Phytochemical Profile and Composition of Chickpea (Cicer arietinum L.): Varietal Differences and Effect of Germination under Elicited Conditions.

Germination is a simple process that improves the nutritional and medicinal values of seeds such as chickpeas. However, the detailed analysis of the phytochemical profile after chemical elicitation during chickpea germination is indispensable when making inferences about its biological properties. Therefore, an evaluation was made of the effect of the chemical inducers salicylic acid (SA, 1 and 2 mM), chitosan (CH, 3.3 and 7 µM), and hydrogen peroxide (H2O2, 20 and 30 mM) during germination at 25 °C with 70% RH for 4 days on the content of antinutritional and bioactive compounds, including phenolics, sterols, and saponins, in three Mexican chickpea varieties (Blanoro, Patron, and San Antonio) using UPLC-ELSD-ESI-QqQ-MS/MS, UPLC-DAD-ESI-QqQ-MS/MS, and HPLC-DAD-sQ-MS. The highest increase in phenolics and saponins was found in the Blanoro sprouts induced with SA 2 mM, whereas the highest phytosterol content was detected in San Antonio sprouts induced with CH 7 µM. In addition, significant increases in mono-, di-, and oligosaccharides and decreases in antinutritional contents were achieved after germination with most of the elicitation conditions. More importantly, we identified new compounds in chickpea sprouts, such as the lignans matairesinol and secoisolariciresinol, the phenolic compounds epicatechin gallate and methyl gallate, some phytosterols, and the saponin phaseoside 1, which further increased after chemical elicitation.

Carbon-concentrating mechanisms in pods are key elements for terminal drought resistance in Phaseolus vulgaris.

Common bean (Phaseolus vulgaris L.) is one of the most consumed legumes in the human diet and a substantial source of dietary protein. A major problem for this rainfed crop is the decrease in grain yield caused by prolonged drought periods during the reproductive stage of plant development (terminal drought). Terminal drought remains a prevailing threat to the farming of this staple, with losses reaching >80%. Based on the high correlation between the resistance of common bean to terminal drought and efficient photoassimilate mobilization and biomass accumulation in seeds, we aimed to identify mechanisms implicated in its resistance to this stress. We used two representative Durango race common bean cultivars with contrasting yields under terminal drought, grown under well-watered or terminal drought conditions. Using comparative transcriptomic analysis focused on source leaves, pods, and seeds from both cultivars, we provide evidence indicating that under terminal drought the resistant cultivar promotes the build-up of transcripts involved in recycling carbon through photosynthesis, photorespiration, and CO2-concentrating mechanisms in pod walls, while in seeds, the induced transcripts participate in sink strength and respiration. Physiological data support this conclusion, implicating their relevance as key processes in the plant response to terminal drought.

Common Bean Seeds Obtained by Plant Water Restriction Ameliorates Obesity-Associated Cardiovascular Risk and Insulin Resistance.

The inclusion of beans in the diet has been recommended for obesity control. However, its beneficial effect varies depending on agroclimatic factors acting during plant development. The antiobesogenic capacity of Dalia bean (DB) seeds obtained by water restriction (WR) during the vegetative or reproductive stage of plant growth (50/100 and 100/50% of soil moisture in vegetative/reproductive stage, respectively), during the whole cycle (50/50), and well-watered plants (100/100) was researched. After phytochemical characterization, harvested beans from each experimental unit were pooled among treatments, based on a multivariate canonical discriminant analysis considering concentration of non-digestible carbohydrates (total, soluble and insoluble dietary fiber and resistant starch), phenolic compounds (total phenols, flavonoids, anthocyanins and condensed tannins) and total saponins, which showed no differences among replicas of each treatment. Obesity was induced in rats (UAZ-2015-36851) with a high fat diet (HFD) for four months. Afterwards, rats were fed with the HFD supplemented with 20% of cooked DB for three months. During treatment, 100/50 beans, improved blood triglycerides, cholesterol, and glucose, and alleviated early insulin resistance (IR) related to inhibition of lipase, α-amylase and -glucosidase activity. After sacrifice, a hypolipidemic capacity and atherogenic risk reduction was observed, especially from the 100/50 treatment, suggesting that intake of DB obtained from WR may prevent IR and dyslipidemia.

Diversity and Distribution of Viruses Infecting Wild and Domesticated Phaseolus spp. in the Mesoamerican Center of Domestication.

Viruses are an important disease source for beans. In order to evaluate the impact of virus disease on Phaseolus biodiversity, we determined the identity and distribution of viruses infecting wild and domesticated Phaseolus spp. in the Mesoamerican Center of Domestication (MCD) and the western state of Nayarit, Mexico. We used small RNA sequencing and assembly to identify complete or near-complete sequences of forty-seven genomes belonging to nine viral species of five genera, as well as partial sequences of two putative new endornaviruses and five badnavirus- and pararetrovirus-like sequences. The prevalence of viruses in domesticated beans was significantly higher than in wild beans (97% vs. 19%; p < 0.001), and all samples from domesticated beans were positive for at least one virus species. In contrast, no viruses were detected in 80-83% of the samples from wild beans. The Bean common mosaic virus and Bean common mosaic necrosis virus were the most prevalent viruses in wild and domesticated beans. Nevertheless, Cowpea mild mottle virus, transmitted by the whitefly Bemisia tabaci, has the potential to emerge as an important pathogen because it is both seed-borne and a non-persistently transmitted virus. Our results provide insights into the distribution of viruses in cultivated and wild Phaseolus spp. and will be useful for the identification of emerging viruses and the development of strategies for bean viral disease management in a center of diversity.

Steaming and Toasting Reduce the Nutrimental Quality, Total Phenols and Antioxidant Capacity of Fresh Kabuli Chickpea (Cicer arietinum L.).

Kabuli chickpea is traditionally consumed in Mexico. It is currently exported to countries including United States where its demand has recently increased. In order to demonstrate the effect of thermal processes on the quality of fresh chickpea, the objective of the present work was to evaluate some nutrimental and functional characteristics of raw, steamed and toasted chickpea. The partial chemical composition, total phenols, oligosaccharides, and antioxidant capacities were measured in five genotypes of chickpea. Steamed and toasted chickpea showed up to 8.4 and 25.8% less protein, respectively, than that of raw samples. Oligosaccharides, in general decreased in steamed and toasted fresh grain; however, verbascose increased on average 30.6 and 37.9% in steamed and toasted samples, respectively. Minor changes in total phenolic content were observed a result of the process. Trolox equivalent antioxidant capacity increased up to 3.5 times compared to that of antioxidant capacity of raw samples. Fresh chickpea grain, raw or processed, shows attractive nutritional and antioxidant properties that can contribute to the diet and health of the person who consumes it.

A new begomovirus isolated from a potyvirus-infected bean plant causes asymptomatic infections in bean and N. benthamiana.

In this work, a begomovirus isolated from a bean plant coinfected with the potyviruses bean common mosaic virus and bean common mosaic necrosis virus was characterized. The three viruses were detected by high-throughput sequencing and assembly of total small RNAs, but the begomovirus-related contigs did not allow precise identification. Molecular analysis based on standard DNA amplification techniques revealed the presence of a single bipartite virus, which is a novel begomovirus according to the current taxonomic criteria. Infectious clones were generated and agroinoculated into Phaseolus vulgaris and Nicotiana benthamiana plants. In all cases, viral DNA-A and DNA-B were detected in new growths, but no symptoms were observed, thus indicating that this virus produces asymptomatic infections in both host species.

Common bean seeds from plants subjected to severe drought, restricted- and full-irrigation regimes show differential phytochemical fingerprint.

Common bean is rich in phytochemicals like polyphenols, saponins, and steroidal glycosides, but its crop is greatly affected by drought. Thus, it is necessary to identify the irrigation regimes that improves its phytochemical profile without affecting seed yield. The aim of this study was to evaluate the differential effect of severe drought (SD), restricted irrigation (RI), and full irrigation (FI) on the phytochemical fingerprint of bean seeds through liquid chromatography-mass spectrometry and chemometric approach. Partial least square-discriminant analysis (PLS-DA) indicated that seeds from plants grown under RI where differentiated from those grown under FI and SD due to their high content of quercetin 3-O-rhamnoside and luteolin 7-O-glucoside. Regarding seed yield, no significant (p < 0.05) differences were observed between RI and FI, whereas SD decreased (p < 0.05) seed yield as compared to FI (47%). These results suggest that mild hydric stress (RI) can be used to induce phytochemicals without affecting seed yield.

Simultaneous Detection of Both RNA and DNA Viruses Infecting Dry Bean and Occurrence of Mixed Infections by BGYMV, BCMV and BCMNV in the Central-West Region of Mexico.

A multiplex reverse transcription polymerase chain reaction (RT-PCR) assay was developed to simultaneously detect bean common mosaic virus (BCMV), bean common mosaic necrotic virus (BCMNV), and bean golden yellow mosaic virus (BGYMV) from common bean leaves dried with silica gel using a single total nucleic acid extraction cetyl trimethyl ammonium bromide (CTAB) method. A mixture of five specific primers was used to amplify three distinct fragments corresponding to 272 bp from the AC1 gene of BGYMV as well as 469 bp and 746 bp from the CP gene of BCMV and BCMNV, respectively. The three viruses were detected in a single plant or in a bulk of five plants. The multiplex RT-PCR was successfully applied to detect these three viruses from 187 field samples collected from 23 municipalities from the states of Guanajuato, Nayarit and Jalisco, Mexico. Rates of single infections were 14/187 (7.5%), 41/187 (21.9%), and 35/187 (18.7%), for BGYMV, BCMV, and BCMNV, respectively; 29/187 (15.5%) samples were co-infected with two of these viruses and 10/187 (5.3%) with the three viruses. This multiplex RT-PCR assay is a simple, rapid, sensitive, and cost-effective method for detecting these viruses in the common bean and can be used for routine molecular diagnosis and epidemiological studies.

Effect of chemical stress on germination of cv Dalia bean (Phaseolus vularis L.) as an alternative to increase antioxidant and nutraceutical compounds in sprouts.

The aim of this study was to determine the effect of chitosan (CH), salicylic acid (SA) and hydrogen peroxide (H2O2) at different concentrations on the antinutritional and nutraceutical content, as well as the antioxidant capacity of bean sprouts (cv Dalia). All elicitors at medium and high concentrations reduced the antinutritional content of lectins (48%), trypsin inhibitor (57%), amylase inhibitor (49%) and phytic acid (56%). Sprouts treated with CH, SA and H2O2 (7µM; 1 and 2mM, and 30mM respectively) increased the content of phenolic compounds (1.8-fold), total flavonoids (3-fold), saponins (1.8-fold) and antioxidant capacity (37%). Furthermore, the UPLC-ESI-MS/MS analysis showed an increase of several nutraceutical compounds in bean sprouts treated with SA such as coumaric (8.5-fold), salicylic (115-fold), gallic (25-fold) and caffeic (1.7-fold) acids, as well as epigallocatechin (63-fold), rutin (41-fold) and quercetin (16.6-fold) flavonoids. The application of elicitors in bean seed during sprouting enhances their nutraceutical properties.

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.

Photosynthesis and chloroplast genes are involved in water-use efficiency in common bean.

A recent proposal to mitigate the effects of climatic change and reduce water consumption in agriculture is to develop cultivars with high water-use efficiency. The aims of this study were to characterize this trait as a differential response mechanism to water-limitation in two bean cultivars contrasting in their water stress tolerance, to isolate and identify gene fragments related to this response in a model cultivar, as well as to evaluate transcription levels of genes previously identified. Keeping CO2 assimilation through a high photosynthesis rate under limited conditions was the physiological response which allowed the cultivar model to maintain its growth and seed production with less water. Chloroplast genes stood out among identified genetic elements, which confirmed the importance of photosynthesis in such response. ndhK, rpoC2, rps19, rrn16, ycf1 and ycf2 genes were expressed only in response to limited water availability.

A non-digestible fraction of the common bean (Phaseolus vulgaris L.) induces cell cycle arrest and apoptosis during early carcinogenesis.

We have previously demonstrated that the non-digestible fraction (NDF) from common cooked beans (P. vulgaris L., cv Negro 8025) inhibits azoxymethane (AOM)-induced colon cancer and influences the expression of genes involved in the induction of apoptosis and cell cycle arrest through the action of butyrate. The objective of this study was to identify cell cycle alterations and morphological changes induced by treatment with AOM and to examine the formation of colonic aberrant crypt foci (ACF) in male Sprague Dawley rats fed with these beans. Rats were fed control diets upon arrival and were randomly placed into four groups after one week of acclimatization: control, NDF (intragastric administration), NDF + AOM and AOM. Rats treated with NDF + AOM exhibited a significantly lower number of total colonic ACF with a notable increase in the number of cells present in the G1 phase (83.14%); a decreased proliferation index was observed in the NDF + AOM group when compared to AOM group. NDF + AOM also displayed a higher number of apoptotic cells compared to AOM group. NDF of cooked common beans inhibited colon carcinogenesis at an early stage by inducing cell cycle arrest of colon cells and morphological changes linked to apoptosis, thus confirming previous results obtained with gene expression studies.

Phenotyping common beans for adaptation to drought.

Common beans (Phaseolus vulgaris L.) originated in the New World and are the grain legume of greatest production for direct human consumption. Common bean production is subject to frequent droughts in highland Mexico, in the Pacific coast of Central America, in northeast Brazil, and in eastern and southern Africa from Ethiopia to South Africa. This article reviews efforts to improve common bean for drought tolerance, referring to genetic diversity for drought response, the physiology of drought tolerance mechanisms, and breeding strategies. Different races of common bean respond differently to drought, with race Durango of highland Mexico being a major source of genes. Sister species of P. vulgaris likewise have unique traits, especially P. acutifolius which is well adapted to dryland conditions. Diverse sources of tolerance may have different mechanisms of plant response, implying the need for different methods of phenotyping to recognize the relevant traits. Practical considerations of field management are discussed including: trial planning; water management; and field preparation.

Physiological traits related to terminal drought resistance in common bean (Phaseolus vulgaris L.).

BACKGROUND: A major problem in common bean (Phaseolus vulgaris L.) agriculture is the low yield due to terminal drought. Because common beans are grown over a broad variety of environments, the study of drought-resistant genotypes might be useful to identify distinctive or common mechanisms needed for survival and seed production under drought. RESULTS: In this study the relationship between terminal drought resistance and some physiological parameters was analysed using cultivars contrasting in their drought response from two different gene pools. Trials were performed in three environments. As expected, drought treatments induced a decrease in leaf relative humidity and an increase in leaf temperature; however, when these parameters were compared between susceptible and resistant cultivars under optimal irrigation and drought, no significant differences were detected. Similar results were obtained for chlorophyll content. In contrast, analysis of relative water content (RWC) and stomatal conductance values showed reproducible significant differences between susceptible and resistant cultivars grown under optimal irrigation and drought across the different environments. CONCLUSIONS: The data indicate that drought-resistant cultivars maximise carbon uptake and limit water loss upon drought by increasing stomatal closure during the day and attaining a higher RWC during the night as compared with susceptible cultivars, suggesting a water balance fine control to achieve enough yield under drought.

Physiological analysis of common bean (Phaseolus vulgaris L.) cultivars uncovers characteristics related to terminal drought resistance.

Terminal drought is a major problem for common bean production because it occurs during the reproductive stage, importantly affecting seed yield. Diverse common bean cultivars with different drought susceptibility have been selected from different gene pools in several drought environments. To better understand the mechanisms associated with terminal drought resistance in a particular common bean race (Durango) and growth habit (type-III), we evaluated several metabolic and physiological parameters using two cultivars, Bayo Madero and Pinto Saltillo, with contrasting drought susceptibility. The common bean cultivars were submitted to moderate and severe terminal drought treatments under greenhouse conditions. We analyzed the following traits: relative growth rate, photosynthesis and transpiration rates, stomatal conductance, water-use efficiency, relative water content, proline accumulation, glycolate oxidase activity and their antioxidant response. Our results indicate that the competence of the drought-resistant cultivar (Pinto Saltillo) to maintain seed production upon terminal drought relies on an early response and fine-tuning of stomatal conductance, CO2 diffusion and fixation, and by an increased water use and avoidance of ROS accumulation.

Composition and chemopreventive effect of polysaccharides from common beans (Phaseolus vulgaris L.) on azoxymethane-induced colon cancer.

Common beans ( Phaseolus vulgaris L.) contain a high proportion of undigested carbohydrates (NDC) that can be fermented in the large intestine to produce short-chain fatty acids (SCFA) such as acetate, propionate, and butyrate. The objective of the present study was to evaluate the composition and chemopreventive effect of a polysaccharide extract (PE) from cooked common beans ( P. vulgaris L) cv. Negro 8025 on azoxymethane (AOM) induced colon cancer in rats. The PE induced SCFA production with the highest butyrate concentrated in the cecum zone: 6.7 +/- 0.06 mmol/g of sample for PE treatment and 5.29 +/- 0.24 mmol/g of sample for PE + AOM treatment. The number of aberrant crypt foci (ACF) and the transcriptional expression of bax and caspase-3 were increased, and rb expression was decreased. The data suggest that PE decreased ACF and had an influence on the expression of genes involved in colon cancer for the action of butyrate concentration.

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.

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.

Analysis of Pathotypes of Colletotrichum lindemuthianum Found in the Central Region of Mexico and Resistance in Elite Germ Plasm of Phaseolus vulgaris.

The pathotypes of 17 isolates of Colletotrichum lindemuthianum from the central region of Mexico were characterized to determine the genetic relationship among isolates from this region and other regions of Mexico, and to evaluate the resistance present in the elite germ plasm collection of Phaseolus vulgaris at INIFAP. Eight pathotypes were identified, including pathotype 292, which is reported for the first time in Mexico. The lack of isolates infecting cultivar TU carrying the Co-5 resistance gene suggests that this cultivar is a useful source of resistance. Six pathotypes produced susceptible reactions on only differential cultivars of Middle American origin, one pathotype on a single cultivar of Andean origin, and one pathotype on cultivars of both Middle American and Andean origin. Comparison of amplified fragment length polymorphism (AFLP) genotypes of 21 isolates confirmed suggestions that populations of C. lindemuthianum are comprised of asexually reproducing clonal lineages. Analysis of five different pathotypes of C. lindemuthianum on 21 elite genotypes of P. vulgaris identified four genotypes from different races of P. vulgaris resistant to all five pathotypes. This information will allow breeders and farmers to select the resistant genotypes most suited to their needs.