Pinto Villa Research Articles

<b>Mechanisms for water-use efficiency between bean cultivars tolerant to drought are different

The water-use efficiency (WUE) has been proposed as an alternative to mitigate the effects of climate change in agriculture and to reduce pressure on water resources, mainly in species with high water requirements which are grown under rainfed conditions. The aim of this study was to characterize the instantaneous, integral and molecular WUE of four bean cultivars contrasting in their response under limiting water conditions to compare the component mechanisms of this trait between drought tolerant and susceptible cultivars. Results indicated that tolerant cultivars increased their instantaneous WUE in comparison to susceptible ones; however, there was a difference between cultivars since Pinto Villa had a higher stomatal conductance and transpiration rate, leading to a higher water cost to produce seed than Pinto Saltillo. Furthermore, ycf2, rrn16, rpoC2, hardy, ndhK, erecta and ycf1 WUE genes were only overexpressed in Pinto Saltillo under limited water conditions, which turned out to be the most WUE efficient cultivar. Therefore, the component mechanisms of WUE are different even between drought tolerant cultivars and the mechanisms by which the tolerant cultivars increased their instantaneous and integral WUE were different.

Rendimiento de frijol común (Phaseolus vulgaris L.) y Tépari (Phaseolus acutifolius A. Gray) bajo el método riego-sequia en Chihuahua


 
 
 La sequía es el factor que más limita la producción de frijol de temporal (Phaseolus vulgaris L.) en México. El objetivo del presente trabajo fue identificar frijol común y Tépari más resistentes a la sequía que Pinto Villa y Pinto Saltillo. Los experimentos en 2010 y 2011 se establecieron en la Estación Experimental de INIFAP en Bachiniva, Chihuahua. Se evaluaron siete variedades de frijol de diferente raza, hábito de crecimiento y contrastantes en la respuesta a la sequía. Se utilizó el método riego-sequía con dos riegos de auxilio durante el ciclo más la precipitación en el año (351 mm) y el tratamiento de sequía solo con la precipitación en el año. Se evaluó días a floración, días a madurez fisiológica y el rendimiento de semilla en kg ha-1. Se utilizó un diseño experimental en bloques completos al azar con cuatro repeticiones. Cada repetición consto de dos surcos de 5 m en 2010 y un surco de 5 m por repetición en 2011. Se utilizó la fórmula de fertilización 30-50-00. En 2010 el genotipo más estable fue Rosa La Bufa con tan sólo 10.5% de incremento del rendimiento en riego con respecto a sequía. En 2011los genotipos más estables fueron tépari café con 152.5% y Rosa La Bufa con 198.4%.
 
 

Contenido de osmoprotectores, ácido ascórbico y ascorbato peroxidasa en hojas de frijol sometidas a estrés por sequía


 
 
 La acumulación de osmoprotectores en plantas que se encuentran en condiciones de estrés por sequía, le permiten contrarrestar el efecto negativo que le provoca dichas condición ambiental. En el presente trabajo se estudió la respuesta bioquímica de plantas de frijol con base en la acumulación de carbohidratos solubles, almidón, ácido ascórbico, así como la actividad de la enzima ascorbato peroxidasa, en las hojas simples y en las dos primeras hojas trifolioladas de la variedad resistente, Pinto Villa, y la susceptible a sequía Bayo Madero, ambas de raza Durango. Las plantas de ambas variedades se sometieron a tres tratamientos de humedad, riego, sequía y riego de recuperación. Con respecto a Bayo Madero, Pinto Villa mostró una mayor respuesta de tolerancia al estrés de humedad relacionada con: altas concentraciones de prolina, de ácido ascórbico y mayor actividad de la enzima ascorbato peroxidasa, ésta última en el tratamiento de sequía. En relación a la cantidad de carbohidratos solubles, Bayo Madero fue estadísticamente superior (p< 0.05) a Pinto Villa, lo cual puede indicar que estos compuestos están siendo utilizados por la planta para sintetizar otros compuestos osmoprotectores como la prolina y que pueden considerarse como indicadores importantes para clasificar especies vegetales que sobrevivan y cumplan su ciclo biológico bajo condiciones de estrés de humedad.
 
 

Identification of a set of genes from genotypes of common bean tolerant and susceptible to water stress for a macroarray-based selection strategy

Globally, drought is the main factor that reduces common bean yield. For this reason, breeding alternatives, such as molecular marker-assisted selection, that focus on various functional genes directly involved in the response to water stress, such as those encoding late embryogenesis abundant (LEA), early response to dehydration (ERD), and dehydrin proteins, have been implemented. The aim of this study was to identify differentially expressed genes of Phaseolus vulgaris in drought-tolerant cultivars Pinto Saltillo (PS) and Pinto Villa (PV), and drought-susceptible cultivars Bayo Madero (BM) and Canario 60 (C60) in vegetative and reproductive stages. Relative water content (RWC) in leaf tissue was measured. Twenty-eight P. vulgaris genes obtained from GenBank and from a subtractive suppressive library from the PS cultivar were analysed, and their expression profiles were examined by reverse transcription polymerase chain reaction (RT-PCR). Then, cDNA arrays were developed and hybridised to confirm expression which was finally validated by quantitative PCR (qPCR). The usefulness of the identified genes as selection criteria for the tolerance of different genotypes to drought was examined using cDNA arrays. Expression of 21 genes was induced by drought. The cDNA arrays confirmed that expression of 19 of these genes increased in the vegetative stage upon exposure to the drought, and a higher expression was observed in the reproductive stage compared with vegetative stage V4. Only five genes induced by the drought were found to have a lower expression in the susceptible cultivars compared with the tolerant ones. During recovery after the drought in the reproductive stage, 13 of the 21 induced genes remained transcriptionally active including LEA3 and dehydrin. The RWC during the drought in vegetative stage V4 decreased by about 55 % in all cultivars, but at the onset of flowering, it increased to 80 % in PV and PS. In contrast, in the susceptible cultivars, it remained at 55 %. Using qPCR validation, expression induction was confirmed in the drought-tolerant cultivars. Polyubiquitin2, LEA3, LEA4, and dehydrin were useful genes for selecting drought-tolerant genotypes under field conditions.

Identification of novel drought-tolerant-associated SNPs in common bean (Phaseolus vulgaris).

Common bean (Phaseolus vulgaris L.) is a leguminous in high demand for human nutrition and a very important agricultural product. Production of common bean is constrained by environmental stresses such as drought. Although conventional plant selection has been used to increase production yield and stress tolerance, drought tolerance selection based on phenotype is complicated by associated physiological, anatomical, cellular, biochemical, and molecular changes. These changes are modulated by differential gene expression. A common method to identify genes associated with phenotypes of interest is the characterization of Single Nucleotide Polymorphims (SNPs) to link them to specific functions. In this work, we selected two drought-tolerant parental lines from Mesoamerica, Pinto Villa, and Pinto Saltillo. The parental lines were used to generate a population of 282 families (F3:5) and characterized by 169 SNPs. We associated the segregation of the molecular markers in our population with phenotypes including flowering time, physiological maturity, reproductive period, plant, seed and total biomass, reuse index, seed yield, weight of 100 seeds, and harvest index in three cultivation cycles. We observed 83 SNPs with significant association (p < 0.0003 after Bonferroni correction) with our quantified phenotypes. Phenotypes most associated were days to flowering and seed biomass with 58 and 44 associated SNPs, respectively. Thirty-seven out of the 83 SNPs were annotated to a gene with a potential function related to drought tolerance or relevant molecular/biochemical functions. Some SNPs such as SNP28 and SNP128 are related to starch biosynthesis, a common osmotic protector; and SNP18 is related to proline biosynthesis, another well-known osmotic protector.

Mejoramiento de la resistencia a la sequía del frijol común en México.

Since its inception in 1980, the breeding project for adaptation to drought has been based on crop phenology (escape or matching between biological cycle and water availability), and seed yield in multilocational trials under water stress conditions. For yield evaluation, bean genotypes were grouped on the basis of similar growth habits and phenology. Selection of segregating families according to adaptation to drought was postponed up until the F4 generation or subsequent ones, using incomplete block designs. Selection in early generations was based on moderate to high heretability traits such as biological cycle, grain phenology (color, size, shine), and reaction to diseases. Eight improved Durango bean varieties that are tolerant to temporary water deficits (Pinto Villa, Pinto Mestizo, Pinto Bayacora, Negro Altiplano, Negro Sahuatoba, Negro Durango, Bayo Victoria, and Azufrado Namiquipa) have been given to bean producers from Mexico’s semi-arid highlands. In 1998 seeds of two new varieties (Pinto Zapata and Flor de Mayo 2000) will be increased and registered. Their main traits are: type III indeterminate growth habits, short biological cycle, fast transition from vegetative to reproductive phase, and high yield rates.

Reacción de variedades de frijol común a Fusarium spp. y Rhizoctonia solani en el altiplano de México.

Two trials were carried out at Texcoco, State of Mexico: in the first one 32 bean genotypes were sowed under irrigation on April 24 and May 8, 1997, in order to study their reaction to Fusarium spp. in a naturally infested soil. In the second one, protection against Fusarium spp. y R. solani was evaluated by applying captan or benomyl to the seeds of eight genotypes. In the first trial most genotypes were susceptible to Fusarium spp., particularly in the third evaluation (R6), on both sowing dates; with the exception of Negro Tacana and Amarillo de Calpan, both of which showed an intermediate reaction in the second sowing date. Incidence of Fusarium spp. was higher than 80%. On the first sowing date the Pinto Villa and Negro Perla genotypes were moderately resistant. In the second experiment the chemical treatment produced more vigorous seedlings. Pinto Villa and BAT 477 genotypes showed lower severity scores when treated with benomyl and captan, respectively. It was observed that plants of resistant genotypes survived because they developed adventitious roots.

Effect of infrared heating on the physicochemical properties of common bean (Phaseolus vulgaris L.) flour

The objective of the presented work was to determine the effect of infrared heating on the physicochemical characteristics of Mexican common bean flour. Variety of Pinto Villa beans were used in the present experiment. Infrared heating was fitted at 0, 10, 20 and 40 Hz. Power absorption, relative crystallinity by X-ray diffraction, proximate analysis, water absorption capacity, water absorption index, viscosity, and total, resistant and available starch methods were used to characterize the common bean flour. X-ray diffraction data showed changes in the diffraction patterns attributable to the possible formation of amylose–lipid complex. Water absorption capacity (WAC) and water absorption index (WAI), as well viscosity profiles, were affected by changes in the starch and protein content. Sample with highest power absorbed (20 Hz) show highest values of WAC and WAI and lowest value of viscosity. The content of resistant starch (RS) was highest at medium absorbed power (10 Hz).

Differential Expression of Genes for Tolerance to Salt Stress in Common Bean (Phaseolus vulgaris L.)

Crop production is severely reduced by abiotic stresses such as drought, salinity, and extreme temperatures. The common bean (Phaseolus vulgaris L.) is the most important legume for direct human consumption worldwide. In order to identify genes that are induced during the salt stress response in a tolerant bean cultivar (Pinto Villa), suppression subtractive hybridization was performed. The tester cDNA was prepared from the leaves of bean plants treated with 200 mM NaCl for 2 and 5 days and the driver cDNA from the leaves of the unstressed plants. After hybridizing and cloning, forward subtraction cDNA library was obtained, and 67 unigenes were generated from 380 randomly picked clones. These unigenes were classified into ten categories according to their putative function assigned by the Gene Ontology database. Six unigenes were selected to analyze for differential expression by qRT-PCR, including heat shock proteins such as PvDnaJ3 and PvHSP90, late embryogenesis abundant proteins such as PvLEA-18 and dehydrin PvERD10, the transcription factor PvATHB-7, and a protein of unknown function that contains a short glycine-rich domain (PvGRDP1). These genes were found differentially expressed in a time-dependent manner in bean tissues under salt stress. Data generated from this study suggests that the response of the Pinto Villa bean cultivar to salt stress could be through the regulation of genes such as chaperones, transcription factors, and late embryogenesis abundant proteins which are involved in plant tolerance.

The relations between antioxidant enzymes and chlorophyll fluorescence parameters in common bean cultivars differing in sensitivity to drought stress

Effects of the antioxidant system and chlorophyll fluorescence on drought tolerance of four common bean (Phaseolus vulgaris L.) cultivars were studied. The cultivars were positioned in the order of a decrease in their drought tolerance: Yakutiye, Pinto Villa, Ozayse, and Zulbiye on the basis of changes in the water potential, stomatal conductance, photosynthetic pigment content, and lipid peroxidation. Under drought conditions, the level of H2O2 was not changed in cv. Pinto Villa but decreased in other cultivars. Antioxidant enzymes (superothide dismutase (SOD), guaiacol peroxidase (GPX), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR)) were generally activated in all cultivars. Interestingly, CAT, APX, and GR activities were not changed in cv. Pinto Villa, APX activity decreased in cv. Yakutiye, and CAT activity was not changed in cv. Zulbiye. The increases in SOD and GPX activities in cv. Ozayse were higher than in other cultivars. Drought stress reduced the effective quantum yield of PS2 (ΦPS2) and the photochemical quenching (qp), while it increased nonphotochemical quenching (NPQ) in all cultivars. The reduction or increase was more pronounced in cv. Zulbiye. There were generally significant correlations between qp, NPQ, and ROS scavenging by SOD and APX. Also, there were significant correlations between SOD and qp in tolerant cultivars and APX and qp in sensitive ones. The results indicate that activation of SOD and APX was closely related to the efficiency of PS2 in common bean cultivars. This interaction was essential for protection of photosystems and plant survival under drought.

Effect of co-inoculation with mycorrhiza and rhizobia on the nodule trehalose content of different bean genotypes.

Studies on Rhizobium-legume symbiosis show that trehalose content in nodules under drought stress correlates positively with an increase in plant tolerance to this stress. Fewer reports describe trehalose accumulation in mycorrhiza where, in contrast with rhizobia, there is no flux of carbohydrates from the microsymbiont to the plant. However, the trehalose dynamics in the Mycorrhiza-Rhizobium-Legume tripartite symbiosis is unknown. The present study explores the role of this tripartite symbiosis in the trehalose content of nodules grown under contrasting moisture conditions. Three wild genotypes (P. filiformis, P. acutifolis and P. vulgaris) and two commercial genotypes of Phaseolus vulgaris (Pinto villa and Flor de Mayo) were used. Co-inoculation treatments were conducted with Glomus intraradices and a mixture of seven native rhizobial strains, and trehalose content was determined by GC/MS. The results showed a negative effect of mycorrhizal inoculation on nodule development, as mycorrhized plants showed fewer nodules and lower nodule dry weight compared to plants inoculated only with Rhizobium. Mycorrhizal colonization was also higher in plants inoculated only with Glomus as compared to plants co-inoculated with both microsymbionts. In regard to trehalose, co-inoculation negatively affects its accumulation in the nodules of each genotype tested. However, the correlation analysis showed a significantly positive correlation between mycorrhizal colonization and nodule trehalose content.

Effect of Co-Inoculation with Mycorrhiza and Rhizobia on the Nodule Trehalose Content of Different Bean Genotypes

Studies on Rhizobium-legume symbiosis show that trehalose content in nodules under drought stress correlates positively with an increase in plant tolerance to this stress. Fewer reports describe trehalose accumulation in mycorrhiza where, in contrast with rhizobia, there is no flux of carbohydrates from the microsymbiont to the plant. However, the trehalose dynamics in the Mycorrhiza-Rhizobium-Legume tripartite symbiosis is unknown. The present study explores the role of this tripartite symbiosis in the trehalose content of nodules grown under contrasting moisture conditions. Three wild genotypes (P. filiformis, P. acutifolis and P. vulgaris) and two commercial genotypes of Phaseolus vulgaris (Pinto villa and Flor de Mayo) were used. Co-inoculation treatments were conducted with Glomus intraradices and a mixture of seven native rhizobial strains, and trehalose content was determined by GC/MS. The results showed a negative effect of mycorrhizal inoculation on nodule development, as mycorrhized plants showed fewer nodules and lower nodule dry weight compared to plants inoculated only with Rhizobium. Mycorrhizal colonization was also higher in plants inoculated only with Glomus as compared to plants co-inoculated with both microsymbionts. In regard to trehalose, co-inoculation negatively affects its accumulation in the nodules of each genotype tested. However, the correlation analysis showed a significantly positive correlation between mycorrhizal colonization and nodule trehalose content.

Relationship between carbohydrate partitioning and drought resistance in common bean

Drought is a major yield constraint in common bean (Phaseolus vulgaris L.). Pulse-chase (14)C-labelling experiments were performed using Pinto Villa (drought resistant) and Canario 60 (drought sensitive) cultivars, grown under optimal irrigation and water-deficit conditions. Starch and the radioactive label incorporated into starch were measured in leaves and pods at different time points, between the initiation of pod development and the production of mature pods. The water-stress treatment induced a higher starch accumulation in the drought-resistant cultivar pods than in those of the drought-sensitive cultivar. This effect was more noticeable during the early stages of pod development. Consistently, a reduction of starch content occurred in the leaves of the drought-resistant cultivar during the grain-filling stage. Furthermore, a synchronized accumulation of sucrose was observed in immature pods of this cultivar. These data indicate that carbohydrate partitioning is affected by drought in common bean, and that the modulation of this partitioning towards seed filling has been a successful strategy in the development of drought-resistant cultivars. In addition, our results suggest that, in the drought-resistant cultivar, the efficient carbon mobilization towards the seeds in response to water limitation is favoured by a mechanism that implies a more effective sucrose transport.

Systemic nature of drought-tolerance in common bean

The response to drought at the physiological and molecular levels was studied in two common bean varieties with contrasting susceptibility to drought stress. A number of genes were found to be up-regulated in the tolerant variety Pinto Villa relative to the susceptible cultivar, Carioca. The products of these genes fell in different functional categories. Further analyses of selected genes, consisting of their spatial differential expression and in situ mRNA accumulation patterns displayed interesting profiles. The drought-tolerant variety displayed a more developed root vasculature in drought conditions, when compared to the susceptible tropical bean Carioca. The in situ localization of three selected genes indicated the accumulation of their corresponding mRNAs in companion cells, sieve tubes and in developing phloem, suggesting that these, and/or the encoded proteins could constitute phloem-mobile signals. Indeed, a number of transcripts that are induced in response to water deficit accumulate in the phloem in other plant species, suggesting a general phenomenon. Moreover, the analysis of drought stress in plant varieties with contrasting tolerance to such stimulus will help to determine the role of differential expression of specific genes in response to such phenomenon, as well as other biochemical, morphological and physiological features in both cultivars.

Physical Properties of Extruded Products from Three Mexican Common Beans (Phaseolus vulgaris L.) Cultivars

The physical properties of extruded products from three Mexican common bean cultivars were investigated. Common bean cultivars Flor de Mayo, Pinto Villa and Bayo Victoria from the same harvesting season (2006) were used in this work. Beans were milled and the flour was hydrated to 24, 26 and 28 g of water/100 g of dry weight. Two temperatures, 130 degrees C and 165 degrees C at the end of the extruder barrel without die, were experimented. Common bean flour extrudates were evaluated for water absorption index (WAI), water absorption capacity (WAC), oil absorption capacity (OAC), and emulsifying capacity (EC). Flor de Mayo extrudates showed the highest WAC and WAI values. Thus starch from Flor de Mayo beans showed minor restricted water availability. In all cases, the OAC of extruded products was lower than the crude bean flour. The EC for Bayo Victoria flour increased as a consequence of the extrusion process. The EC for Flor de Mayo was higher at lower temperature and lower moisture content than Pinto Villa and Bayo Victoria beans. EC behavior of Pinto Villa was similar to Bayo cultivar. These results indicate that it is possible to produce new extruded products with good physical properties from these common bean cultivars.

Effect of bean size on the physicochemical properties of different bean varieties (Phaseolus vulgaris)

The textural properties and hydration capacity of legumes in general and of beans in particular is a subject about which much research has been done. However, at present there is still no practical answer to the “Hard to Shell” and “Hard to Cook” effects. The present investigation studies 14 varieties of recently picked beans that were grown in Mexico. The beans were classified by size using K-means multivariate analysis, and three fractions were obtained for each variety: large (L), medium (M) and small (S). These fractions were evaluated for hardness, water absorption rate and total water absorption. It was found that the small bean fraction of the varieties studied had the lowest hardness values in comparison to the medium and large fractions. This same behaviour was observed for the water absorption rate and total water absorption; the smaller beans absorbed water more quickly and had greater total water absorption than the other fractions. One kind of bean (Pinto Villa) proved to be much harder than ...

Differential accumulation of mRNAs in drought‐tolerant and susceptible common bean cultivars in response to water deficit

The physiological response to drought was measured in two common bean varieties with contrastive susceptibility to drought stress. A subtractive cDNA library was constructed from the two cultivars, Phaseolus vulgaris'Pinto Villa' (tolerant) and 'Carioca' (susceptible). 18 cDNAs displayed protein-coding genes associated with drought, cold and oxidative stress, signal transduction, plant defense, chloroplast function and unknown function. A cDNA coding for an aquaporin (AQP) was selected for further analyses. The open reading frames (ORFs) of AQPs from 'Pinto Villa' and 'Carioca' were compared and despite their similarity, accumulated differentially in the plant organs, as demonstrated by Northern blot and in situ hybridization. A phylogenetic analysis of the deduced amino acid sequence with other AQPs suggested a tonoplast-located protein. Under drought conditions, the levels of AQP mRNA from the susceptible cultivar decreased to undetectable levels; by contrast, 'Pinto Villa' mRNA was present and restricted the phloem tissue. This would allow 'Pinto Villa' to maintain vascular tissue functions under drought stress.

Isolation of dehydration-responsive genes in a drought tolerant common bean cultivar and expression of a group 3 late embryogenesis abundant mRNA in tolerant and susceptible bean cultivars.

Drought is one of the main constraints for common bean (Phaseolus vulgaris L.) production in Latin America. The aim of this work was to identify upregulated genes in the drought-tolerant common bean cv. Pinto Villa, grown under water-deficit conditions. Twenty-eight cDNAs representing differentially-expressed mRNAs in roots and/or leaves were isolated via suppression subtractive hybridisation. Their expression profiles in plants under intermediate and severe dehydration stress were tested. Three cDNAs corresponded to genes already described as associated to drought stress in P. vulgaris, 12 were known P. vulgaris sequences without previous association with drought response, and 13 were new P. vulgaris sequences. Analysis of the deduced proteins encoded by the cDNAs revealed putative functions in cellular protection, sugar metabolism, and protein synthesis, folding and turnover. Additionally, a new member of group 3 late embryogenesis abundant (LEA) genes (PvLEA3) was cloned and its complete sequence was obtained. Given the lack of reports comparing expression of dehydration-responsive genes in bean cultivars with different response to drought, the expression of PvLEA3 transcript in five bean cultivars from different origin was analysed. The induction of PvLEA3 was directly associated with the level of drought tolerance in the cultivars studied.

Proline accumulation in two bean cultivars under salt stress and the effect of polyamines and ornithine

Proline accumulation in two different bean (Phaseolus vulgaris L.) cultivars, one drought-sensitive (Canario 60) and one drought-resistant (Pinto Villa) was investigated. Both tolerated salt concentrations up to 150 mM NaCl, but the sensitive Canario 60 did not survive at 400 mM NaCl. In response to salt stress, both cvs. accumulated proline in all the analyzed tissues, the lowest contents were detected in roots. Pinto Villa accumulated higher proline concentrations than Canario 60 only at 400 mM NaCl. The addition of polyamines or ornithine increased proline content in plant tissues without stress, while they decreased it under salt stress.

Effect of pressure cooking on the antioxidant activity of extracts from three common bean ( Phaseolus vulgaris L.) cultivars

Common bean is a traditional legume that is becoming more attractive because of its dietary and associated health benefits. Flor de Mayo M38, Pinto Villa and Bayo Victoria, three common bean cultivars developed in Mexico, were tested for phenol content and free radical scavenging activity before and after autoclaving. Independent analysis of seed coat and cotyledon was performed for each cultivar. Longer cooking times enhanced diffusion of phenols from seed coats to cooking water and from there to cotyledons. Cooking waters showed a remarkable activity similar to crude seed coats extracts although their phenol content was 80% lower.