional tillage. Tillage, however, did not affect mungbean emergence. It significantly increased maize plant height (42%) and root length (61%) as compared with no tillage. In mungbean, tillage increased plant height (18%) and root length (60%), as averaged over both sites and seasons. In clay loam soil, tillage increased grain yield of maize by 242%. On the same field, tillage increased mungbean grain yield by 78% in 1984 and 20% in 1985. In sandy loam, tillage produced 38% more mungbean grains than without tillage.     

Influence of foliar application of manganese on growth,pigment content,and nitrate reductase activity of Vigna radiata (L.) R. Wilczek under salinity

The effect of the micronutrient manganese (Mn) on the physiological performance of green gram [Vigna radiata (L.) R. Wilczek] under saline condition was studied in the present work. Green gram was grown under controlled conditions and treated with different concentrations of sodium chloride (NaCl): 100, 200, and 300 mM. The plant samples were analyzed from 25 to 65 days of plant growth at every 10-day interval. Significant difference was observed in leaf area index, membrane stability index, nitrate reductase enzyme activity, total chlorophyll, and carotenoid content in treated plants. A drastic reduction was recorded at 200 and 300 mM NaCl treated plants in the parameters. The concentration 100 mM resulted in a slight increase of the parameters. Foliar application of Mn as manganese chloride (MnCl₂) was found to improve the physiological parameters in green gram at 200 and 300 mM salinity level and was useful in alleviating the detrimental effects of NaCl.     

Effect of Effective Microorganism Application on Crop Growth,Yield, and Nutrition in Vigna radiata (L.) Wilczek in Different Soil Amendment Systems

A pot experiment was conducted in heat-sterilized soil to evaluate the effect of effective microorganism (EM) application on growth, yield, and nutrient uptake in Vigna radiata (L.) Wilczek var. NIAB Mung 98 in different soil amendment systems. Pot soil was amended with farmyard manure (FYM), Trifolium alexanrinum L. crop residues (TCR), and half (½NPK) and recommended dose (NPK) of chemical fertilizers. The EM application significantly enhanced shoot and root biomass in TCR-amended soil. However, grain yield was significantly enhanced in FYM, TCR, and NPK amendments by 24%, 15%, and 84%, respectively, as a result of EM application. Effect of EM application on nutrient uptake was variable with respect to soil amendment and plant growth stage. In general, EM application enhanced plant nitrogen (N), phosphorus (P), and potassium (K) nutrition in organic amendments while its effect was either negative or insignificant in chemical fertilizer amendments. Effects of EM application on plant nutrient uptake were more pronounced at maturity than at flowering stage.     

Cadmium (Cd) and Lead (Pb) Induced Changes in Growth,Some Biochemical Attributes,and Mineral Accumulation in Two Cultivars of Mung Bean [Vigna radiata (L.) Wilczek]

Addition of cadmium, lead, and cadmium + lead (Cd, Pb, and Cd+Pb) to growth medium leads to a reduction in seed germination, length, fresh and dry biomass of shoots and roots, total chlorophyll, chlorophyll a, chlorophyll b, and soluble proteins in plants. This study was undertaken on two mungbean cultivars (MN-92 and MN-98). Application of these metals in the growth medium reduced the bioabsorption of calcium (Ca), magnesium (Mg), and potassium (K). Concentrations of all the metals were greater in the roots than in the shoots except for K, which was greater in the shoots. Mungbean cultivar NM-98 performed better than NM-92 in response to heavy-metal toxicity. Overall, the metal-induced physiobiochemical changes resulted in reduced growth of both mungbean cultivars.     

Comparison of symbiotic effectiveness of Rhizobium sp. with positive hydrogen-uptake activity compared with negative mutants in relation to nitrogen fixation in mungbean (Vigna radiata L.)

H₂ uptake activity was well distributed in Rhizobium sp. strains isolated from nodules of mung-bean (Vigna radiata L.). Two effective strains, RMP1 und RMP2, exhibiting significantly higher H₂ uptake activity were subjected to mutagenesis with nitrosoguanidine. The respective mutation frequencies were 0.18 and 0.19%. Three Hup^- mutants each of RMP1 und RMP2 were compared with the wild-type parent strains under pot culture experiments to evaluate the significance of the H₂ uptake system in biological N₂ fixation. Nodulation capabilities, plant growth characteristics, and the chlorophyll content of the leaves were significantly reduced in the plants treated with Hup^- mutants. Nitrogenase activity in Hup^- nodules was reduced by 8–41%. Similarly, N accumulation was also reduced singificantly.     

Root-mediated allelopathic interference of bhringraj (Eclipta alba L.) Hassk. on peanut (Arachis hypogaea) and mung bean (Vigna radiata)

The root mediated allelopathic interference of Eclipta alba infested soil on growth, physiological parameters and antioxidant enzyme activity was conducted on Arachis hypogaea L. and vigna radiata L. It was found that rhizosphere soil significantly reduced the germination percentage, seedling growth and dry biomass depending upon the species sensitivity. The germination inhibition was correlated with membrane deterioration as proved by a strong electrolyte leakage, increase in malondialdehyde (MDA) and H₂O₂ content. The physiological parameters like chlorophyll content, photosynthetic rate (P_n), intercellular CO₂ concentration (C_i), stomatal conductance (G_s), and transpiration (E) also showed significant reduction in E. alba infested soil and non-significant increase in leaf temperature (L_t) of two test species. The test seedlings have circumvented the allelochemicals stress, by both significant decrease and non-significant increase in the antioxidant activities in E. alba infested soil in contrast to control soil. Rhizosphere soil contained significantly higher amount of water-soluble phenolics as the putative allelochemicals, which were vanillic acid, benzoic acid, ferulic acid, and p-coumaric acid. The study concluded that rhizosphere soil exerts an allelopathic influence on peanut and mung bean by releasing water soluble phenolic acids as putative allelochemicals in soil.     

Potential of indigenous bradyrhizobia versus commercial inoculants to improve cowpea (Vigna unguiculata L. walp.) and green gram (Vigna radiata L. wilczek.) yields in Kenya

Limited information is available on reduced cowpea (Vigna unguiculata L. Walp.) and green gram (Vigna radiata L.Wilczek.) yields in Kenya. Declining soil fertility and absence or presence of ineffective indigenous rhizobia in soils are assumptions that have been formulated but still require to be demonstrated. In this study, soils were collected from legume growing areas of Western (Bungoma), Nyanza (Bondo), Eastern (Isiolo), Central (Meru) and Coast (Kilifi) provinces in Kenya to assess indigenous rhizobia in soils nodulating cowpea and green gram under greenhouse conditions. Our results showed that highest nodule fresh weights of 4.63 and 3.32?g plant^?1 for cowpea and green gram were observed in one soil from Isiolo and another from Kilifi, respectively, suggesting the presence of significant infective indigenous strains in both soils. On the other hand, the lowest nodule fresh weights of 2.17 and 0.72?g plant^?1 were observed in one soil from Bungoma for cowpea and green gram, respectively. Symbiotic nitrogen (N) fixation by cowpea and green gram was highest in Kilifi soil with values of 98% and 97%, respectively. A second greenhouse experiment was undertaken to evaluate the performance of commercial rhizobial inoculants with both legumes in Chonyi soil (also from Coast province) containing significant indigenous rhizobia [>13.5?×?10³ Colony Forming Units (CFU) g^?1]. Rhizobial inoculation did not significantly (P?相似文献   

Salt-induced perturbation in growth,physiological attributes,activities of antioxidant enzymes and organic solutes in mungbean (Vigna radiata L.) cultivars differing in salinity tolerance

A pot experiment was conducted to appraise the inhibitory effects of salt stress on biochemical attributes in the three mungbean cultivars (NCM-209, NCM-89 and NM-92). Salt stress caused a significant decrease in plant height, shoot relative water contents, photosynthetic pigments, endogenous levels of K⁺ and K⁺/Na⁺ ratios and increase in cellular levels of H₂O₂, MDA, Na⁺ and Cl^?. However, cv. NCM-209 was found to be tolerant in terms of lower salt-induced decline in K⁺, K⁺/Na⁺ ratio and photosynthetic pigments. The endogenous levels of H₂O₂ and MDA were also lower in cv. NCM-209. Salt stress markedly also affected different yield attributes in all mungbean cultivars. Again cultivar NCM-209 exhibited less inhibitory effects of salt stress on different growth attributes. Salt stress resulted in a marked increase in the activities of antioxidant enzymes (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase) in mungbean cultivars. Activity of peroxidase was maximal in cv. NCM-209 and catalase activity was maximal in cv. NCM-89, whereas cvs. NCM-89 and NM-92 showed higher activities of superoxide dismutase. Similarly activity of ascorbate peroxidase was higher in cv. NM-92. It could be inferred from data of antioxidant enzymes that mungbean cultivars cannot be categorized as salt tolerant or sensitive on the basis of a single antioxidant enzyme.