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Polymorphism of a complex resistance gene candidate family in wild populations of common bean (Phaseolus vulgaris) in Argentina: comparison with phenotypic resistance polymorphism.

Fifteen populations of wild bean (Phaseolus vulgaris), located in three provinces in Argentina, were analysed for their polymorphism for a complex resistance gene candidate (RGC) family clustered on the genome and for resistance phenotypes to strains of Colletotrichum lindemuthianum. Results indicate that RGC polymorphism is high. Population structure obtained for markers related to resistance was compared to population structure obtained for RAPD markers in order to infer the evolutionary forces driving polymorphism for resistance in wild populations at both molecular and phenotypic levels. Hierarchical analysis of differentiation showed that, within provinces, populations were differentiated for RAPD as well as for molecular and phenotypic markers of resistance. In contrast, provinces were differentiated only for RAPD and RGC markers and not for resistance phenotypes. The discrepancies found between diversity structures for molecular markers (RAPD and RGCs) and for resistance phenotypes suggest an effect of selection and indicate that diversity for resistance may not be driven by the same selective forces at the molecular and phenotypic levels. We further discuss whether specific selective forces are exerted on RGC markers and underline the importance of spatial scale of analysis for demonstrating an impact of selection.

Argentina↗

Characterization and gene expression of nodulin Npv30 from common bean.

We previously reported that transcripts for a 30-kD nodulin (Npv30) are very abundant in the nodule. In this paper we describe the isolation and characterization of Npv30 cDNA and genomic clones. Npv30 has the following characteristic features: (a) a putative signal sequence at the deduced amino-terminal region, (b) a proline-rich stretch at the carboxy terminus, and (c) a characteristic domain of four cysteines that resemble metal-binding sites. In Phaseolus vulgaris L., Npv30 is encoded by a small gene family that shares discrete sequence homologies with another small gene family in soybean. An antibody against a beta-galactosidase-Npv30 fusion protein detected two proteins of 28 and 30 kD. Although Npv30 transcripts are very abundant, they encode proteins that are hardly detected in nodule fractions, suggesting that these proteins have a short half-life and/or the mRNAs are strongly regulated at the translational level. Npv30 transcripts were detected in the infected cells of the nodule by in situ hybridization experiments.

Amino Acid Sequence↗

Characterization of the common bean uricase II and its expression in organs other than nodules.

Uricase II is a purine metabolic enzyme highly induced in root nodules during the symbiosis established between legumes and bacteria of the genera Rhizobium and Bradyrhizobium. Here we describe the characterization of bean (Phaseolus vulgaris) nodule uricase II cDNA and show that uricase II is encoded by a single gene in the bean genome. This gene is also expressed in cotyledons, roots, and hypocotyls during bean seedling establishment, and an anti-uricase antibody recognizes the protein in different seedling organs. Uricase II has also been found in Leucaena leucocephala seedlings, suggesting that it participates during seedling establishment in legumes that do not transport ureides. A 50-kD polypeptide that is detected by the anti-uricase antibody is found in cotyledons during seedling development. This higher-molecular-mass form is also detected in developing roots and hypocotyls but not in nodules. In situ hybridization experiments in root seedlings showed uricase II transcripts in the metaxylem parenchyma cells and phloem fibers of the vascular system.

Amino Acid Sequence↗

Anthocyanidins and Flavonols, Major nod Gene Inducers from Seeds of a Black-Seeded Common Bean (Phaseolus vulgaris L.).

Eleven compounds released from germinating seeds of a black-seeded bean (Phaseolus vulgaris L., cv PI165426CS) induce transcription of nod genes in Rhizobium leguminosarum biovar phaseoli. Aglycones from 10 of those compounds were identified by spectroscopic methods (ultraviolet/visible, proton nuclear magnetic resonance, and mass spectroscopy), and their biological activities were demonstrated by induction of beta-galactosidase activity in R. leguminosarum strains containing nodA-lacZ or nodC-lacZ fusions controlled by R. leguminosarum biovar phaseoli nodD genes. By making comparisons with authentic standards, the chemical structures for aglycones from the 10 molecules were confirmed as being anthocyanidins (delphinidin, petunidin, and malvidin) and flavonols (myricetin, quercetin, and kaempferol). All anthocyanidins and flavonols had 3-O-glycosylation and free hydroxyl groups at the 4', 5, and 7 positions. Hydrolysis experiments showed that the mean concentration required for half-maximum nod gene induction (I(50)) by the 10 glycosides was about half that of the corresponding aglycones. The mean I(50) value for the three anthocyanidins (360 nanomolar) was less (P </= 0.05) than that of the three flavonol aglycones (980 nanomolar). Each seed released approximately 2500 nanomoles of anthocyanidin and 450 nanomoles of flavonol nod gene inducers in conjugated forms during the first 6 hours of imbibition. Based on amounts and activities of the compounds released, anthocyanins contributed approximately 10-fold more total nod-inducing activity than flavonol glycosides. These anthocyanidins from bean seeds represent the first nod-inducing compounds identified from that group of flavonoids.

Journal Article↗

Rhizobium nod Gene Inducers Exuded Naturally from Roots of Common Bean (Phaseolus vulgaris L.).

Four compounds exuded from young roots of a black-seeded bean (Phaseolus vulgaris L., cv PI165426CS) induce transcription of nod genes in Rhizobium leguminosarum biovar phaseoli. The three most active nod gene inducers were identified by spectroscopic methods (ultraviolet/visible absorbance, proton nuclear magnetic resonance, and mass spectrometry) as being eriodictyol (5,7,3',4' -tetrahydroxyflavanone), naringenin (5,7,4' -trihydroxyflavanone), and a 7-O-glycoside of genistein (5,7,4' -trihydroxyisoflavone). Comparisons with authentic standards verified the chemical structures of the aglycones and their capacity to induce beta-galactosidase activity in R. leguminosarum strains containing nodA-lacZ or nodC-lacZ fusions controlled by R. leguminosarum biovar phaseoli nodD genes. Roots of 9-day-old seedlings released 42, 281, and 337 nanomoles per plant per day of genistein, eriodictyol, and naringenin, respectively. Genistein and naringenin induced higher maximum beta-galactosidase activities and required lower concentrations for half-maximum induction than eriodictyol. Comparing the nod gene-inducing activity of seed rinses with root exudate from PI165426CS bean showed that root flavonoids were released at about 6% the rate of those from seeds on a molar basis, but on average the individual compounds from roots were approximately three times more active than nod gene inducers from seeds.

Journal Article↗

Species limits in Rhizobium populations that nodulate the common bean (Phaseolus vulgaris).

Evolutionary genetic relationships among 146 bean-nodulating Rhizobium strains, including 94 field isolates from three localities in Colombia and 36 strains from Mexico, were examined by multilocus enzyme electrophoresis and restriction fragment length polymorphism analysis of a PCR-amplified 260-bp segment of the 16S rRNA gene. Seventy-five electrophoretic types (ETs), corresponding to multilocus enzyme genotypes, were identified, including a genotypically diverse group of 18 ETs in Colombia that is strongly differentiated from the ETs of R. etli, which occur in Mexico, Colombia, and Brazil. Most strains of the distinctive Colombian ETs carried the same 16S rRNA allele as did strains of R. etli, but, surprisingly, 17 isolates of two of these ETs had the allele that is characteristic of R. leguminosarum, and strains of two other divergent groups of ETs were also polymorphic for the two alleles. No fully satisfactory explanation for the occurrence of the R. leguminosarum 16S rRNA allele in three distantly related groups of strains is available, but horizontal transfer and recombination of the gene, in whole or in part, would seem to be more plausible than convergence in nucleotide sequence.

Alleles↗

Promotion of nod Gene Inducers and Nodulation in Common Bean (Phaseolus vulgaris) Roots Inoculated with Azospirillum brasilense Cd.

Inoculation of Phaseolus vulgaris with Azospirillum brasilense Cd promoted root hair formation in seedling roots and significantly increased total and upper nodule numbers at different concentrations of Rhizobium inoculum. In experiments carried out in a hydroponic system, A. brasilense caused an increase in the secretion of nod gene-inducing flavonoids, as was observed by nod gene induction assays of root exudates fractionated by high-performance liquid chromatography. Possible mechanisms involved in the influence of A. brasilense on this symbiotic system are discussed.

Journal Article↗

Mapping quantitative trait loci for a common bean (Phaseolus vulgaris L.) ideotype.

Breeding a model plant that encompasses individual traits thought to enhance yield potential, known as ideotype breeding, has traditionally focused on phenotypic selection of plants with desirable morphological traits. Broadening this breeding method to the molecular level through the use of molecular markers would avoid the environmental interactions associated with phenotypic selection. A population of 110 F5 recombinant inbred lines (RILs), derived from the cross between WO3391 and 'OAC Speedvale', was used to develop a genetic linkage map consisting of 105 random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), and sequence-tagged site (STS) markers. The map has a total length of 641 cM distributed across 8 linkage groups (LGs). Five of them were aligned on the core linkage map of bean. Twenty-one quantitative trait loci (QTLs) were identified over three environments for eight agronomic and architectural traits previously defined for a bean (Phaseolus vulgaris L.) ideotype. The QTLs were mapped to seven LGs with several regions containing QTLs for multiple traits. At least one QTL was located for each trait and a maximum of four were associated with lodging. Total explained phenotypic variance ranged from 10.6% for hypocotyl diameter to 45.4% for maturity. Some of the QTLs identified will be useful for early generation selection of tall, upright, high-yielding lines in a breeding program.

Analysis of Variance↗

Isolation and characterization of RNase LTR sequences of Ty1-copia retrotransposons in common bean (Phaseolus vulgaris L).

Retroelements have proved useful for molecular marker studies and play an important role in genome evolution. Ty1-copia retrotransposons are ubiquitous and heterogeneous in plant genomes, and although many elements have been isolated and characterized, almost no information about them is available in the literature for Phaseolus vulgaris L. We report here the isolation and characterization of new RNase long terminal repeat (LTR) sections of the Ty1-copia group for this crop plant. RNAse sections showed conserved amino acids with the downstream sections corresponding to the polypurine-tract and 5' sections of 3' LTRs. The RNase sections were aligned using ClustalX to find potential relationships between sequences. A comparison with this analysis was made using the partition analysis of quasispecies package (PAQ), which is specific for quasispecies-like populations. The analysis revealed eight distinct groups. To uncover LTR variability and potential conserved promoter motifs, we also designed new primers from the presumed polypurine-tract regions. A similarity search found short stretches similar to upstream and downstream regions of some genes. Conserved motifs, corresponding to transcription factor binding sites, were discovered through MatInspector software and two sequences characterized. From a putative LTR fragment, we then designed a new primer, which, through sequence-specific amplification polymorphism (SSAP), showed numerous polymorphic bands between two distinct P. vulgaris accessions.

Base Sequence↗

A genetic map of common bean to localize specific resistance genes against anthracnose.

A bean genetic map was developed to locate resistance genes against anthracnose and genes involved in plant defense mechanisms. One hundred and fifty-seven markers (51 restriction fragment length polymorphism, 100 random amplified polymorphic DNA, 2 sequence characterized amplified regions, and 4 morphological markers) were used to construct a genetic map covering 567.5 cM of the bean genome. Morphological markers consisted in two resistance genes towards anthracnose (Are and RVI), a dominant gene for nuclear male sterility (Ms8) and a pod-shape character (SGou). This map was established by using a backcross population (BC1) of 128 individuals, derived from a cross between two European bean genotypes: Ms8EO2 and Corel. Nine percent of the markers showed segregation distortions and mapped to three regions. Clusters of 2-10 markers were observed in every linkage group. The possible origin of these clusters is discussed. Nineteen markers shared with a previously published bean linkage map allowed us to establish a preliminary correspondence between the two maps. Finally, seven genes involved in plant defense mechanisms were located on this map.

Base Sequence↗

The dry plant extract of common bean seed (Phaseoli vulgari pericarpium) does not have an affect on postprandial glycemia in healthy human subject.

The aim of present study was to assess the effects and safety of a dry Phaseoli vulgari pericarpium (PVP) extract on postprandial glycemia in healthy participants. A randomized crossover experiment where participants received either PVP extract or placebo. Chemical compounds in dry extract were assessed by established methods. Eighteen healthy participants (9 male and 9 female) aged 29+/-4,8 years, body mass index (BMI) 23+/-3,7 kg/m(2) were recruited among students and staff at the Faculty of Pharmacy, University of Belgrade. All participants were able to follow the study protocol without difficulty. The participants received either PVP extract or placebo 30 minutes before a 50g oral glucose tolerance test (OGTT). The protocol followed the guidelines for the OGTT with blood samples drawn at 0, 15, 30, 60, 90 and 120 min. This study demonstrated that there was no significantly effect of the PVP extract on incremental blood glucose (IBG) and their areas under the curve (AUC) neither male nor female participants. However, IBG together with AUC changes were significantly lower in male compared with female participants in treated and untreated groups. The presence of chrome, soluble fiber, vitamin C, protein, glucose and lectins were also quantified. The applied amount of PVP extract was unable to produce the postprandial hypoglycemia. We assumed that amounts of chrome, soluble fiber, vitamin C which have beneficial effects on diabetes treatment were sufficient to produce hypoglycemia.

Adult↗

Phaseolin seed variability in common bean (Phaseolus vulgaris L.) by capillary gel electrophoresis.

Phaseolin, the major seed storage protein of Phaseolus vulgaris from forty-four wild and cultivated accessions, was studied using sodium dodecyl sulphate-capillary gel electrophoresis (SDS-CGE). In total, eleven phaseolin profiles, revealing polypeptide subunit variation in the range from 45.6 kDa to 54.4 kDa, were recorded. The number of polypeptide subunits recorded in particular profiles varied from 3 to 6; in total, eight phaseolin subunits were distinguished in the examined material. Ferguson plot analysis was used to correct non-ideal behaviour of phaseolin polypeptide subunits in capillary gel electrophoresis in the presence of SDS. The obtained results are compared to electrophoretic data received by slab polyacrylamide gel electrophoresis. The SDS-CGE method appears to provide a powerful tool for disclosure of phaseolin subunit variability.

Journal Article↗

[Effect of the seed coat on the hardening of common beans (Phaseolus vulgaris)].

Samples of cotyledons and whole black beans (Phaseolus vulgaris), Tamazulapa variety were stored during six weeks at 37 degrees C and 90% relative humidity to establish chemical and physical changes which occur during storage, and to determine the role played by the seed coat in the hard-to-cook phenomenon. After the storage period, samples of whole beans were divided in two subsamples, with and without the seed coat. These two samples and the cotyledons were analyzed for cooking time, water absorption, dietary fiber, tannic acid, soluble pectins and phytic acid. Cooking time of the whole beans increased from 99 to more than 480 minutes in the six-weeks period; for the cotyledons this value increased from 45 to 111 minutes. Cooking time of the dehulled bean, stored as whole bean, increased from 45 to 111 minutes. Cooking time of the dehulled bean, stored as whole bean, increased from 45 to 103 minutes. Water absorption in the whole beans and the cotyledons decreased, although in the cotyledons it was higher, due perhaps to the great absorption capacity to the seed coat. No changes were observed in the dietary fiber content of the cotyledons nor in the beans dehulled after storage. However, in the whole grains neutro-detergent fiber decreased, while acid detergent fiber, cellulose and lignin did not present significant changes. On the other hand, soluble pectates decreased in the whole bean and in the cotyledons; nevertheless the tannin content (as tannic acid) decreased only in the whole beans (from 3.28 to 1.64 mg/g). The data obtained suggest that the seed coat plays a significant role in the hard-to-cook process of hardening of the bean, before and during storage.

Dietary Fiber↗