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.     

Synthesis and resistance to in vitro proteolysis of transglutaminase cross-linked phaseolin, the major storage protein from Phaseolus vulgaris

The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin.     

Diversity of common bean (Phaseolus vulgaris L.) germplasm from Portugal

Common bean (Phaseolus vulgarisL.) is a traditional crop in Portugal, where farmers growvarieties selected and maintained by themselves. A collection of 88landraces of common bean was evaluated for 17 quantitative andqualitative traits and the biochemical marker phaseolin to displaythe degree of variation of this germplasm. Agronomic data weresubjected to cluster analysis and several groups were identified,with three groups clustering most of the landraces. Regardingphaseolin variation the C and T banding patterns are the mostfrequent ones, so the origin of the Portuguese beans is thus probablythe Andean region of South America. These results give informationabout the origin, diversity and breeding value of the Portuguesegermplasm, that could be useful to widen the genetic base ofcurrently cultivated bean varieties in Europe.     

Breeding for bean fly resistance in common bean (Phaseolus vulgaris L.): a review

The bean fly (Ophiomya spp) is the most important yield limiting insect pest of common bean in Africa. The insect pest can cause complete crop loss affecting bean production and productivity under epidemic conditions. Effective control of bean fly is essential for sustainable bean production Africa. The overall progress, opportunities and challenges of the bean fly control strategies. The biology and ecology of bean fly and the economic importance of the insect pest is presented as well as the existing controlling strategies, with an emphasis on the breeding on breeding strategies used, research progress achieved challenges and opportunities. In conclusion, significant research progress have been made in breeding for bean fly resistance evident by identification of breeding lines, understanding resistance mechanism and development of breeding strategies. However, there is a need for further research to validate the available information and also explore new breeding methods such molecular breeding which has not been explored at present. Such studies will accelerate breeding for bean fly resistance.     

Effect of nitrogen sources on the yield of common bean (Phaseolus vulgaris) in western Kenya

Depletion of soil nutrients due to continuous cultivation without adequate external fertilization is one of the challenges facing many smallholder farmers in western Kenya. This study was conducted to assess the effects of organic (water hyacinth compost), inorganic (urea) nitrogen (N) sources, and commercial Rhizobia inoculant on the yield of common bean (Phaseolus vulgaris) for two consecutive seasons in the short rains (2013) and long rains (2014). The experiments were laid out in a randomized complete block design and replicated four times. Triple superphosphate was applied to all treatments except those with compost to ensure that the soil had adequate phosphorus (P). Yellow and Rose coco bean varieties grown with urea and inoculated with commercial Rhizobia inoculant gave significantly higher yield of 382 kg ha^?1 and 341 kg ha^?1, respectively in the short rains (SR) season. In the long rains (LR) season bean yield was high in water hyacinth compost (1526 kg ha^?1) and control with non-limiting P (1300 kg ha^?1) treatments. Commercial Rhizobia inoculant did not significantly increase in yield in the SR and LR seasons. There was no significant influence on soil properties after two seasons of continuous cultivation of common bean and application of organic and inorganic fertilizers. These results demonstrate that water hyacinth compost improved bean yield in the LR season. However, longer field testing and economic analysis are required for it to be recommended as a substitute for inorganic N source among smallholder farmers.     

Composition of different bean varieties (Phaseolus vulgaris) of Northwestern Argentina (Region NOA): cultivation zone influence.

There have been prior investigations concerning the environmental effects and especially soil conditions upon the proximate analysis and mineral content of grains. However, the studies are not complete and have not involved beans grown in the northwestern regions of Argentina. For this reason, this study was initiated to determine the concentration of protein, moisture, ash, fat, and minerals of various bean samples grown in northwestern Argentina. Six varieties of beans were taken from seven different regions. AOAC standard methods were used for chemical analysis. The elements analyzed for all bean samples show that moisture varies from 12 to 14%, proteins from 18 to 22%, fat from 0.7 to 1.20%, copper from 0.8 to 1.20 mg/100 g, iron from 9 to 18 mg/100 g, zinc from 2.5 to 4 mg/100 g, and phosphorus from 295 to 542 mg/100 g. No arsenic was detected in the bean samples. Different analyzed bean varieties were significantly different for proximate composition and mineral content, and each variety from different regions of northwestern Argentina present significant differences.     

Proton release by nodulated roots varies among common bean genotypes (Phaseolus vulgaris) under phosphorus deficiency

Responses of proton release to phosphorus (P) availability by nodulated roots of common bean (Phaseolus vulgaris L.) were investigated for lines BAT 477 and CocoT, inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic culture under glasshouse conditions. Phosphorus was supplied as KH₂PO₄ at 15 and 60 μmol plant^–1 week^–1 (15P and 60P). Proton release was higher for BAT 477 than for CocoT under both P supplies. However, it was higher for 60P than 15P, whatever the line. The ratio of proton release per unit biomass of nodulated root was higher for BAT 477 than for CocoT, independent of P deficiency. Proton release was correlated with the nodulated‐root respiration for both genotypes and with the nodule respiration linked with nitrogen fixation for CocoT. Thus, the nodulation was more limited by 15P than root and shoot growth and more in CocoT than in BAT 477. It is concluded that independent of symbiotic N₂ fixation, proton release was higher in BAT 477 than in CocoT and that the nodulated legume releases a substantial amount of protons into its rhizosphere that is correlated with its nitrogen fixation that eventually depends upon the nodule permeability to O₂ diffusion.     

Resistance sources and reactions of common bean (Phaseolus vulgaris L.) cultivars in Turkey to anthracnose disease

Anthracnose caused by Colletotrichum lindemuthianum is one of the major diseases of common bean (Phaseolus vulgaris L.) worldwide. The aim of this study was to analyze common bean cultivars in Turkey for Co resistance genes using different molecular markers (SC08, SBB14, OPAZ04, SF10, OPF10, OPA18, SB12, TGA1, SAB3, SCAreoli, SZ04, and OPV20) linked with anthracnose resistance genes and evaluate the reaction of these cultivars to the pathogen isolates. The results showed the presence of Co-1, Co-1⁴, Co-1⁵, Co-2, Co-4, Co-4², Co-6, Co-10, Co-11, and Co-13 resistance genes, alone or combination except for Co-3³ and Co-5, in common bean cultivars. These cultivars carried predominantly Co-6 and Co-4 genes for anthracnose resistance. Five cultivar (Sazova, Zülbiye, Ak?n, Önceler and Karaca?ehir 90), carrying Co-1, Co-1⁴, Co-1⁵, Co-2, Co-10, Co-13, Co-4, and Co-6 resistance genes were resistant to the pathogen isolates. These cultivars, showing a high level of resistance to the pathogen can be used as resistance sources in common bean breeding programs for resistance to anthracnose disease. These markers will provide significant benefit in breeding strategies.

     

Biodiversity of common bean (Phaseolus vulgaris L.) in Honduras, evidenced by morphological characterization

This paper describes findings in the characterization of 300 accessions in a collection of common beans (Phaseolus vulgaris L.) in Honduras. The plant material was collected from 1990 to 1994 (4 years before the damaging Mitch Hurricane) covering most of the Departments of the country and kept at the Escuela Agrícola Panamericana Zamorano. Thirty-two morpho-agronomical characters were evaluated and the results were grouped by classes. The studied accessions showed a marked diversity with high predominance of red and small seeds. The accessions with purple flowers had black seeds and originate mainly from the western area of the country where this type of seed is preferred. Principal Component Analysis shows that clearly defined groups do not exist. The conservation of this diversity is recommended for future propagation, breeding and the investigation of the genetic relationships and other studies.     

Genetic diversity and population structure of a common bean (Phaseolus vulgaris L.) collection from Calabria (Italy)

Eighty-seven Phaseolus vulgaris landraces, still cultivated in Calabria (Italy), were investigated in order to study the patterns of common bean genetic diversity in this region, to better understand the evolutionary development of beans in Europe and to properly manage these genetic resources. Four American accessions and five Italian varieties were also included. Different markers, such as 12 microsatellites, seed traits, phaseolins and 100-seed weight were combined with different statistical approaches. For each microsatellite, expected (H _e ) and observed (H _o ) heterozygosities, polymorphism information content (PIC), probability of identity (PI) and homozygosity were calculated. Furthermore, in Calabrian group of bean landraces, total (N _a ) and private (N _pa ) number of alleles, observed (H _o ), expected heterozygosities (H _e ) and allelic richness (AR) were calculated. Genetic distances among landraces were estimated using Nei’s coefficient and a cluster analysis using the UPGMA algorithm was performed. The results clearly indicated that: (1) Calabrian germplasm showed a high level of diversity (H _e  = 0.595); (2) Mesoamerican and Andean gene pools were clearly distinguished in Calabrian germplasm, with the Andean gene pool predominating (83 %); (3) Calabrian landraces were largely hybridized within and between the gene pools. A model-based approach, using the STRUCTURE software, was adopted. Six groups, including 4 of Andean origin and one of Mesoamerican origin were identified. Even more interesting, a small group (8 %) showed a distinct genetic structure, in which interspecific hybridizations with runner bean (Phaseolus coccineus L.) could have occurred. Nevertheless, a relatively high proportion of Calabrian bean landraces (12.6 %) was derived from intra and interspecific hybridizations.     

Distribution and phenotypic characteristics of common bean (Phaseolus vulgaris L.) nodulating bacteria in diverse soils

ABSTRACT

Bacteria were isolated from the root nodules using common bean as a trap host. Growth and morphological characteristics of the bacterial isolates were described on yeast extract mannitol mineral salts agar and broth media. The results showed that over 67% of the isolates produced extracellular polysaccharide with an entire margin and convex elevation. The others differentiated into cream yellow, cream white and milky white colouration with colony area ranging from 0.8 to 26?mm². The population levels of bacteria nodulating common bean varied in soils and were not affected by the cropping system. However, selected soil parameters greatly influenced the occurrence and distribution of these bacteria. The isolation of indigenous bacteria in all the soils with different cropping systems is an indication that the soils are favourable for nitrogen fixation. Based on the differences in cultural characteristics of the isolates, our data demonstrate the presence of high diversity of bacteria associated with bean nodules.     

Primers and probes development for specific PCR detection of genetically modified common bean (Phaseolus vulgaris) Embrapa 5.1

The genetically modified common bean Embrapa 5.1, developed by Brazilian Agricultural Research Corporation (Embrapa), is the first commercial GM plant produced in Latin America. It presents high resistance to the Bean golden mosaic virus. In this work, primers and probes targeting a taxon-specific reference DNA sequence for the common bean (Phaseolus vulgaris L.) and a construct-specific DNA sequence of Embrapa 5.1 GM common bean were successfully developed. The primers and probes showed high specificity for the target detection. Both methods showed suitable efficiency and performance to be used as an endogenous target for detection of common bean DNA and for construct-specific detection of GM common bean Embrapa 5.1, respectively. Both real-time PCR assays proved to be valuable for future assessment of interlaboratory studies.     

Effect of exudate released from seeds and seedling roots of common bean (Phaseolus vulgaris L.) on proliferation of Rhizobium sp. (Phaseolus)

Effects of seed and root exudates obtained from common bean on the proliferation of Rhizobium sp. (Phaseolus) were examined in a combination of three plant cultivars with three Rhizobium strains. In the first experiment, seed or root exudate was mixed with an Andosol soil extract, and bacterial proliferation in the mixture was traced. Seed exudate was prepared from hydroponic solution used in seed imbibition for 24 h, and a series of root exudates was prepared from a hydroponic solution collected every 24 h from the initiation of rooting up to 96 h after rooting. Regardless of the common bean cultivars and Rhizobium strains used, Rhizobium population markedly increased of the 24 h of culture in the mixture containing seed exudates, whereas a negligible increase was detected in the mixture with root exudates. The mixture containing root exudates collected within a period of 72–96 h after initial rooting (96–120 h after seed imbibition) exerted an inhibitory effect on Rhizobium proliferation. The seed exudates contained large amounts of sugars, amino acids, nitrogen, phosphorus, potassium, and magnesium compared to any root exudates. In the second experiment, Rhizobium was inoculated directly to common bean seeds sowed in a vermiculite bed which was sterilized and moistened with a plant nutrient solution. Compared with the control (without seed), a remarkable increase in the number of bacterial cells was observed in all the combinations of plant cultivars and Rhizobium strains 24 h after sowing. These results reveal that seed exudates of common bean have a substantial potential to promote Rhizobium proliferation, and that root exudates in a particular period of culture contain some inhibitory factors.     

Compatibility of crosses between gene pools and evolutionary classes in the common bean (Phaseolus vulgaris L.)

The extent of genetic divergence between the Mesoamerican and Andean gene pools and between evolutionary classes (wild, landrace and bred) in Phaseolus vulgaris was explored. The compatibility of crosses was assessed by the frequency with which pollination was successful and the number of seeds in the resulting pod. Mesoamerican genotypes were more effective as pollinators than Andean genotypes, regardless of the gene pool of the female parent. Though certain individual genotypes were more easily pollinated by genotypes of one or other of the gene pools, there was no consistent tendency for pollinations between genotypes within a gene pool to be more successful than those between gene pools. In crosses among wild and landrace genotypes, the landrace genotypes were more easily pollinated. In those among wild and bred genotypes, the bred genotypes were more effective as pollinators, and the number of seeds per pod was greater in crosses between the evolutionary classes than in those within classes. In crosses among landrace and bred genotypes, those within evolutionary classes had higher success rates than those between classes (47.2% vs. 38.9%), and produced more seeds per pod. However, these results do not represent a major barrier between the evolutionary classes. The hypothesis that dwarf-lethal incompatibility in inter-gene pool crosses is controlled by two complementary dominant genes (the DL-gene hypothesis) was tested by comparing the numbers of normal and abnormal plants in the progeny of three-way crosses, and the ratios obtained were consistent with the hypothesis. It is concluded that there is no biological barrier between the two gene pools and that they are distinct because, by accident of ancestry, they are located in geographically isolated and ecologically distinct areas, with different cultural practices and preferences. Research programmes aimed at transferring desirable traits between the gene pools should therefore be encouraged.     

Iron bioavailability to piglets from red and white common beans (Phaseolus vulgaris)

Polyphenols in foods may chelate dietary Fe and lower its bioavailability. Concentrations of phenols are higher in red beans than in white beans. The aim of this study was to compare iron bioavailabilities from red and white beans in a piglet hemoglobin repletion model. Fe deficient cross bred piglets (Hampshire x Landrace x Yorkshire) were used. Nutritionally balanced diets (except for Fe) were formulated to contain 50% precooked, dehydrated beans (either small red or Great Northern white). At age 5 weeks, the piglets were assigned to two groups and fed diets containing either red or white beans for 4 weeks. Weight and hemoglobin (Hb) concentrations were monitored weekly. Feed intakes were measured daily. Hemoglobin repletion efficiency (HRE) was calculated as the gain in total body hemoglobin Fe (Hb-Fe) divided by Fe intake. Hb concentrations, Hb-Fe gains, and HRE were not different between the groups at any time point ( p > 0.05). HRE values in the red bean group were 50% in the first week and 30% over the entire 4 week period. In the white bean group, they were 56 and 26%, respectively. Proline-rich protein mRNA concentrations in parotid glands were higher in the red bean group compared to the white bean group. These results show that iron bioavailabilities from red and white beans are similar and suggest that pigs adapt to the inhibitory effects of polyphenols on iron absorption by increasing the secretion of protective proline-rich proteins in the saliva.     

Chemical components with health implications in wild and cultivated Mexican common bean seeds (Phaseolus vulgaris L.)

Common bean effects on health have been related to its dietary fiber content and other active compounds. This study assessed the content of flavonoids, coumestrol, phenolic acids, galactooligosaccharides, and phytic acid in wild and cultivated Mexican common bean seeds (raw and cooked) and that of flavonoids, coumestrol, and phenolic acids in germinated bean seeds. The presence of isoflavones in raw bean seeds was not confirmed by the UV spectra. Quercetin, kaempferol, p-coumaric acid, ferulic acid, p-hydroxybenzoic acid, and vanillic acid mean contents were 10.9, 52.3, 10.1, 9.6, 5.4, and 18.2 microg/g, respectively; raffinose, stachyose, verbascose, and phytic acid mean contents were 8.5, 56.3, 5.5, and 11.5 mg/g, respectively, in raw seeds. All compounds were affected by autoclaving, and germination resulted in a de novo synthesis of flavonols, phytoestrogens, and phenolic acids. The impact on health of common bean seed is affected by dietary burden, specific compounds content, and processing. On the other hand, germinated bean seed or beans sprouts may be sources of antioxidants and phytoestrogens.     

Population structure and genetic differentiation among the USDA common bean (Phaseolus vulgaris L.) core collection

Genetic diversity data were collected from a large population of common bean (Phaseolus vulgaris L.) landraces representing the United States Department of Agriculture core collection. The data were based on microsatellite data from all linkage groups. A procedure was developed to determine if we collected sufficient marker data to adequately estimated pairwise diversity. The diversity data were used to define populations using distance and model-based approaches. Genetic differentiation and genetic isolation by distance data were collected. Diversity was also compared for markers linked and unlinked to domestication loci. Using a model-based approach, the landraces were divided into the traditional Middle American and Andean gene pools. Diversity was greater for the Middle American gene pool. Six Middle American and three Andean subpopulations were defined, and the Middle American subpopulations exhibited strong geographic identity. Unlike other studies, seed size varied considerably with subpopulations, and a number of the subpopulations contained landraces from multiple common bean races. All of the subpopulations were highly differentiated, with the Middle American subpopulations showing the greatest differentiation. Genetic isolation by distance was observed among the Middle American and Andean subpopulations but not among subpopulations within a gene pool. Within each gene pool, diversity was lower for markers linked to domestication loci.     

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.     

Biotransformation of common bean (Phaseolus vulgaris L.) flavonoid glycosides by bifidobacterium species from human intestinal origin

Bifidobacteria strains from human origin were screened for the specific activity (beta-glucosidase activity) involved in the metabolism of dietary flavonoids. Five strains with high beta-glucosidase activity were selected for further metabolism analyses (high-performance liquid chromatography separations) of flavonoid glycosides occurring in Phaseolus vulgaris L. (common bean) seeds and seedlings. All selected strains were found to be active in the conversion of kaempferol 3-O-glucoside, daidzin, genistin, and glycitin into their aglyconic forms. No metabolites were detected after the fermentation tests with the diglucosidic compound kaempferol 3-O-xylosylglucoside. In addition, to verify the effective bioavailability of flavonoid aglycones, the degradation rates of daidzein, genistein, glycitein, and kaempferol, following incubation with selected strains, were monitored. The results showed that the five selected strains of bifidobacteria, being active in the biotranformation of flavonoid glycosides occurring in common bean seeds and seedlings, could be considered as probiotic dietary adjuncts to improve the nutritional and health properties of flavonoid-based products, comprising hypothetical common bean food derivatives.