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[Genetic improvement of grain legumes of importance to Latin American diet and the soy bean].

Common beans and soybeans can be considered as the most important food legumes in the Latin American nutritional context, each of them presenting specific problems. The production of common beans has not increased in the last decade, and insects and diseases attacks are menacing it. On the other hand, soybeans have shown a tremendous increase in production, becoming rapidly an important export cash crop. However, its use in human consumption is quite restricted, in spite of the efforts of some countries in inducing their acceptance. As far as nutritive value is concerned, a high variability for protein and methionine contents have been evidenced, and breeding programs could use this factor advantageously. In the case of soybeans, the use of germ plasm with colored seeds appears to be a good alternative, due to the preference for color by the consumers. Considering the agricultural production policies of some of our countries, there is a need to establish a proper balance between the cash and energetic crops, and the staple food crops, in order to avoid an aggravation of the nutritional problems in this Continent.

Fabaceae↗

Effects of altitude above sea level on the cooking time and nutritional value of common beans.

The present study was conducted with the objective to determine the effects of altitude above sea level, on the cooking time and nutritional value of common black beans (Phaseolus vulgaris). Three 100 g samples of the Ostua variety were cooked at 8 individual locations, ranging in altitude from 0 to 2256 meters, in Guatemala, to establish water uptake and cooking time. The cooked samples were separated into cooked beans and cooking broth for chemical analysis. This included moisture, protein, lysine, tannins, total and enzyme susceptible starch, and fiber fractionation. The cooking liquor was analyzed for total solids, moisture, protein, ash and K. A 1200 g sample was cooked for the cooking time established previously, for biological testing of nutritional value, which included Net Protein Ratio (NPR), Protein Efficiency Ratio (PER), and protein digestibility. Altitude influenced cooking time which increased from 78 min at 0 m, to 264 min at 2256 m. Final moisture content in the cooked bean was similar at all altitudes and there was a tendency to yield smaller amounts of solids in the cooking broth at higher altitudes. The increase in cooking time was significant. Bean water uptake at all times was significantly slower and smaller at ambient as compared to water uptake at boiling T, at all altitudes. Protein and lysine content were not affected by altitude, however, tannin and catechin were lower in cooked samples, as compared to the raw material. Altitude did not affect the content of these substances. Total starch and total sugars were higher in the raw sample, as compared to the cooked samples, but there was no effect of altitude. Enzyme susceptible starch (ESS) was lower in the raw sample as compared to the cooked samples, which contained similar amounts with respect to altitude. No change was observed in fiber fractions of the cooked beans. Likewise, the composition of the cooking broth was very similar between cooking locations. There was a small tendency to a lower protein quality, with respect to altitude, the effect of which was more obvious in the apparent protein digestibility values. Undercooking or overcooking at one location influenced protein quality values. The extended cooking time of beans at high altitudes, has important economic and environmental implications, since significant amounts of wood have to be used.

Altitude↗

Characterization and comparison of arcelin seed protein variants from common bean.

Four variants of arcelin, an insecticidal seed storage protein of bean, Phaseolus vulgaris L., were investigated. Each variant (arcelin-1, -2, -3, and -4) was purified, and solubilities and M(r)s were determined. For arcelins-1, -2, and -4, the isoelectric points, hemagglutinating activities, immunological cross-reactivities, and N-terminal amino acid sequences were determined. On the basis of native and denatured M(r)s, the variants were classified as being composed of dimer protein (arcelin-2), tetramer protein (arcelins-3 and -4), or both dimer and tetramer proteins (arcelin-1). Although the dimer proteins (arcelins-1d and -2) could be distinguished by M(r)s and isoelectric points, they were identical for their first 37 N-terminal amino acids and had similar immunological cross-reactions, and bean lines containing these variants had a DNA restriction fragment in common. The tetramer proteins arcelin-1t and arcelin-4 also could be distinguished from each other based on M(r)s and isoelectric points; however, they had similar immunological cross-reactions and they were 77 to 93% identical for N-terminal amino acid composition. The similarities among arcelin variants, phytohemagglutinin, and a bean alpha-amylase inhibitor suggest that they are all encoded by related members of a lectin gene family.

Journal Article↗

The cytoplasmic male-sterility (CMS) determinant of common bean is widespread in Phaseolus coccineus L. and Phaseolus vulgaris L.

To identify regions of the mitochondrial genome that potentially could specify cytoplasmic male sterility (CMS) in Phaseolus coccineus (including P. polyanthus), and to define differences amongst P. coccineus lines, mitochondrial (mt)DNA restriction patterns and Southern blots of total DNA from sterile and fertile lines were analysed. By restriction endonuclease mapping we isolated a region which was specific to CMS lines flanking an F1-ATPase alpha-subunit (atpA) gene. DNA sequence analysis of this region showed 99.9% homology to the region previously isolated from P. vulgaris CMS Sprite. A high frequency of plants carrying the CMS-fragment was observed in a wild Phaseolus population, perhaps explaining the occurrence of inter- and intra-specific gene flow observed in the autogamous species P. vulgaris.

Blotting, Southern↗

Cotyledon thermal behavior and pectic solubility as related to cooking quality in common beans.

The characteristic of proteins, starch and pectic substances in cotyledons of two bean cultivars varying in cooking time were determined to investigate their possible contribution to bean cooking quality. Both cultivars showed the same enthalpies of starch gelatinization but different protein denaturation enthalpies. The proportion of hot water soluble pectins was higher in Michigan, the cultivar with the lower cooking time, than in Ojo de Cabra, the cultivar with the higher cooking time. These results were not due to differences in pectin methylation or in the ratio of monovalent to divalent cations in the tissue, suggesting that in fresh beans the beta-elimination reaction is not the sole or predominant route of thermal pectin degradation. Overall, this study indicates that varietal differences in bean cooking quality may be reflections of the rate of pectin loss during soaking/heating and that the thermal properties of starch and protein fractions seem to have a minor contribution. Researchers involved in this study propose that in fresh beans, the thermal pectin loss results from a two step mechanism: pectin enzymic breakdown during the bean soaking followed by thermal solubilization rather than beta-elimination during the bean heating.

Absorption↗

Ethiopian soils harbor natural populations of rhizobia that form symbioses with common bean ( Phaseolus vulgaris L.).

The diversity and taxonomic relationships of 83 bean-nodulating rhizobia indigenous to Ethiopian soils were characterized by PCR-RFLP of the internally transcribed spacer (ITS) region between the 16S and 23S rRNA genes, 16S rRNA gene sequence analysis, multilocus enzyme electrophoresis (MLEE), and amplified fragment-length polymorphism. The isolates fell into 13 distinct genotypes according to PCR-RFLP analysis of the ITS region. Based on MLEE, the majority of these genotypes (70%) was genetically related to the type strain of Rhizobium leguminosarum. However, from analysis of their 16S rRNA genes, the majority was placed with Rhizobium etli. Transfer and recombination of the 16S rRNA gene from presumptively introduced R. etli to local R. leguminosarum is a possible theory to explain these contrasting results. However, it seems unlikely that bean rhizobia originating from the Americas (or Europe) extensively colonized soils of Ethiopia because Rhizobium tropici, Rhizobium gallicum, and Rhizobium giardinii were not detected and only a single ineffective isolate of R. etli that originated from a remote location was identified. Therefore, Ethiopian R. leguminosarum may have acquired the determinants for nodulation of bean from a low number of introduced bean-nodulating rhizobia that either are poor competitors for nodulation of bean or that failed to survive in the Ethiopian environment. Furthermore, it may be concluded from the genetic data presented here that the evidence for separating R. leguminosarum and R. etli into two separate species is inconclusive.

DNA Fingerprinting↗

The role of shoot-localized processes in the mechanism of Zn efficiency in common bean.

Zn efficiency (ZE) is the ability of plants to maintain high yield under Zn-deficiency stress in the soil. Two bean ( Phaseolus vulgaris L.) genotypes that differed in ZE, Voyager (Zn-efficient) and Avanti (Zn-inefficient), were used for this investigation. Plants were grown under controlled-environment conditions in chelate-buffered nutrient solution where Zn(2+) activities were controlled at low (0.1 pM) or sufficient (150 pM) levels. To investigate the relative contribution of the root versus the shoot to ZE, observations of Zn-deficiency symptoms in reciprocal grafts of the two genotypes were made. After growth under low-Zn conditions, plants of nongrafted Avanti, self-grafted Avanti and reciprocal grafts that had the Avanti shoot scion exhibited Zn-deficiency symptoms. However nongrafted and self-grafted Voyager, as well as reciprocal grafts with the Voyager shoot scion, were healthy with no visible Zn-deficiency symptoms under the same growth conditions. More detailed investigations into putative shoot-localized ZE mechanisms involved determinations of leaf biomass production and Zn accumulation, measurements of subcellular Zn compartmentation, activities of two Zn-requiring enzymes, carbonic anhydrase and Cu/Zn-dependent superoxide dismutase (Co/ZnSOD), as well as the non-Zn-requiring enzyme nitrate reductase. There were no differences in shoot tissue Zn concentrations between the Zn-inefficient and Zn-efficient genotypes grown under the low-Zn conditions where differences in ZE were exhibited. Shoot Zn compartmentation was investigated using radiotracer ((65)Zn) efflux analysis and suggested that the Zn-efficient genotype maintains higher cytoplasmic Zn concentrations and less Zn in the leaf-cell vacuole, compared to leaves from the Zn-inefficient genotype under Zn deficiency. Analysis of Zn-requiring enzymes in bean leaves revealed that the Zn-efficient genotype maintains significantly higher levels of carbonic anhydrase and Cu/ZnSOD activity under Zn deficiency. While these data are not sufficient to allow us to determine the specific mechanisms underlying ZE, they certainly point to the shoot as a key site where ZE mechanisms are functioning, and could involve processes associated with Zn compartmentation and biochemical Zn utilization.

Adaptation, Physiological↗

The complete nucleotide sequence of a Pakistani isolate of Watermelon mosaic virus provides further insights into the taxonomic status in the Bean common mosaic virus subgroup.

Watermelon mosaic virus (WMV) is a potyvirus with a worldwide distribution, but is mostly found in temperate and Mediterranean regions. The complete nucleotide (nt) sequence of a Pakistani isolate of WMV (WMV-Pk) was determined and compared with French isolate (WMV-Fr) and other closely related potyviruses. WMV-Pk showed overall identities of 94.4% (nt) and 96% (amino acid; aa) with the WMV-Fr. However, variability was observed in the 5' UTR and P1 region. Although sequence identities over most of the genome were well above 90% at both the nt and aa levels, reaching 99.6% (aa) in the CP and 100% (aa) in the 6K1 and 6K2, thereby suggesting that WMV-Pk and WMV-Fr are identical strains, but the sequence identities in the P1 region were only 80.6% (aa) and 82.8% (nt), while that in the 5' UTR was 82%. These differences may be due to different mutation phenomena of a common ancestor virus or mutations caused by different selection pressures in two different agro-ecological zones. The sequence of WMV-Pk is very close to that of Soybean mosaic virus (SMV) over most of the genome, except for the N-terminal region, which is subject to recombination between SMV and Peanut stripe virus (PSV)/Bean common mosaic virus (BCMV), as revealed by Simplot and phylogenetic analyses of N- and C-terminal P1, HC-Pro, and 5' UTR regions of the genome.

Base Sequence↗

Dynamics of inositol phosphate pools (tris-, tetrakis- and pentakisphosphate) in relation to the rate of phytate synthesis during seed development in common bean (Phaseolus vulgaris).

Four cultivars of Phaseolus vulgaris were grown in a greenhouse and each flower was Labeled with date of anthesis. Seeds were collected at six different stages of development and inositol phosphates (InsPs) were analyzed by ion-pair reversed-phase HPLC. Phytate accumulation was similar in all cultivars, and the specific rate of phytate synthesis (Rs) peaked at about 22 days after flowering (DAF). Variations in the concentrations of the InsP3 and InsP4 pools matched changes in Rs in cultivars Una and Aruã. These results suggest mass-action effects. Thus, the rates of conversion of InsP3 to InsP5 appeared to be at least partly dependent on substrate concentration. Proportional increases in size of all InsP pools up to 21 DAF are also consistent with Little regulation in this part of the pathway. However, this did not appear to be the case in cv. Diamante Negro or with the conversion of InsP5 to InsP6 in all cultivars, where concentrations of the InsP precursor pools peaked earlier or even dropped as Rs peaked, suggesting activation of enzyme activity. Therefore, the evidence is consistent with a control point regulating this metabolic route upstream of InsP3 and possibly in the conversion of InsP5 to InsP6.

Flowers↗

Changes in ascorbate peroxidase, catalase, guaiacol peroxidase and superoxide dismutase activities in common bean (Phaseolus vulgaris) nodules under salt stress.

To analyse nodular antioxidant enzyme expression in response to salt stress, Phaseolus vulgaris genotype BAT477 was inoculated with reference strain CIAT899, and treated with 50 mM NaCl. Plant growth, nodulation and nitrogen fixing activity were analysed. Results showed that: (1) all parameters, particularly in nodules, were affected by salt treatments, and (2) confirmed preferential growth allocation to roots. The ARA was significantly decreased by salt treatments. Protein dosage confirmed that nodules were more affected by salt treatment than were roots. We analysed superoxide dismutase, catalase, ascorbate peroxidase and peroxidase in nodules, roots and a free rhizobial strain. Our results indicated that SOD and CAT nodular isozymes had bacterial and root origins. The SOD expressed the same CuZn, Fe and Mn SOD isoforms in nodules and roots, whereas in free rhizobia we found only one Fe and Mn SOD. APX and POX nodule and root profiles had only root origins, as no rhizobial band was detected. Under salt stress, plant growth, nitrogen fixation and activities of antioxidant defense enzymes in nodules were affected. Thus, these enzymes appear to preserve symbiosis from stress turned out that NaCl salinity lead to a differential regulation of distinct SOD and POX isoenzyme. So their levels in nodules appeared to be consistent with a symbiotic nitrogen fixing efficiency hypothesis, and they seem to function as the molecular mechanisms underlying the nodule response to salinity.

Ascorbate Peroxidases↗

Post-translational processing of two alpha-amylase inhibitors and an arcelin from the common bean, Phaseolus vulgaris.

Mass spectrometric methods were used to investigate the proteolytic processing and glycopeptide structures of three seed defensive proteins from Phaseolus vulgaris. The proteins were the alpha-amylase inhibitors alphaAI-1 and alphaAI-2 and arcelin-5, all of which are related to the seed lectins, PHA-E and PHA-L. The mass data showed that the proteolytic cleavage required for activation of the amylase inhibitors is followed by loss of the terminal Asn residue in alphaAI-1, and in all three proteins, seven or more residues were clipped from the C-termini, in the manner of the seed lectins. In most instances, individual glycoforms could be assigned at each Asn site, due to the unique masses of the plant glycopeptides. It was found that alphaAI-1 and alphaAI-2 differed significantly in their glycosylation patterns, despite their high sequence homology. These data complement the previous X-ray studies of the alpha1-amylase inhibitor and arcelin, where many of the C-terminal residues and glycopeptide residues could not be observed.

Amino Acid Sequence↗

Studies on the rhizosphere and rhizoplane microflora of common-bean and barley.

Both broad bean and barley plants stimulated bacteria (total), N-fixing clostridia, Azotobacter ammonifiers, and aerobic cellulose-decomposers in their rhizosphere zone. The rhizosphere effects were generally positive until the 8th week of either plant growth. However, the rhizosphere effect differed according to type of plant, growth phase of each single plant, and type of micro-organism under study. The presence of high numbers of micro-organisms in the rhizoplane samples (washed ruptured roots) of each plant supports the evidence that roots of these plants are colonized with soil micro-organisms.

Azotobacter↗

Protein structures of common bean (Phaseolus vulgaris) alpha-amylase inhibitors.

Two nucleotide sequences for genes that encode alpha-amylase inhibitor 4 (alphaAI-4) from white kidney bean (WKB) cv. 858, designated gene alphaAI-4 (Accession No. ), and alpha-amylase inhibitor 5 (alphaAI-5) from black bean (BB), designated gene alphaAI-5 (Accession No. ), were determined. Genes alphaAI-4 and alphaAI-5 encode 244 amino acid prepro-alphaAI-4 and prepro-alphaAI-5 polypeptides that are 93 and 95% identical with alpha-amylase inhibitor l (alphaAI-l; Hoffman, L. M.; Ma, Y.; Barker, R. F. Nucleic Acids Res. 1982, 10, 7819-7828), 40 and 43% identical with red kidney bean lectin, and 52 and 55% identical with arcelin l of wild-type bean. The high degree of sequence similarity indicates the evolutionary relationship among these genes. PCR analysis of genomic DNA purified from six genotypes of Phaseolus vulgaris showed very similar band patterns in 2% agarose gel, another indication of the conserved size homology among these genes. Proteolytic processing sites were located between Asn77 and Ser78 for pro-alphaAI-4 and pro-alphaAI-5. A bend next to Asn77 in three-dimensional model structures of alphaAI-4 and alphaAI-5 proinhibitors indicates that the proteolytic cleavage is necessary to remove the conformational constraint for activation to the mature protein. Mature WKB alphaAI-4 was composed of four subunits (2alpha2beta) and had a molecular weight of 50000 determined by multiangle laser light scattering and 56714 determined by laser-assisted time-of-flight mass spectrometry.

Amino Acid Sequence↗

Antioxidant activity in common beans (Phaseolus vulgaris L.).

Beans were pearled to evaluate the feasibility of increasing antioxidant activity and phenolic antioxidants. Phenolics were concentrated mostly in the hull fraction at about 56 mg of catechin equivalents per gram of sample. The methanolic extracts of the pearled bean samples were screened for antioxidant potential using the beta-carotene-linoleate and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) in vitro model systems. The pearled material, also referred to as milled samples, exhibited antioxidant activity that correlated with phenolic content and inhibited DPPH significantly in a dose-dependent manner. Phenolics and antioxidant activities were also examined in chromatographic fractions of methanolic extracts of manually obtained hulls that represented a model used previously to ascertain antimutagenic activity. Fractions extracted with ethyl acetate/acetone and acetone displayed antioxidant activity, which implies potent free radical scavenging activity with antimutagenic activity.

Acetates↗

Polyphenols in wild and weedy Mexican common beans (Phaseolus vulgaris L.).

The polyphenolic composition of 62 wild and weedy Mexican bean collections from diverse origins, grouped by their seed coat color, was assessed. According to spectrophotometric analysis, the range of total phenols, condensed tannins, and total anthocyanins presented wide differences. Furthermore, the phenolic acid, flavonoid, and anthocyanin profiles were analyzed using HPLC. Ferulic was the main phenolic acid. Kaempferol and quercetin were the main flavonoids, and the isoflavones daidzein and coumestrol were found in only low levels in few collections. Delphinidin was the main anthocyanidin found, followed by petunidin, cyanidin, malvidin, pelargonidin, and peonidin. The wide variation observed in polyphenolic contents was more related to their genotype than to the color factor. These results show that some wild and weedy beans are good sources of phenolic compounds for use in breeding programs focused on nutrition and health.

Anthocyanins↗