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PubMed2006

Biochemical composition and indigestible oligosaccharides in Phaseolus vulgaris L. seeds.

Common beans have a high nutritional value, but contain galactooligosaccharides (GO), which cause flatulence and intestinal discomfort in humans. The biochemical composition of ten bean cultivars was determined to select those of high protein and low GO contents. The cultivars varied in carbohydrate (47.02-60.17%), GO (3.12-5.71%), protein (22.17-33.50%), lipid (1.13-1.81%), moisture (11.42-12.93%) and ash contents (4.08-5.61%). 'Mexico 222' presented the highest alpha-galactosidase activity. Protein and GO contents were positively correlated. 'Perry Marrow' combined high protein and low GO concentrations, indicating it can be used in improvement programs aiming at high-quality cultivars for human consumption.

Dietary Proteins↗

PubMed1993

In vitro destabilization of plant viruses and cDNA synthesis.

DNA copies of a wide range of RNA viruses can be made by the direct addition of appropriately treated, purified virus particles to a reverse transcription reaction. Therefore, many problems associated with RNA isolation can be circumvented. Virus particles can be sufficiently destabilized by adjustments of salt content, buffer, pH or by the use of physical force supplied by a freeze/thaw cycle so that RNA in sufficient quantity and physical condition is available for the synthesis of in some cases, full length cDNAs. cDNAs have been made of viruses in the bromo-, poty-, carla-, ilar-, potex-, tobra and tobamovirus groups. Reported here are experiments with cowpea chlorotic mottle virus and bean common mosaic virus.

Blotting, Southern↗

PubMed2005

Influence of host cultivars and Rhizobium species on the growth and symbiotic performance of Phaseolus vulgaris under salt stress.

In order to study the effect of salt stress on the Rhizobium-common bean symbiosis, we investigated the response of both partners, separately and in symbiosis. The comparison of the behaviour of five cultivars of Phaseolus vulgaris differing in seed colour, growing on nitrates and different concentrations of NaCl, showed genotypic variation with respect to salt tolerance. Coco Blanc was the most sensitive cultivar, whereas SMV 29-21 was the most tolerant one. At the Rhizobium level, two strains previously selected for their salt tolerance were used: Rhizobium tropici strain RP163 and Rhizobium giardinii strain RP161. Their relative growth was moderately decreased at 250mM NaCl, but they were able to grow at a low rate in the presence of 342 mM NaCl. Their viability at the minimal inhibitory concentration was slightly affected. The effect of salinity on Rhizobium-plant association was studied by using the tolerant variety SMV 29-21 and the sensitive one Coco Blanc inoculated separately with both strains. In the absence of salinity, the strains induced a significantly higher number of nodules on the roots of the cultivar SMV 29-21 compared to those of Coco Blanc. Concerning effectiveness, both strains were similarly effective with SMV 29-21, but not with Coco Blanc. In the presence of salinity, Coco Blanc was more severely affected when associated with RP163 than with RP161. Salinity affected the nodulation development more than it affected the infection steps. Neither of the two strains was able to nodulate SMV 29-21 under saline conditions, in spite of the fact that this was considered the most salt-tolerant variety. The unsuccessful nodulation of SMV 29-21 could be related to the inhibition by salt of one or more steps of the early events of the infection process. In conclusion, N-fixing plants were found to be more sensitive to salt stress than those depending on mineral nitrogen. Evidence presented here suggests that a best symbiotic N2 fixation under salinity conditions could be achieved if both symbiotic partners, as well as the different steps of their interaction (early events, nodule formation, activity, etc.), are all tolerant to this stress.

Adaptation, Physiological↗

PubMed2004

Optimization modeling of plant root architecture for water and phosphorus acquisition.

An optimization model is presented that examines the relationship between root architecture and multiple resource acquisition, specifically water and phosphorus in spatially heterogeneous environments. The basal root growth angle of an individual common bean plant, which determines the orientation and localization of the bulk of the root system, was modeled as the decision variable. The total payoff to the plant, the benefit obtained from water and phosphorus acquisition, minus the costs of spatial competition between roots, is given as a function of the (x,y) coordinates of the basal root in two-dimensional Cartesian space. We obtained a general solution and applied it to four unique environmental cases which are as follows: (1) the case of uniformly distributed water and phosphorus; (2) the case of localized shallow phosphorus; (3) the case of localized deep water; and (4) the case of shallow phosphorus and deep water. The general solution states that the optimal basal root growth angle will occur at the point where the total rate of change in the value of the resources acquired equals the total rate of change in cost that results from locating the root deeper in the soil. An optimizing plant locates its roots deeper in the soil profile until the marginal benefit exactly equals the marginal cost. The model predicts that the basal root angle of an optimizing plant will be shallower for Case 2 and deeper for Case 3, relative to the basal root angle obtained in the case of uniformly distributed water and phosphorus. The optimal basal root angle for Case 4 will depend on the marginal rate of substitution of water availability for phosphorus availability that occurs with depth. Empirical observations of bean root architecture in the greenhouse and in the field confirm model results and are discussed. In addition, the potential importance of phenotypic plasticity and phenotypic variation are discussed in relation to optimization of traits and adaptation to spatially heterogeneous environments.

Adaptation, Physiological↗

PubMed2005

A ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil.

A direct molecular method for assessing the diversity of specific populations of rhizobia in soil, based on nested PCR amplification of 16S-23S ribosomal RNA gene (rDNA) intergenic spacer (IGS) sequences, was developed. Initial generic amplification of bacterial rDNA IGS sequences from soil DNA was followed by specific amplification of (1) sequences affiliated with Rhizobium leguminosarum "sensu lato" and (2) R. tropici. Using analysis of the amplified sequences in clone libraries obtained on the basis of soil DNA, this two-sided method was shown to be very specific for rhizobial subpopulations in soil. It was then further validated as a direct fingerprinting tool of the target rhizobia based on denaturing gradient gel electrophoresis (DGGE). The PCR-DGGE approach was applied to soils from fields in Brazil cultivated with common bean (Phaseolus vulgaris) under conventional or no-tillage practices. The community fingerprints obtained allowed the direct analysis of the respective rhizobial community structures in soil samples from the two contrasting agricultural practices. Data obtained with both primer sets revealed clustering of the community structures of the target rhizobial types along treatment. Moreover, the DGGE profiles obtained with the R. tropici primer set indicated that the abundance and diversity of these organisms were favoured under NT practices. These results suggest that the R. leguminosarum-as well as R. tropici-targeted IGS-based nested PCR and DGGE are useful tools for monitoring the effect of agricultural practices on these and related rhizobial subpopulations in soils.

Agriculture↗

PubMed2005

Inhibitory specificity and insecticidal selectivity of alpha-amylase inhibitor from Phaseolus vulgaris.

The primary structure and proteolytic processing of the alpha-amylase isoinhibitor alpha AI-1 from common bean (Phaseolus vulgaris cv. Magna) was determined by protein chemistry techniques. The inhibitory specificity of alphaAI-1 was screened with a panel of the digestive alpha-amylases from 30 species of insects, mites, gastropod, annelid worm, nematode and fungal phytopathogens with a focus on agricultural pests and important model species. This in vitro analysis showed a selective inhibition of alpha-amylases from three orders of insect (Coleoptera, Hymenoptera and Diptera) and an inhibition of alpha-amylases of the annelid worm. The inhibitory potential of alphaAI-1 against several alpha-amylases was found to be modulated by pH. To understand how alphaAI-1 discriminates among closely related alpha-amylases, the sequences of the alpha-amylases sensitive, respectively, insensitive to alphaAI-1 were compared, and the critical determinants were localized on the spatial alpha-amylase model. Based on the in vitro analysis of the inhibitory specificity of alphaAI-1, the in vivo activity of the ingested alphaAI-1 was demonstrated by suppression of the development of the insect larvae that expressed the sensitive digestive alpha-amylases. The first comprehensive mapping of alphaAI-1 specificity significantly broadens the spectrum of targets that can be regulated by alpha-amylase inhibitors of plant origin, and points to potential application of these protein insecticides in plant biotechnologies.

Amino Acid Sequence↗

PubMedDate not supplied

Partial purification and characterization of a non-specific acid phosphatase in leaves and root nodules of Phaseolus vulgaris.

Acid phosphatase (ACP) activity in common bean grown with or without 1.5 mM of phosphate has been examined. Leaves and root nodules responded to the absence of an exogenous phosphate source with an increase in ACP activity. Increases in enzyme activity were not associated with the synthesis of new isoforms of the enzyme. We partially purified and characterized the ACPs, which consisted of three proteins, one of leaf and two of nodule. Proteins of leaf migrated at 72 and 51 kDa in SDS-PAGE, whereas that of nodule migrated at 72, 49, 41 and 34 kDa. Enzymes of both organs had a pH optimum of 5.6, and were relatively heat stable. The enzymes exhibit a broad substrate selectivity, with maximal activity obtained with alpha-naphthyl-phosphate, ribulose 1,5-bisphosphate and p-nitrophenyl-phosphate (p-NPP). Potent inhibition by Zn2+, Hg2+, Cu2+, Pb2+, Al3+ and (MoO4)2- was observed.

Acid Phosphatase↗

PubMed2005

Antimicrobial and insecticidal protein isolated from seeds of Clitoria ternatea, a tropical forage legume.

The tropical forage legume Clitoria ternatea (L.) has important agronomic traits such as adaptation to a wide range of soil conditions and resistance to drought. It is resistant to a number of pathogens and pests. These important traits gave us reasons to look more closely at the plant. A highly basic small protein was purified from seeds of C. ternatea to homogeneity by using ultrafiltration with Centricon-3 membrane tubes and preparative granulated-bed isoelectric focusing (IEF). A single protein band was obtained on both sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and IEF gels. The protein, designated 'finotin', has broad and potent inhibitory effect on the growth of various important fungal pathogens of plants, namely Rhizoctonia solani, Fusarium solani, Colletotrichum lindemuthianum, Lasiodiplodia theobromae, Pyricularia grisea, Bipolaris oryzae and Colletotrichum gloeosporioides. It also inhibits the common bean bacterial blight pathogen Xanthomonas axonopodis pv. phaseoli. Moreover, finotin has powerful inhibitory properties against the bean bruchids Zabrotes subfasciatus and Acanthoscelides obtectus.

Animals↗

PubMed2005

Physiological implications of trehalase from Phaseolus vulgaris root nodules: partial purification and characterization.

The purification and characterization of trehalase from common bean nodules as well as the role of this enzyme on growth, nodulation nitrogen fixation by examining the effects of the trehalase inhibitor validamycin A, was studied. Validamycin A did not affect plant and nodule mass, neither root trehalase and nitrogenase activity; however this treatment applied at the time of sowing increased nodule number about 16% and decreased nodule trehalase activity (16-fold) and the size of nodules. These results suggest that nodule trehalase activity of Phaseolus vulgaris could be involved in nodule formation and development. In addition, acid trehalase (EC 3.2.1.28) was purified from root nodules by fractionating ammonium sulfate, column chromatography on DEAE-sepharose and sephacryl S-300, and finally on native polyacrylamide gel electrophoresis. The purified homogeneous preparation of native acid trehalase exhibited a molecular mass of 42 and 45 kDa on SDS-PAGE. The enzyme has the optimum pH 3.9, Km of 0.109 mM, Vmax of 3630 nkat mg-1 protein and is relatively heat stable. Besides trehalose, it shows maximal activity with sucrose and maltose and, to a lesser degree melibiose, cellobiose and raffinose, and it does not hydrolyze on lactose and turanose. Acid trehalase was activated by Na+, Mn2+, Mg2+, Li+, Co2+, K+ and inhibited by Fe3+, Hg+ and EDTA.

Cations↗

PubMed2005

Lotus japonicus: legume research in the fast lane.

Legumes are of immense importance to humanity and a key to sustainable agriculture. Two model species, Lotus japonicus and Medicago truncatula, are the focus of genome sequencing and functional genomics programmes, but most researchers focus exclusively on one or the other. In spite of this, legume researchers now have a unique opportunity to integrate work on these and other legume species, including soybean, common bean and pea to create a platform for comparative genomics second to none of any other plant family. The question is: do we have the scientific fortitude and political will to achieve this?

Cloning, Molecular↗

PubMed2006

Founder effect, plant host, and recombination shape the emergent population of begomoviruses that cause the tomato yellow leaf curl disease in the Mediterranean basin.

Tomato yellow leaf curl disease (TYLCD)-associated viruses present a highly structured population in the western Mediterranean basin, depending on host, geographical region and time. About 1,900 tomato and common bean samples were analyzed from which 111 isolates were characterized genetically based on a genome sequence that comprises coding and non-coding regions. Isolates of three distinct begomoviruses previously described were found (Tomato yellow leaf curl virus, TYLCV, Tomato yellow leaf curl Sardinia virus, TYLCSV, and Tomato yellow leaf curl Málaga virus, TYLCMalV), together with a novel recombinant virus. Mixed infections were detected in single plants, rationalizing the occurrence of recombinants. Except for TYLCV-type strain, single, undifferentiated subpopulations were present for each virus type, probably the result of founder effects. Limited genetic variation was observed in genomic regions, with selection against amino acid change in coding regions.

Begomovirus↗

PubMed2000

The effect of arcelin-1 on the structure of the midgut of bruchid larvae and immunolocalization of the arcelin protein.

Some wild accessions of the common bean (Phaseolus vulgaris) contain a family of proteins called arcelins, that are toxic to the larvae of certain bruchid species. Among the six allelic variants of arcelin tested so far, arcelin-5 and arcelin-1 confer the highest level of resistance against the Mexican bean weevil, Zabrotes subfasciatus. The same proteins are not toxic to the bean weevil, Acanthoscelides obtectus, which is also a serious pest of cultivated beans. Arcelins belong to the bean lectin family that includes phytohemaggutinins and alpha-amylase inhibitors. Although homologous to lectins, arcelins are themselves only very weak lectins, and their binding properties have not been clearly established. The toxic properties of arcelins may be related to their recognition of and interaction with the glycoproteins and other constituents of the membranes along the digestive tract of insects. Since arcelin-1 was shown to have growth inhibitory effects for the larvae of Z. subfasciatus but not of A. obtectus, we examined the effect of an arcelin-1 containing diet on the structure of the cells that line the intestinal tract of the larvae of these two bruchid species, and used antibodies against arcelin to examine the distribution of arcelin within the cells and tissues. Here we show that dietary arcelin-1 caused an alteration of the gut structure and the penetration of arcelin into the haemolymph in Z. subfasciatus but not in A. obtectus. These results lead us to suggest that arcelins exert their toxic effect by severely damaging the epithelial cells.

Journal Article↗

PubMed2001

Nitrate reductase activity, distribution, and response to nitrate in two contrasting Phaseolus species inoculated with Rhizobium spp.

The nitrate reductase activity distribution and response of two nodulated species of Phaseolus (Phaseolus vulgaris-common bean, and Phaseolus lunatus-lima bean) to different exogenous nitrate levels were studied during the vegetative period. These Phaseolus species showed to be very contrasting in respect to the pattern of nitrate reductase (NR) activity distribution thought the plant. The highest level of NR activity in P. vulgaris was clearly shown to occur in leaves in contrast with the lowest one detected in roots and nodules as widely seen for other tropical species of the Phaseoleae tribe. Conversely, P. lunatus had higher NR activity in the nodules, whereas its leaves exhibited a steadily decrease during the plant development. Indeed, at 32 days after emergence (pre-flowering stage), the nodulated P. vulgaris had approximately 95% of the total NR activity localized in its leaves, whereas in P. lunatus it was equally distributed in the nodules and in the leaves. Under long-term exposure to increasing exogenous level of nitrate, the leaf-NR activity of nodulated P. vulgaris presented a positive response, whereas the enzyme activity was very low and unresponsive in P. lunatus. In contrast, the nodule-NR activity showed a reverse response to the increasing NO(3)(-) level. The nodule-NR activity of P. lunatus significantly increased whereas in the P. vulgaris nodules it was very low and unresponsive. This present study suggests that P. lunatus inoculated with Rhizobium tropici presents a singular pattern of nitrate reduction distribution among leaves and nodules during the vegetative development. It is speculated that the nodulated Phaseolus lunatus may have different NR isoforms in their leaves (at least a constitutive type) and an inducible form in their nodules, responsive to long-term exposure to nitrate.

Journal Article↗

PubMed2000

A Phaseolus vulgaris lipoxygenase gene expressed in nodules and in Rhizobium tropici inoculated roots.

A genomic clone encoding a common bean lipoxygenase (PvLOX5) was isolated from a Phaseolus vulgaris library. Reverse transcription-polymerase chain reaction analysis revealed that PvLOX5 is expressed during nodule development and in Rhizobium tropici inoculated roots. There was no detectable expression of PvLOX5 in non-inoculated roots, healthy leaves, leaves after Pseudomonas syringae pv. tabaci infection, floral buds or dry seeds.

Amino Acid Sequence↗

PubMed2000

Molecular characterization of a new arcelin-5 gene.

Arcelins are insecticidal proteins found in some wild accessions of the common bean, Phaseolus vulgaris. They are grouped in six allelic variants and arcelin-5 is the variant with the highest inhibitory effect on the development of Zabrotes subfasciatus larvae. Characterization of the protein and its genes resulted in the identification of three polypeptides and the isolation of two genes that encode the Arc5a and Arc5b polypeptides. Here we describe a new gene, Arc5-III. The protein it encodes has 81% amino acid identity with the derived amino acid sequences of Arc5-I and Arc5-II. The Arc5-III gene is highly expressed in developing seeds and at a much lower level in roots. Data obtained by a combination of two-dimensional gel electrophoresis, protein sequencing and MALDI-TOF mass spectrometry analysis support the conclusion that Arc5-III encodes a polypeptide present in Arc5c band. Using ion-exchange chromatography, three fractions containing arcelin-5 polypeptides were eluted by increasing the salt concentration. The three fractions contain various amounts of the three arc-5 polypeptides and inhibit the growth of Zabrotes subfasciatus larvae differentially, suggesting differences in insecticidal activity among the arcelin-5 isoforms.

Amino Acid Sequence↗

PubMed1979

Influence of virus infection on the leaf protein yield.

Influence of cowpea mosaic virus (CPMV) and common bean mosaic virus (CBMV) infection on the yield of extractable leaf protein of cowpea cvs. Pusa Phaguni, Pusa Barsati, and Yard Long was studied. An increase as well as decrease in the yield of leaf protein concentrate (LPC), due to virus infection, has been observed. CPMV infection increased LPC yield in all the cowpea cvs., but CBMV decreased it in cys. Pusa Phaguni and Yard Long. Variation in percentage of extractable nitrogen and protein in LPC was also notices. However, total N and protein in leaf and LPC were increased due to virus infection, but total sugar and starch decreased.

Carbohydrates↗

PubMed2000

Alpha-amylases of the coffee berry borer (Hypothenemus hampei) and their inhibition by two plant amylase inhibitors.

The adult coffee berry borer (Hypothenemus hampei Ferrari [Coleoptera: Scolytidae]), a major insect pest of coffee, has two major digestive alpha-amylases that can be separated by isoelectric focusing. The alpha-amylase activity has a broad pH optimum between 4.0 and 7.0. Using pH indicators, the pH of the midgut was determined to be between 4.5 and 5.2. At pH 5.0, the coffee berry borer alpha-amylase activity is inhibited substantially (80%) by relatively low levels of the amylase inhibitor (alphaAI-1) from the common bean, Phaseolus vulgaris L., and much less so by the amylase inhibitor from Amaranthus. We used an in-gel zymogram assay to demonstrate that seed extracts can be screened to find suitable inhibitors of amylases. The prospect of using the genes that encode these inhibitors to make coffee resistant to the coffee berry borer via genetic engineering is discussed.

Animals↗

PubMed2003

Use of phage display to select novel cystatins specific for Acanthoscelides obtectus cysteine proteinases.

Cysteine proteinases from larvae of the common bean weevil, Acanthoscelides obtectus (Coleoptera: Bruchidae), were isolated by ion exchange affinity chromatography on a CM-Cellulose column and used to select mutant cystatins from a library made with the filamentous M13 phage display system. The library contained variant cystatins derived from the nematode Onchocerca volvulus cystatin through mutagenesis of loop 1, which contains the QVVAG motif that is involved in binding to proteinases. After three rounds of selection, the activity of variant cystatins against papain and cysteine proteinases from A. obtectus was assayed by ELISA. Two different variant cystatins (presenting amino acids DVVSA and NTSSA at positions 65-69) bound to A. obtectus cysteine proteinases more tightly than to papain. In contrast, the wild type had similar affinity for A. obtectus proteinases and for papain. These two selected variants cystatins have greater specificity towards A. obtectus cysteine proteinases than the original sequence and could represent good candidate genes for the production of transgenic plants resistant to this insect pest.

Animals↗