Physiological Characteristics of Cultivated Tepary Bean (Phaseolus acutifolius A. Gray) and Its Wild Relatives Grown at High Temperature and Acid Soil Stress Conditions in the Amazon Region of Colombia

Common bean (Phaseolus vulgaris L.) is sensitive to different types of abiotic stresses (drought, high temperature, low soil fertility, and acid soil), and this may limit its adaptation and consequently to its yield under stress. Because of this, a sister species, tepary bean (Phaseolus acutifolius A. Gray), has recently gained attention in breeding for improved abiotic stress tolerance in common bean. In this study, we evaluated the adaptation of 302 accessions of tepary bean (Phaseolus acutifolius A. Gray) and its wild relatives (grouped in four types of tepary bean genetic resource: cultivated, acutifolius regressive, acutifolius wild, tenuifolius wild) when grown under high temperature and acid soil conditions with aluminum toxicity in the Amazon region of Colombia. Our objective was to determine differences among four types of tepary bean genetic resource in their morpho-phenological, agronomic, and physiological responses to combined high temperature and acid soil stress conditions. We found that cultivated P. acutifolius var acutifolius presented a greater number of pods per plant, as well as larger seeds and a greater number of seeds per pod. Some traits, such as root biomass, days to flowering and physiological maturity, specific leaf area, and stomatal density, showed significant differences between types of tepary bean genetic resource, probably contributing to difference in adaptation to combined stress conditions of high temperature and acid soil conditions. The photochemical quenching (qP) was higher in cultivated P. acutifolius var. acutifolius, while energy dissipation by non-photochemical quenching (NPQ) in the form of heat and the coefficient of non-photochemical dissipation (qN) were higher in acutifolius regressive and tenuifolius wild accessions. We have identified 6 accessions of cultivated and 19 accessions of tenuifolius wild that exhibited grain yields above 1800 kg ha−1. These accessions could be suitable to use as parents to improve dry seed production of tepary bean under combined stress conditions of high temperature and acid soil.

Heat stress-induced changes in shoot and root characteristics of genotypes of tepary bean (Phaseolus acutifolius A. Gray), common bean (Phaseolus vulgaris L.) and their interspecific lines

Heat stress is a major limitation to grain yield in common bean (Phaseolus vulgaris L.). Tepary bean (Phaseolus acutifolius A. Gray) is better adapted to heat stress than common bean. Ten tepary bean accessions, four common bean genotypes and four interspecific lines involving P. vulgaris and P. acutufolius, P. coccineus and P. dumosus were evaluated for tolerance to heat stress conditions induced under greenhouse conditions and these were compared to plants grown under ambient temperatures. The high temperature treatment was 29 ±5 °C during the day and was >24 °C (up to 27 °C) during the night, while the ambient temperature (AT) treatment was 25 ±5 °C during the day and 19± 2 °C at night. The genotypic differences were evaluated for morpho-physiological characteristics of shoot and root and also yield components. The Genotype and Genotype × Temperature interactions were significant for all shoot and root morpho-physiological characteristics evaluated. Higher temperature (HT) significantly affected leaf photosynthetic efficiency, total chlorophyll content, and stomatal conductance. The effect was positive or negative, depending on the genotypes. Tepary accessions showed reduced total chlorophyll content, while common bean genotypes and the interspecific lines were less affected. Tepary accessions also showed reduced stomatal conductance, but increased leaf photosynthetic efficiency under HT. Common bean genotypes increased stomatal conductance and decreased leaf photosynthetic efficiency. High temperature decreased total root length, specific root length and pod biomass compared to ambient conditions, but there was no marked effect on pollen viability of the tested genotypes. The superior adaptation of tepary germplasm accessions to high temperature is attributed to their ability to regulate stomatal opening and photosynthetic efficiency, together with a superior ability to remobilize photosynthates from older leaves to pods during physiological maturity

Mutation breeding for heat and drought tolerance in tepary bean (Phaseolus acutifolius A. Gray)

Tepary bean (Phaseolus acutifolius A. Gray) is more heat and drought tolerant than common bean (P. vulgaris L.). Four hundred mutant lines of two tepary accessions (G40068 and G40159) were generated by ethyl methane sulfonate (EMS) treatment. In preliminary studies of the M5 mutant lines under abiotic stress, three mutant lines (CMT 38, CMT 109, CMT 187) were selected from six mutated lines based on morpho-physiological traits and superior yield and advanced to the M6 generation. The M6 mutant lines were uniform and genetically stable. These mutant lines and their original (M0) parents were evaluated for heat and drought tolerance under greenhouse conditions. Their performance was evaluated for morpho-physiological attributes, seed yield and yield components. Under high temperature and drought conditions, the CMT 38 mutant (M6 line) and its original tepary (M0) accession (G40068) showed greater values of pod biomass, pod number and 100-seed biomass than the other lines tested. The CMT 109 and CMT 187 mutant lines and their G40159 original accession (M0) also showed the highest value of seed number under high temperature and drought conditions. This suggests that the previous screening performed during the population advancement of these mutant lines, based on morphological traits like growth habit, was not detrimental to the yield variables evaluated here. Under combined heat and drought conditions, different parameters could be incorporated into tepary breeding programmes, as selection criteria to screen genotypes for tolerance to heat and drought stress. These parameters included: chlorophyll (SPAD) readings, seed biomass, 100-seed biomass and seed number because they explain the observed variance in the principal component analysis. Two additional traits (root biomass and stem diameter) were also identified as useful attributes, based on univariate analysis. The mutant lines evaluated here offer potential for further improvement of tepary bean to high temperature and drought

Protein Concentrates on Tepary Bean (Phaseolus acutifolius Gray) as a Functional Ingredient: In silico Docking of Tepary Bean Lectin to Peroxisome Proliferator-Activated Receptor Gamma

The tepary bean (Phaseolus acutifolius Gray) is a US–Mexico frontier native crop, produces high yields in agriculture, and needs to be reconsidered because of its nutritional and functional properties. This study aimed to determine the technological and nutritional properties of flours and protein concentrates of tepary bean, besides determining an in silico agonist effect of tepary bean lectin to peroxisome proliferator-activated receptor gamma (PPAR-γ). We evaluated the technological properties of raw samples (tepary flour and tepary protein concentrate) and cooked samples (tepary flour and tepary protein concentrate). The flours present a significant difference (p < 0.05) concerning protein concentrates in water absorption and oil absorption capacity. The raw samples' emulsifying capacity was higher than that reported in the literature for other legumes, but not the cooked samples. The samples' foaming capacity had no significant difference in treatments (p > 0.05), and cooked tepary bean protein concentrate presented complete gelation at a lower concentration (2%). Nutritionally, raw samples present a protein percentage of 23.46 ± 0.06 and 71.38 ± 0.44 and cooked samples present a protein percentage of 25.27 ± 0.04 and 62.69 ± 0.14; a chemical score of 72, 86, 82, and 72; in vitro protein digestibility (%) = 48.20 ± 0.31, 49.80 ± 0.80, 61.77 ± 1.70, and 63.61 ± 4.19; and C-PER = 0.86, 1.34, 1.93, and 1.81, respectively. All the samples showed methionine + cysteine as the limiting amino acid. All these nutritional data are very similar to the common bean (Phaseolus vulgaris). SDS-PAGE preserves the lectin fraction in both protein concentrates. The in silico study of tepary lectin (PDB: 6tt9) shows that there were seven peptides that presented values below −120 kcal/mol: PEW, VSVGF, PSQK, TTPW, ATSF, ITY, and TSF, with VSVGF, PSQK, and PEW having the highest affinity for active sites of the PAPRγ receptor (binding energies from −5.32 to −7.04 kcal/mol). These peptides could show antiadipogenic or antidiabetic activity based on the intermolecular bond energies and open an interesting research item.

Allelic Diversity at Abiotic Stress Responsive Genes in Relationship to Ecological Drought Indices for Cultivated Tepary Bean, Phaseolus acutifolius A. Gray, and Its Wild Relatives

Some of the major impacts of climate change are expected in regions where drought stress is already an issue. Grain legumes are generally drought susceptible. However, tepary bean and its wild relatives within Phaseolus acutifolius or P. parvifolius are from arid areas between Mexico and the United States. Therefore, we hypothesize that these bean accessions have diversity signals indicative of adaptation to drought at key candidate genes such as: Asr2, Dreb2B, and ERECTA. By sequencing alleles of these genes and comparing to estimates of drought tolerance indices from climate data for the collection site of geo-referenced, tepary bean accessions, we determined the genotype x environmental association (GEA) of each gene. Diversity analysis found that cultivated and wild P. acutifolius were intermingled with var. tenuifolius and P. parvifolius, signifying that allele diversity was ample in the wild and cultivated clade over a broad sense (sensu lato) evaluation. Genes Dreb2B and ERECTA harbored signatures of directional selection, represented by six SNPs correlated with the environmental drought indices. This suggests that wild tepary bean is a reservoir of novel alleles at genes for drought tolerance, as expected for a species that originated in arid environments. Our study corroborated that candidate gene approach was effective for marker validation across a broad genetic base of wild tepary accessions.

Bioaccessibility and In Vitro Intestinal Permeability of a Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) Using the Everted Intestine Assay

Tepary bean (Phaseolus acutifolius) lectins exhibit differential in vitro and in vivo biological effects, but their gastrointestinal interactions and digestion have not yet been assessed. This work aimed to evaluate the changes of a recombinant Tepary bean lectin (rTBL-1) through an in vitro and ex vivo gastrointestinal process. A polyclonal antibody was developed to selectively detect rTBL-1 by Western blot (WB) and immunohistochemical analysis. Everted gut sac viability was confirmed until 60 min, where protein bioaccessibility, apparent permeability coefficient, and efflux ratio showed rTBL-1 partial digestion and absorption. Immunoblot assays suggested rTBL-1 internalization, since the lectin was detected in the digestible fraction. The immunohistochemical assay detected rTBL-1 presence at the apical side of the small intestine, potentially due to the interaction with the intestinal cell membrane. The in silico interactions between rTBL-1 and some saccharides or derivatives showed high binding affinity to sialic acid (−6.70 kcal/mol) and N-acetylglucosamine (−6.10 kcal/mol). The ultra-high-performance liquid chromatography–electron spray ionization–quantitative time-of-flight coupled to mass spectrometry (UHPLC–ESI–QTOF/MS) analysis showed rTBL-1 presence in the gastric content and the non-digestible fraction after intestinal simulation conditions. The results indicated that rTBL-1 partially resisted the digestive conditions and interacted with the intestinal membrane, whereas its digestion allowed the absorption or internalization of the protein or the derivative peptides. Further purification of digestion samples should be conducted to identify intact rTBL-1 protein and digested peptides to assess their physiological effects.

Effect of Plant Pattern and Density on Yield and Amino Acid Profile of Tepary Bean (Phaseolus acutifolius) in a Semiarid Area

Background: Tepary bean is recognized as a deal crop for cultivation in arid and semi-arid regions. The spatial distribution of plants is an important farming practice. A study was conducted in tepary bean (Phaseolus acutifolius A. Gray) to investigate the effect of planting pattern and plant density on plant yield and amino acid profile.Methods: The experiment was carried out in a split-split plot design with three replications during 2017 and 2018. The main plot comprised planting pattern at two levels, including one-row plantation (PP1) and two-row plantation (PP2) and sub plot comprised plant density at three levels, including high plant density (HPD:50 plant/m2), moderate plant density (MPD:40 plant/m2) and low plant density (LPD: 30 plant/m2). In both years, plants were harvested at the end of pod filling stage at October. Amino acid profiles were determined using a HPLC system. Result: The results showed that in the first year and under PP2, MPD increased the biological yield by 19% and 16% compared to HPD and LPD, respectively. Increased pod yield was observed at MPD compared to HPD and LPD. In the second year under PP1, MPD improved grain yield by 75% and 65% in comparison to HPD and LPD, respectively. Increased protein and amino acid content were observed at MPD and PP2. The greatest amino acid content was Leu followed by Lys, Arg, Phe, Val, Ile, Thr, Tyr, His, Met, Trp and Cys.

Tepary bean (Phaseolus acutifolius) lectin fraction provokes reversible adverse effects on rats’ digestive tract

The Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) exhibits differential cytotoxicity on colon cancer cells and inhibition of early tumorigenesis in the colon (50 mg/kg, three times per week, for 6 weeks). TBLF showed low toxicity with the ability to activate the immune system; however, some adverse effects are the loss in body weight gain, intestinal atrophy, and pancreatic hyperplasia. After a recovery period of 2 weeks after treatment, reversion of pancreatic hyperplasia but no recovery of intestinal atrophy was observed. As TBLF has shown anticancer effects on the colon, it is important to characterize the adverse effects and how they can be reversed. Sprague Dawley rats were administered with TBLF (50 mg/kg) for 6 weeks, three times per week, and then allowed to recover for 6 weeks post-treatment. After TBLF administration, small intestine atrophy, villus atrophy, and cryptic hyperplasia were confirmed, as well as increased intestinal mucus production, increased permeability and a decrease in the apparent ileal digestibility of crude proteins. The colon showed damage in the simple prismatic tissue and decreased crypt depth, and changes in microbiota and a decrease in the apparent fecal digestibility of crude protein were determined. Our results show that the adverse effects provoked by TBLF were partially reversed after 6 weeks of recovery post-treatment, suggesting that increasing the recovery period it could be possible to reverse all adverse effects observed.