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PubMed1998

Stoichiometric shifts in the common bean mitochondrial genome leading to male sterility and spontaneous reversion to fertility

The plant mitochondrial genome is characterized by a complex, multipartite structure. In cytoplasmic male-sterile (CMS) common bean, the sterility-inducing mitochondrial configuration maps as three autonomous DNA molecules, one containing the sterility-associated sequence pvs-or f 239. We constructed a physical map of the mitochondrial genome from the direct progenitors to the CMS cytoplasm and have shown that it maps as a single, circular master configuration. With long-exposure autoradiography of DNA gel blots and polymerase chain reaction analysis, we demonstrate that the three-molecule CMS-associated configuration was present at unusually low copy number within the progenitor genome and that the progenitor form was present substoichiometrically within the genome of the CMS line. Furthermore, upon spontaneous reversion to fertility, the progenitor genomic configuration as well as the molecule containing the pvs-or f 239 sterility-associated sequence were both maintained at substoichiometric levels within the revertant genome. In vitro mitochondrial incubation results demonstrated that the genomic shift of the pvs-or f 239-containing molecule to substoichiometric levels upon spontaneous reversion was a reversible phenomenon. Moreover, we demonstrate that substoichiometric forms, apparently silent with regard to gene expression, are transcriptionally and translationally active once amplified. Thus, copy number suppression may serve as an effective means of regulating gene expression in plant mitochondria.

Journal Article↗
PubMed1998

A stomatin-like protein encoded by the slp gene of Rhizobium etli is required for nodulation competitiveness on the common bean.

Rhizobium etli strain TAL182 is a competitive strain for effective nodulation of beans. From this strain, a novel gene was isolated, slp, which is 669 bp in size and required for nodulation competition on the common bean. The slp knockout mutant of TAL182 is defective in nodulation competition, shows reduced growth in the presence of 200 mM NaCl, KCl or LiCl and is complemented by the cloned slp gene. The deduced amino acid sequence of slp shows 66-72% similarity to stomatin proteins of Homo sapiens, Mus musculus and Caenorhabditis elegans. Expression of slp in Escherichia coli from a T7 promoter shows a 26 kDa protein which cross-reacts with human-stomatin-specific polyclonal antibody. Like the human stomatin protein, the slp-deduced protein, Slp, is very hydrophilic except for a single hydrophobic membrane-spanning domain. Among various bean-nodulating rhizobia, slp is present in R. etli, Rhizobium leguminosarum bv. phaseoli and Rhizobium tropici type A strains but is absent in R. tropici type B strains. It is also absent in Bradyrhizobium and several other Rhizobium spp.

Amino Acid Sequence↗
PubMed2000

Efficient whole plant regeneration of common bean (Phaseolus vulgaris L.) using thin-cell-layer culture and silver nitrate.

A method was designed to optimize rapid and high frequency direct shoot regeneration (without intermediate callus) of the commercially important common bean, Phaseolus vulgaris L., using the transverse thin cell layer (tTCL) method. The pretreatment of seeds with 10 µM TDZ significantly increased bud regeneration frequency on tTCL. A 2-week culture of tTCLs on 10 µM TDZ followed by a reduction in the TDZ concentration (1 µM) was needed to achieve optimal bud induction and further development of the neo-formed buds. An incubation period greater than 2 weeks of tTCLs with 10 µM TDZ concentration resulted in inhibitory effects on the development of shoots and roots. Shoot development was enhanced by 10 µM BAP and 10 µM AgNO(3) leading to 100% well developed shoots. Regenerated plants developed into true-to-type fertile plants.

Journal Article↗
PubMed2003

Effect of cultivar and growing location on the trypsin inhibitors, tannins, and lectins of common beans (Phaseolus vulgaris L.) grown in the semiarid highlands of Mexico.

Trypsin inhibitors (TI), tannins, and lectins appear to have a role in preventing chronic diseases in humans. The genetic variability of these traits in common bean needs to be ascertained in order to increase levels through breeding. The variability of TI, tannin, and lectins was determined in five bean cultivars grown at five locations in Mexico. TI and tannins contents in colored beans that belong to the Jalisco race were higher (11.1-11.9 trypsin units inhibited (TUI)/mg and 29.0-38.1 mg catechin equivalent (CE)/g, respectively) than cultivars of the Durango race (7.9-8.3 TUI/mg and 16.8-19.9 CE/mg, respectively). Bayo Victoria, a Durango race cultivar, had three times more lectins than levels reported for soybean. Cultivar influenced TI and tannins contents (p < 0.001), whereas site affected lectins (p < 0.001). An increase in levels of TI and tannins could be enhanced through breeding.

Lectins↗
PubMed2001

An interdisciplinary research strategy to improve symbiotic nitrogen fixation and yield of common bean (Phaseolus vulgaris) in salinised areas of the Mediterranean basin.

The main findings of a cooperative research group of agronomists, plant breeders, microbiologists, physiologists and molecularists to improve the symbiotic nitrogen fixation (SNF) and N2-dependent yield of common bean under moderate salinity in the Mediterranean basin are summarised. Agronomic surveys in reference production areas show large spatial and temporal variations in plant nodulation and growth, and in efficiency of utilisation of the rhizobial symbiosis. The latter was associated with a large rhizobial diversity, including new bean nodulating species. Macrosymbiont diversity in SNF and adaptation to NaCl was found. However, contrasts between plant genotypes could be altered by specific interactions with some native rhizobia. Therefore, variations in soil rhizobial population, in addition to agronomic practices and environmental constraints, may have contributed to erratic results observed in field inoculations. At the mechanistic level, nodule C and N metabolisms, and abcissic acid content, were related to SNF potential and tolerance to NaCl. Their relation with nodule conductance to O2 diffusion was addressed by in situ hybridisation of candidate carbonic anhydrase and aquaporin genes in nodule cortex. The limits and prospects of the cooperative strategy are discussed.

Gene Expression Regulation↗
PubMed1998

Sequences of the coat protein gene of five peanut stripe virus (PStV) strains from Thailand and their evolutionary relationship with other bean common mosaic virus sequences.

The coat protein gene and part of the 3' non-coding region of five strains of peanut stripe virus (PStV) from Thailand have been cloned and sequenced. Phylogenetic comparisons of these strains, known as T1, T3, T5, T6 and T7, and related sequences showed that these strains are indeed strains of PStV. Further, PStV strains appear to be related to each other according to their geographic origin. That is, the Thai strains are more closely related to each other than they are to strains from the USA or Indonesia, despite the variety of symptoms caused by these strains and the overlap of symptom types between the strains from different locations. Like other PStV strains, PStV-Thai can be considered strains of bean common mosaic virus (BCMV) but can be distinguished from bean-infecting strains of BCMV and blackeye cowpea mosaic virus (B1CMV) through sequence and host range. No evidence was found that PStV-Thai strains, unlike PStV-Ib, are recombinants of PStV and B1CMV, although the T3 strain may be a recombinant of different PStV sequences. Phylogenetic analyses of viruses of the BCMV group suggest that acquisition of the ability to infect peanut may have occurred only once.

Amino Acid Sequence↗
PubMed2000

Analysis of bruchid resistance in the wild common bean accession G02771: no evidence for insecticidal activity of arcelin 5.

Arcelins are abundant seed storage proteins thought to be implicated in the resistance of wild Phaseolus vulgaris (L.) genotypes against Zabrotes subfasciatus (Boheman), an important storage insect pest of common bean. Here, the insecticidal activity of the arcelin-5 variant that is present in the highly resistant P. vulgaris accession G02771 was investigated. No correlation could be established between the presence of arcelin 5 and the insecticidal effects observed in G02771 seeds. Insect feeding assays with artificial seeds into which purified arcelin-5 protein was incorporated and with transgenic P. acutifolius (A. Gray) seeds in which the arcelin-5 genes were expressed, showed that the presence of arcelin-5 proteins, even at elevated levels, was not sufficient to achieve adequate resistance against Z. subfasciatus. The same might apply to other arcelin variants. Nevertheless, as resistance is clearly closely linked to the presence of the arcelin-1 or arcelin-5 locus, arcelins remain useful markers in breeding programmes aimed at introgressing high levels of resistance to Z. subfasciatus in P. vulgaris cultivars.

Animals↗
PubMedDate not supplied

[Abscisic acid level, activities of proteinases and trypsin inhibitory proteins in the germinating seeds of common beans under conditions of water stress].

Specific features of changes in the contents of free and bound abscisic acid (ABA) and activities of neutral and alkaline proteinases and trypsin inhibitory proteins were determined in the embryonic axis and cotyledons of the common bean (Phaseolus vulgaris L.) after drying. The changes in ABA content, observed following the loss of 5% seed weight, were regarded as an adaptive reaction to stress, whereas the corresponding changes after the loss of 10% seed weight, as a result of pathological disturbance of ABA metabolism. Both drying modes had a negative effect on the state of the proteinase-inhibitory system, as was apparent from the disruption of the regular inverse correlation between the activities of proteinases and serine proteinase inhibitory proteins. Comparison of the dynamics of these characteristics with the buildup of water stress demonstrated an inverse correlation between the content of free ABA and the activity of the proteinases studied. This suggests a potential inhibitory effect of this hormone on the function of the hydrolases in question in the germinating seed.

Abscisic Acid↗
PubMed2002

Antimutagenic activity of natural phenolic compounds present in the common bean (Phaseolus vulgaris) against aflatoxin B1.

Polyphenols with antimutagenic and anticarcinogenic properties are present in fruits, vegetables and legumes. In this study, the Salmonella typhimurium tester strains TA98 and TA100 were used in the microsuspension assay to examine the antimutagenic effect of phenolic compounds extracted from the common bean (Phaseolus vulgaris) against mutagenicity induced by aflatoxin B1 (AFB1). A dose-response curve was constructed for AFB1; from which a level of 40 ng AFB1/tube was selected for all antimutagenicity assays. The AFB1 and phenolic extract (PE) were not toxic to the bacteria at concentrations tested. In the case of PE, results were similar to the number of spontaneous revertants for TA98 and TA100. The inhibitory effect of PE against AFB1 mutagenicity was dose-dependent at the lower concentrations tested (2.5, 5, 10, 12.5, 15 and 25 microgram-equivalent (+)-catechin/tube for TA98; 0.5, 1, 1.5, 2.5, 5, 10 and 25 microgram-equivalent (+)-catechin/ tube for TA100). Further, a two-stage incubation procedure was used to investigate the potential interaction between PE and AFB1. The greatest inhibitory effect of the PE on AFB1 mutagenicity occurred when PE and AFB1 were incubated together. When the bacteria were first incubated with PE followed by a second incubation with AFB1, lower inhibition was observed. Lower inhibition was also observed when the bacteria were first incubated with AFB1 followed by a second incubation with PE. The results suggest that the mechanism of inhibition could involve the formation of a chemical complex between of PE and AFB1.

Aflatoxin B1↗
PubMed1999

Analysis of lipoxygenase mRNA accumulation in the common bean (Phaseolus vulgaris L.) during development and under stress conditions.

Plant lipoxygenases (LOX, EC 1.13.11.12) have been involved in processes such as stress responses and development. The levels of these enzymes and their corresponding mRNAs are modulated during these processes as well as by different effectors such as jasmonic acid (JA), its methyl ester (MeJA) or abscisic acid (ABA). A new lipoxygenase (LOX) cDNA clone, PvLOX2, was isolated from a Phaseolus vulgaris nodule library and used to study the LOX mRNA accumulation pattern in some developmental stages and in plants subjected to hormone and stress treatments. In nodules, LOX mRNA reaches a maximum level around day 14 to 16 after Rhizobium tropici inoculation, as compared with LOX mRNA present in uninoculated and inoculated roots at the same days. LOX antigen is detected in the nodule parenchyma and in the uninfected cells. During germination, bean LOX transcripts start to accumulate 48 h after imbibition, remains at the same level until 72 h after imbibition and then declines. In hypocotyl, LOX mRNA is abundant in the growing region and almost absent in the mature region. After water stress or ABA treatment, this mRNA increases in the mature region and decreases in the growing region. In bean seedlings, LOX mRNA is accumulated in response to some types of stresses such as cold and desiccation. Wounding, MeJA or ABA treatment of mature leaves also induces LOX mRNA accumulation. These results indicate that in common bean plants LOX is required during development and stress conditions.

Amino Acid Sequence↗
PubMed1985

[Biochemical and nutritional value of seeds of the lima bean (Phaseolus lunatus) in comparison with those of the common bean (Phaseolus vulgaris)].

Composition in nutrients and antinutritional factors, digestibility and growth in rats fed diets prepared with raw and cooked beans of Ph. vulgaris, cultivar "Tacarigua", and Ph. lunatus cultivar "Tapiramo", are compared. Grains from both cultivars are similar in appearance, taste, nutritional value, and acceptability. Protein and phosphorus contents were greater in Ph. vulgaris than in Ph. lunatus seeds. The chemical score and availability of lysine were better in Ph. lunatus. Diets prepared with raw beans from Ph. lunatus resulted non toxic for the rats during a 12-day period of feeding. All rats fed with raw beans from Ph. Vulgaris died in the same period of time. Protein efficiency was better with cooked beans of Ph. lunatus. The cultivar "Tapiramo" (Ph. lunatus) is recommended for autoconsumption by small farmers.

Animals↗
PubMed2001

Effect of phosphorus availability on basal root shallowness in common bean.

Root gravitropism may be an important element of plant response to phosphorus availability because it determines root foraging in fertile topsoil horizons, and thereby phosphorus acquisition. In this study we seek to test this hypothesis in both two dimensional paper growth pouch and three-dimensional solid media of sand and soil cultures. Five common bean ( Phaseolus vulgaris L.) genotypes with contrasting adaptation to low phosphorus availability were evaluated in growth pouches over 6 days of growth, and in sand culture and soil culture over 4 weeks of growth. In all three media, phosphorus availability regulated the gravitropic response of basal roots in a genotype-dependent manner. In pouches, sand, and soil, the phosphorus-inefficient genotype DOR 364 had deeper roots with phosphorus stress, whereas the phosphorus-efficient genotype G19833 responded to phosphorus stress by producing shallower roots. Genotypes were most responsive to phosphorus stress in sand culture, where relative root allocation to the 0-3- and 3-6-cm horizons increased 50% with phosphorus stress, and varied 300% (3-6 cm) to 500% (0-3 cm) among genotypes. Our results indicate that (1) phosphorus availability regulates root gravitropic growth in both paper and solid media, (2) responses observed in young seedlings continue throughout vegetative growth, (3) the response of root gravitropism to phosphorus availability varies among genotypes, and (4) genotypic adaptation to low phosphorus availability is correlated with the ability to allocate roots to shallow soil horizons under phosphorus stress.

Biomass↗
PubMed2005

Resistance conferred against bean common mosaic virus by the incompletely dominant I locus of Phaseolus vulgaris is active at the single cell level.

A protoplast transfection system was used in Phaseolus vulgaris to study the incompletely dominant resistance locus I. The genetic materials in the study were cultivar 'Black Turtle Soup' lines nearly isogenic for I and their F1. Accumulation of bean common mosaic virus (BCMV; genus Potyvirus) RNA and virions was assayed following BCMV RNA electrotransfection of protoplasts from each genotype. BCMV RNA and virions accumulated in all genotypes tested but the relative rates of RNA accumulation differed. This suggests that the I allele is active at the single cell level and in a dosage-dependent fashion and supports previous work in this area.

Cells, Cultured↗
PubMed2004

Differential effects of cooked common bean (Phaseolus vulgaris) and lentil (Lens esculenta puyensis) feeding on protein and nucleic acid contents in intestines, liver and muscles in rats.

AIM: Our aim was to investigate the influence of legume feeding on the protein and nucleic acid content of intestinal tissues and muscles. METHODS: Growing male Wistar rats were fed ad libitum on balanced diets containing cooked common bean or lentil as the unique protein source (180 g.kg(-1) dry matter) for 20 days. The control group was pair-fed with an iso-energetic, iso-nitrogenous balanced casein diet. RESULTS: Intestinal tissues were heavier in the legume-fed groups and higher relative mass (g per 100 g body mass) of protein, RNA and DNA were found in these tissues as compared to those of the control groups. In liver protein and RNA masses were significantly lower in the bean group than in the control group but the DNA content was not different in the legume and in the control groups. In gastrocnemius and soleus muscles, there was no significant effect of legume feeding on the fresh mass and on the protein and RNA contents, but the DNA content of the m. gastrocnemius was significantly lower in both legume groups than in their control group. The ribosomal capacity of intestines, liver and muscles was not significantly affected by legume feeding. CONCLUSION: Legume feeding had a trophic effect on both proximal and distal intestinal tissues; dietary fiber appears to be the main cause of this effect.

Animals↗
PubMed2001

Tracing evolutionary and developmental implications of mitochondrial stoichiometric shifting in the common bean.

The recombination and copy number shifting activities of the plant mitochondrial genome are widely documented across plant genera, but these genome processes have not been as well examined with regard to their roles in plant evolution. Because of the extensive plant collections of Phaseolus spp and the degree to which cytoplasmic male sterility (cms) has been characterized in the common bean, this system would be valuable for investigating mitochondrial genome dynamics in natural populations. We have used the cms-associated sequence pvs-orf239 as a mitochondrial genetic marker for these studies and have demonstrated its universal presence throughout a diversity of undomesticated Phaseolus lines. Within these populations, the pvs-orf239 sequence is present in high copy number in approximately 10% of the lines, but substoichiometric in all others. This mitochondrial sequence, derived apparently by at least two recombination events, is well conserved with two point mutations identified that are both apparently silent with regard to the sterility phenotype. A putative progenitor sequence was identified in Phaseolus glabelus in substoichiometric levels, suggesting that the present-day pvs-orf239 sequence was likely introduced substoichiometrically. Copy number shifting within the mitochondrial genome results in a 1000- to 2000-fold change, so that substoichiometric forms are estimated at less than one copy per every 100 cells. On the basis of PCR analysis of root tips, we postulate that a mitochondrial "transmitted form" resides within the meristem to assure transmission of a complete genetic complement to progeny.

Cloning, Molecular↗
PubMed2003

Predicting progeny performance in common bean (Phaseolus vulgaris L.) using molecular marker-based cluster analysis.

Recovery of superior individuals from a cross based solely on the phenotypic characteristics ofsingle-plant selections is inefficient because some traits, like yield, have low heritabilities, or because it is difficult to create the correct conditions for selection, as with disease resistance. In contrast, molecular markers are highly heritable and unaffected by environmental conditions. The objective of this study was to investigate the potential of molecular markers to identify superior lines in a breeding population by examining relationships between genetic distances (GDs) and phenotypic data for eight agronomic and architectural traits (branch angle, height, hypocotyl diameter, lodging, maturity, upper pods, pods per plant, and yield) obtained from three locations over a two-year period. From an elite common bean (Phaseolus vulgaris L.) cross, 110 recombinant inbred lines (RILs) and the two parents were screened with 116 random amplified polymorphic DNA (RAPD) markers. Pairwise GD values were calculated between each line and a selected "target" (the parent 'OAC Speedvale') using the Jaccard method and correlated to the trait data. The correlations were low and non-significant for all traits, except for branch angle (r = 0.30), maturity (r = -0.25), and pods per plant (r = 0.35). The lines were also grouped according to their cluster-based GD from the target parent using UPGMA cluster analysis. Trait data of lines within groups were combined and correlated to cluster-based GD. Correlation values were large and significant for all traits. Additionally, one-half of the top 10 yielding lines and nearly one-third of the best phenotypically ranked lines were present within the 13% of lines clustered nearest the target. A selection method using marker-based cluster analysis (MBCA) is suggested to assist phenotypic selection by directing a breeder's attention to a subsample of the population containing a high proportion of superior lines.

Cluster Analysis↗
PubMed1988

Culinary practices and consumption characteristics of common beans at the rural home level.

A survey was carried out on housewives of rural bean-producing areas for the purpose of collecting information on bean selection procedures, culinary practices and bean consumption in rural homes from four departments of northeastern Guatemala. The study sample consisted of 10 families selected at random from each of 19 municipalities. The questionnaire used had been pretested in two rural communities not included in the study sample. As already shown by other INCAP surveys of this nature, it was found that all families consumed common black beans, and on rare occasions beans of different color or species. With respect to cooking practices, it was found that beans are not usually soaked prior to cooking, and that few housewives add salt at the beginning of cooking. Two fractions are usually obtained, the cooked beans and the cooking broth. Another finding was that beans are consumed in four preparations: cooked whole, strained and fried, ground and fried, and cooked and fried. Bean broth is consumed by both adults and children, and a thick broth is preferred since it is considered to be more nutritious and better tasting. Children begin to consume bean broth as early as two months of age, and beans at 14 months. The average amount of broth ingested was 62 g, and whole cooked beans, 70 g. Mothers surveyed indicated that broth was administered to children because: it was nutritious (56% of the mothers); in order to teach children how to eat (15%); or because of its taste (9%). Regarding the hard-to-cook phenomenon, 32% of the housewives indicated that storage time was responsible for it, while 17% attributed this condition to bad seeds, and 18% to sun-drying or inadequate postharvest practices and to their preparation for consumption. Although the survey confirmed the importance that common beans have in rural diets, the main finding was that the first bean food product fed to children is the bean-cooking liquor or broth, which on the average contains 8.52% of total solids and 1.48% protein.

Cooking↗
PubMed2003

Rhizobium etli and Rhizobium gallicum nodulate common bean (Phaseolus vulgaris) in a traditionally managed milpa plot in Mexico: population genetics and biogeographic implications.

The stability of the genetic structure of rhizobial populations nodulating Phaseolus vulgaris cultivated in a traditionally managed milpa plot in Mexico was studied over three consecutive years. The set of molecular markers analyzed (including partial rrs, glnII, nifH, and nodB sequences), along with host range experiments, placed the isolates examined in Rhizobium etli bv. phaseoli and Rhizobium gallicum bv. gallicum. Cluster analysis of multilocus enzyme electrophoresis and plasmid profile data separated the two species and identified numerically dominant clones within each of them. Population genetic analyses showed that there was high genetic differentiation between the two species and that there was low intrapopulation differentiation of the species over the 3 years. The results of linkage disequilibrium analyses are consistent with an epidemic genetic structure for both species, with frequent genetic exchange taking place within conspecific populations but not between the R. etli and R. gallicum populations. A subsample of isolates was selected and used for 16S ribosomal DNA PCR-restriction fragment length polymorphism analysis, nifH copy number determination, and host range experiments. Plasmid profiles and nifH hybridization patterns also revealed the occurrence of lateral plasmid transfer among distinct multilocus genotypes within species but not between species. Both species were recovered from nodules of the same plants, indicating that mechanisms other than host, spatial, or temporal isolation may account for the genetic barrier between the species. The biogeographic implications of finding an R. gallicum bv. gallicum population nodulating common bean in America are discussed.

Agriculture↗