Genetic dissection of yield-related traits in response to drought stress in common bean

Drought stress severely impairs common bean production.For facilitating drought-resistance breeding in common bean,molecular markers were identified in a genome-wide level marker–trait association study.A panel of 210 common bean accessions showed large variation in 11 agronomic traits at the adult stage(plant height,pod number per plant,seed number per pod,seed number per plant,seed yield per plant,pod length,harvest index,pod harvest index,days to maturity,hundred-seed weight,and seed yield)under two water conditions.The coefficient of variation ranged from 6.21% for pod harvest index to51.00% for seed number per plant under well-watered conditions,and from 4.05% for days to maturity to 40.72% for seed number per plant under drought stress.In a genome-wide association study,119quantitative-trait loci were associated with drought resistance,including 41 adjacent to known loci.Among these loci,12 were found to be associated with at least two traits.Three major loci were identified at Pv01 and Pv02.A set of candidate genes were found that encode MYBs,AREBs,WKRYs,and protein kinases.These results reveal promising alleles that control drought resistance,shedding light on the genetic basis of drought resistance and accelerating future efforts for drought resistance improvement in common bean.

Isolation and Authentication of Local Rhizobia Nodulating Common Bean (Phaseolus vulgaris L.) in Different Agro-Ecological Zones of Côte d’Ivoire

Legumes such as common bean (Phaseolus vulgaris L.) have been introduced into cropping systems for sustainable soil management. Consequently, the loss of fertility of the latter remains a major constraint to bean production because this legume is rarely fertilized, yet it is considered to be a poor nitrogen fixer in the absence of inoculation. To overcome this, this study was undertaken with the objective of seeking efficient local rhizobia in order to propose a bean inoculum formulation. To do this, soil samples taken from twelve localities in the Centre, North and West areas of C?te d’Ivoire were used to trap bean nodulating rhizobia. The ROBA1 bean accession used was sown in pots containing the sampled soils. Seedlings were uprooted at the start of flowering and nodulation was assessed. The isolates obtained were purified and then characterized phenotypically. The infectivity and symbiotic efficacy of these isolates were determined in vitro by the authentication test in which the purified isolates were reinoculated to their original host plant. A total of 24 rhizobium isolates were obtained from the soils of six localities. During morphological characterization, the isolates showed typical characteristics of Rhizobium. With the exception of RPC501, RPC505 and RPC522, all isolates were authenticated and able to nodulate the host plant in controlled culture. Isolates RPC502, RPC507, and RPC508 were effective and significantly increased (P < 0.05) nodule number and weight, height, and plant biomass. This study has, therefore, revealed the presence of effective local rhizobia in Ivorian soils and capable of nodulating common beans. A genetic characterization of efficient rhizobia identified after experimentation in different environmental conditions should be considered before being recommended as bean rhizobia inoculant.

Evaluation of common bean(Phaseolus vulgaris L.)genotypes for drought stress adaptation in Ethiopia

Drought stress linked with climate change is one of the major constraints faced by common bean farmers in Africa and elsewhere. Mitigating this constraint requires the selection of resilient varieties that withstand drought threats to common bean production.This study assessed the drought response of 64 small red-seeded genotypes of common bean grown in a lattice design replicated twice under contrasting moisture regimes,terminal drought stress and non-stress, in Ethiopia during the dry season from November2014 to March 2015. Multiple plant traits associated with drought were assessed for their contribution to drought adaptation of the genotypes. Drought stress determined by a drought intensity index was moderate(0.3). All the assessed traits showed significantly different genotypic responses under drought stress and non-stress conditions. Eleven genotypes significantly(P ≤ 0.05) outperformed the drought check cultivar under both drought stress and non-stress conditions in seed yielding potential. Seed yield showed positive and significant correlations with chlorophyll meter reading, vertical root pulling resistance force, number of pods per plant, and seeds per pod under both soil moisture regimes, indicating their potential use in selection of genotypes yielding well under drought stress and non-stress conditions. Clustering analysis using Mahalanobis distance grouped the genotypes into four groups showing high and significant inter-cluster distance, suggesting that hybridization between drought-adapted parents from the groups will provide the maximum genetic recombination for drought tolerance in subsequent generations.

Molecular cloning and characterization of a gene encoding the proline transporter protein in common bean(Phaseolus vulgaris L.)

As a typical compatible solute, proline is accumulated in plants under environmental stresses. Proline transporter(Pro T) plays an important role in proline distribution between plant organs. Using a candidate gene approach, we cloned a c DNA sequence for Pro T from common bean(Phaseolus vulgaris L.) and designated the gene Pv Pro T. The deduced amino acid sequence of Pv Pro T showed high similarity to Bet/Pro T proteins from other leguminous plants, and the highest similarity was observed with mothbean(Vigna aconitifolia L.) Vu Pro T.Relative quantification of the m RNA level of Pv Pro T using real-time PCR analysis showed that the Pv Pro T transcript level was higher in leaves than in stems and roots of common bean plants subjected to drought and salt stress. Under 20%(w/w) PEG-6000 treatment,drought-resistant plants expressed a higher level of Pv Pro T transcripts than droughtsensitive plants. Although heterologous expression of Pv Pro T in the Escherichia coli mutant mkh13 showed that Pv Pro T exhibited uptake activities for proline and betaine, no betaine content was detected in the common bean. These findings suggest that Pv Pro T plays an important role in the transportation of proline in common bean plants exposed to drought and salt stress.

QTL and candidate genes associated with common bacterial blight resistance in the common bean cultivar Longyundou 5 from China

Common bacterial blight(CBB), caused by Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans(Xff), is a worldwide disease of common bean(Phaseolus vulgaris L.).Longyundou 5, a Chinese cultivar in the Mesoamerican gene pool of common bean, displays resistance to the Xff strain XSC3-1. To identify the genetic mechanisms behind this resistance,we crossed Long 5 with a susceptible genotype to develop a mapping population of F2 plants.Plant resistance to CBB was identified at 14 and 21 days after inoculation with Xff strain XSC3-1.A major QTL at 14 and 21 days after inoculation was mapped on chromosome Pv10 with LOD scores of 6.41 and 5.35, respectively. This locus was associated with SAP6, a previouslyidentified and much-used dominant marker, but in a 4.2 cM interval between new codominant markers BMp10s174 and BMp10s244. Ten candidate genes were found between markers BMp10s174 and BMp10s244 on chromosome Pv10 and could encode defense response proteins responding to CBB pathogens. Four pairs each of epistatic QTL for CBB resistance were detected at 14 and 21 days after inoculation. Phenotypic variation explained by the epistatic QTL ranged from 7.19% to 12.15% and 7.72% to 8.80% at 14 and 21 days after inoculation, respectively. These results confirmed the importance of epistasis in CBB resistance in common bean. The adjacent markers found may be more efficient for marker assisted selection in common bean breeding for CBB resistance owing to their closer linkage to the target QTL.

Inheritance of marsh spot disease resistance in cranberry common bean(Phaseolus vulgaris L.)

Common bean(Phaseolus vulgaris) is an annual legume crop that is grown worldwide for its edible dry seeds and tender pods. Marsh spot(MS) of the seeds is a physio-genic stress disease affecting seed quality in beans. Studies have suggested that this disease involves a nutritional disorder caused by manganese deficiency, but the inheritance of resistance to this disease has not been reported. A biparental genetic population composed of 138 recombinant inbred lines(RILs) was developed from a cross between an MS resistant cultivar ‘Cran09’ and an MS susceptible cultivar ‘Messina’. The 138 RILs and their two parents were evaluated for MS resistance during five consecutive years from 2015 to 2019 in sandy and heavy clay soils in Morden, Manitoba, Canada. The MS incidence(MSI) and the MS resistance index(MSRI) representing disease severity were shown to be both highly correlated heritable traits that had high broad-sense heritability values(H;) of 86.5% and 83.2%, respectively. No significant differences for MSI and MSRI were observed between the two soil types in all five-(MSI) or four-year(MSRI) data collection, but significant correlations among years were observed despite MS resistance was moderately affected by year. The MSIs and MSRIs displayed a right-skewed distribution, indicating a mixed genetic model involving a few major genes and polygenes. Using the joint segregation analysis method, the same four major genes with additive-epistasis effects showed the best fit for both traits, explaining 84.4% and85.3% of the phenotypic variance for MSI and MSRI, respectively. For both traits, the M1, M2, M3 and m4 acted as the favorable(resistant) alleles for the four genes where M and m represent two alleles of each gene. However, due to epistatic effects, only the individuals of the M1 M2 M3 M4 haplotype appeared to be highly resistant, whereas those of the m1 m2 m3 M4 haplotype were the most susceptible. The m4 allele significantly suppressed the additive effects of M1 M2 M3 on resistance, but decreased susceptibility due to the additive effects of m1 m2 m3. Further quantitative trait locus(QTL) mapping is warranted to identify and validate individual genes and develop molecular markers for marker-assisted selection of resistant cultivars.

Molecular Marker-Assisted Backcrossing of Anthracnose Resistance Genes into Common Beans (<i>Phaseolus vulgaris</i>L.) Varieties

Anthracnose, caused by Colletotrichum lindemuthianum, is a major disease of common bean and results in high yield loss. Due to the high degree of pathogenic variability of the fungus and the continual emergence of new races, genetic resistance in the host is not durable. Gene pyramiding using Marker Assisted Selection (MAS) is proposed as a viable approach to improve the durability of major genes conditioning resistance to anthracnose. In this study a common bean line Urugezi x AND 1062 susceptible to anthracnose but already improved for Pythium root rot resistance was improved for anthracnose resistance through a backcross breeding program. Genotypic selection was done in Rubilizi laboratory in Kigali, Rwanada whereas phenotypic selection was conducted in an anthracnose hotspot at Rwerere, a research Centre of the Rwanda Agricultural and Animal Resources Development Board (RAB). Analysis of variance for effect of bean varieties and anthracnose isolates on disease expression showed significant differences (p < 0.001) among varieties and isolates and for the interaction between isolates and varieties. Developed BC2F1 plants were 41% of them resistant and 59% susceptible to anthracnose. However, the observed proportion of 26 resistants and 37 susceptible in BC2F1 plants didn’t fit the goodness of fit of the expected proportion of 75 resistants to 25 susceptible. Only 41% of BC2F1 plants inherited the resistance genes and were phenotypically resistant. Presence of SCAR-markers, SAB3 and SBB14, in the developed resistant lines h suggested successful resistance transfer of anthracnose resistance genes.

Exposure of Common Bean Seeds to Liquid Nitrogen Modifies Mineral Composition of Young Plantlet Leaves

Many publications describe cryopreservation techniques but only a few studies have focused on the biochemical and physiological changes occurring in plants regenerated from seeds exposed to liquid nitrogen. This paper aims at describing the effect of common bean seed cryostorage on mineral nutrition of young plantlets. The following elements were measured on leaves of 10-day-old plantlets from non-cryopreserved and cryopreserved seeds: Al, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, S, Se, Sr and Zn. At 10 days after sowing, both treatments (control and cryopreserved seeds) showed 100% seed germination without any visual phenotypic difference. However, contents of several elements in the leaves were different. Exposure of seeds to liquid nitrogen decreased Cu, Cd and Na uptake and increased absorption of B and Al. Further studies are required to understand the mechanisms underlying the relationship between seed exposure to liquid nitrogen and mineral nutrition during the early stages of plantlet growth.

Effect of Gamma Irradiation and Selection with Fungus Filtrate(Rhizoctonia solani Kuhn)on the in Vitro Culture of Common Bean(Phaseolus vulgaris)

The present investigation was undertaken to study the effect of gamma irradiation (dose from 10 to 100 Gy) and in vitro selection with fungus filtrate as selecting agent (concentration from 20% to 100%) on the susceptibility of the common bean to Rhizoctonia solani. The best results were found with a dose of 20 Gy or a concentration of 20% of fungus filtrate applied separately. These conditions were used to evaluate the combined effect of both approaches in a second experiment. The combined effect of irradiation and then selection adversely affected growth (height and roots) and survival of the in vitro plants. It may not be necessary to combine the variation generated by irradiation with the selection technique. For future assays we propose the application of: 1) gamma radiation, thereby inducing not only mutants with pathogen resistance, but also with other agronomic traits of interest. Later in the subculture MV4 potential fungus-resistant mutants will be evaluated in the field;or 2) selection pressure using fungus filtrate during three subcultures, which may be sufficient to induce the variation necessary to obtain in vitro plants resistant to fungus.

Solid State Bioconversion for Producing Common Bean(Phaseolus vulgaris L.)Functional Flour with High Antioxidant Activity and Antihypertensive Potential

The main objective of this investigation was to study the time effect during solid state bioconversion (SSB) on total phenolics content (TPC) and antioxidant activity (AoxA) of common beans to improve antihypertensive functionality. Cooked cotyledons of dehulled common beans were inoculated with a suspension of R. oligosporus NRRL 2710 (1 × 106 spores/mL), and incubated at 35℃ for times of 24, 36, 48, 60, 72, 84, 96 and 108 h (after 108 h the cotyledons showed off odor). Flours from bioprocessed dehulled common bean from each incubation time were blended with their corresponding milled seed coats. The best time for producing bioprocessed common bean (added with seed coats) functional flour with the highest AoxA (ORAC value = 17,468 μmol Trolox equivalents (TE)/100 gsample, dw;ABTS value = 13,505 μmol TE/100 gsample, dw) was 108 h. The SSB process substantially increased TPC and total hydrophilic AoxA and antihypertensive potential of common beans in 2.24, 2.45 - 2.73 and 6769 times, respectively. Proteins hydrolyzates from unprocessed whole and bioprocessed (108 h) common beans had IC50 [concentration needed to inhibit 50% the activity of angiotensin converting enzyme (ACE)] of 79.2 and 0.0117 μg/mL, respectively. The SSB is an efective strategy to improve the TPC of common beans for enhanced functionality with improved antioxidant activity and antihypertensive potential.

Comparisons of phaseolin type and α-amylase inhibitor in common bean(Phaseolus vulgaris L.)in China

The objective of this study was to characterize the phaseolin type and a-amylase(αAI) level in common bean(Phaseolus vidgaris L.) accessions deposited in the Chinese National Genebank.The 40 accessions sampled were common varieties originating in Asia,North America,South America,Europe,and Africa.No Inca(I-) phaseolin was observed in the accessions.Only four accessions contained Tendergreen(T-) phaseolin and the remaining36 contained Sanilac(S-) phaseolin.aAI proteins extracted from nine accessions showed higher a-amylase inhibitory activity than the control(Phase 2,IC_(50) = 0.65 μg).These common bean accessions have potential use as nutraceutical ingredients.

Phenolic Compound Profiles of Two Common Beans Consumed by Rwandans

Legumes are high-protein, medium-energy and micronutrient-rich food consumed in many parts of the world including Africa. This study evaluated the levels of specific phenolic compounds in three legumes. Two varieties of the common bean, (Phaseolus vulgaris L.) soybeans (Glycine max L.), and peas (Pisum sativum L.) from Rwanda were analyzed using high performance liquid chromatography with diode array detection. The phenolic compounds were identified by comparison to the chromatographic retention times and UV spectra of known reference compounds. This study results clearly shows the presence of 11 different phenolic compounds in common beans: gallic acid, (+)-catechin, (–)-epicatechin, caffeic acid, o-coumaric acid, chlorogenic acid, quercetin, 4-hydrobenzoic acid, syringic acid, ferulic acid and vanillic acid. The concentration ranged from 0.59 to 2.27 mg/kg for epicatechin. High levels of catechin (13.5 to 57.9 mg/kg) ferulic acid (26.1 to 47.6 mg/kg) were also observed. Therefore, the results of this study show that Rwandan common beans are a good source of phenolic acids in particular catechins and ferulic acid.

Common Bean Germplasm Diversity Study for Cold Tolerance in Ethiopia

Limited tolerance of common beans (Phaseolus vulgaris L.) to cold temperatures hinders an additional harvest during the small rainy season crop cycle (February to May) in the Ethiopian highlands that comprise two-thirds of the country. Therefore, identification of cold tolerant common bean genotypes is of paramount importance for the region. Field screening of 99 common bean genotypes originally procured from CIAT (International Center for Tropical Agriculture) was carried out for nine different plant traits associated with crop growth and yield at two locations differing in climates: Dire Dawa-higher daily maximum and minimum temperatures and absence of near zero chilling temperatures from February to May;Haramaya-lower daily maximum and minimum temperatures and occasionally near zero chilling temperatures during this period. The analysis of variance (ANOVA) showed the existence of significant variation among genotypes for the parameters measured. Principal component analysis (PCA) was carried out to assess the variation and correlation among genotypes for the traits and group them based on their performance at the two locations. The combination of first three principal components explained more than 50% of the genotypic variations. Principal component analysis was also able to discriminate the performance of genotypes between the two locations. It was grouped into at least 17 genotypes that were specific to Haramaya highland location. The results also revealed significant variation in performance among the 17 genotypes. These genotypes are specific to Ethiopian highlands and prominent resources for in-situ conservation of germplasms.

Hydrodynamic Modelling, Thermodynamic and Textural Variations during Common Beans Soaking

Hydrodynamic characteristics and its associated thermodynamic and textural variation of three common Malawian beans varieties (Boma, Sugar and Mandondo) during soaking were evaluated at four temperature regimes (25°C, 35°C, 45°C and 55°C). The equilibrium water uptake of 127% ± 5% was reached in 10, 6, and 4 hours respectively, for 25°C, 35°C and 45°C. Not much variation was observed between 45°C and 55°C except for sugar beans where equilibrium water uptake was reached within two hours of soaking at 55°C. Three models namely Peleg, two-parameter Mitscherlich model and viscoelastic model were used to evaluate the comparative predicting capabilities of the bean hydrodynamic characteristics. All models predicted the water absorption accurately (R2 > 0.903, RMSE activation kinetic parameters to be between 25 - 65 kJ/mol. Sugar beans were found to be the least hard. At room temperature, its hardness reduced by 58% within 2 hours of soaking. At higher temperature (55°C) hardness values were reduced to 12.5%, 11.1% and 15.0% within the first hour for Boma, Sugar and Mandondo beans, respectively.

Nitrate regulates rhizobial and mycorrhizal symbiosis in common bean(Phaseolus vulgaris L.)

Nitrogen-limited conditions are considered to be a prerequisite for legume-rhizobial symbiosis, but the effects of nitrate-rich conditions on symbiotic status remain poorly understood. We addressed this issue by examining rhizobial （Rhizobim tropici） and arbusclar mycorrhizal （Glomus intraradices） symbiosis in Phaseolus vulgaris L. cv. Negro Jamapa under nitrate pre-incubation and continuous nitrate conditions. Our results indicate that nitrate pre-incubation, independent of the concentration, did not affect nodule development. However, the continuous supply of nitrate at high concentrations impaired nodule maturation and nodule numbers. Low nitrate conditions, in addition to positively regulating nodule number, biomass, and nitrogenase activity, also extended the span of nitrogen-fixing activity. By contrast, for arbuscular mycorrhizae, continuous 1o and 50mmol/L nitrate increased the percent root length colonization, concomitantly reduced arbuscule size, and enhanced ammonia transport without affecting phosphate transport. Therefore, in this manuscript, we have proposed the importance of nitrate as a positive regulator in promoting both rhizobial and mycorrhizal symbiosis in the common bean.

The Diversity of Four Anti—nutritional Factors in Common Bean

Anti-nutritional factors such as lectins, saponin, trypsin inhibitor and phytic acid are endogenous substances in the common bean(Phaseolus vulgaris L.). In this study, the contents or activities of these anti-nutritional factors in fresh pods were detected in 56 selected cultivars. The results revealed significant difference within each factor in the tested cultivar population. The mean value of lectin content and the activity of trypsin inhibitor were 1.743 mg · g^(-1)and 1.680 mg · g^(-1)respectively. Their coefficients of variation(CV) were both more than 100% and each of the cultivar frequency distribution curve showed a main peak, but the discontinuous distributions in the extremely high and low areas indicate hierarchic cultivars. However, the mean contents of saponin and phytic acid were 3.730 mg · g^(-1)and 3.102 mg · g^(-1), respectively, with CV less than 41%.Each showed a main peak in its normal distribution curve and low frequency continuous distribution in dual tails. Meanwhile, statistic analysis demonstrated a positive correlation between the lectin content and trypsin inhibitor activity in fresh pods. Furthermore, all 56 tested cultivars were clustered into three groups based on their four anti-nutritional factor levels: 80% of them into medium level group, and 12% of them into low level group. The endogenous edible toxic compounds, such as lectin and trypsin inhibitor, are closely related to insect resistance in the field.This study suggests that it is possible to screen the cultivars containing less lectin and other factors but with reduced pest resistance in the field.

Efficacy of Metarhizium anisopliae in controlling the two-spotted spider mite Tetranychus urticae on common bean in screenhouse and field experiments

The efficacy of aqueous and emulsifiable formulations of the fungus Metarhizium anisopliae isolate ICIPE78 was evaluated on the population density of Tetranychus urticae infesting common bean plants under screenhouse and field conditions. Synthetic acaricide abamectin was included as a check. Bean plants were artificially infested with T. urticae and allowed to multiply. Three treatments were applied in the screenhouse and 1 treatment in field trials. Mite density was recorded 2 d before spraying and weekly postspraying. The number of pods per plant, number of seeds per pod, and the dry weight of seeds per plant were recorded only in the screenhouse trials. In both screenhouse and field trials, fungal formulations applied at the concentration of 108 conidia/mL and the aearicide reduced the population density of mites as compared to the controls. There were signif- icant differences in T. urticae population densities between the treatments at the various post-spraying sampling dates. In the screenhouse, the mite densities were near zero from 3-week postspraying in the treated leaves. At 4-week postspraying, there were no more leaves in the untreated control （T1） and in the control water ＋ Silwet-L77 （T2）. Fungal formulations were as effective as abamectin in reducing mite densities in both screenhouse and field experiments. There were significant differences in the production parameters during the 2 screenhouse trials, with fungal and abamectin treatments generally having the highest yield. Results of this study underline the potential of the M. anisopliae isolate ICIPE78 as an alternative to acaricides for T. urticae management.

Effects of low pH and aluminum stresses on common beans (Phaseolus vulgaris) differing in low-phosphorus and photoperiod responses

Using common beans differing greatly in the response to photoperiod and low-phosphorus(P)stress,we investigated their responses to acidity and aluminum(Al)toxicity and the relationship between Al tolerance and organic acid exudation under Al or low P stress.A genotype Ginshi was found to be sensitive to low pH treatment.When exposed to pH 4.5,serious curvature in the root tips of cv.Ginshi was observed;however,it was completely corrected by the application of 5 or 10μmol/L AlCl 3;increasing calcium(Ca)could ameliorate Al toxicity,but it could not correct root curvature at pH 4.5.Common beans showed significant differences in both root growth and Al tolerance,and the varieties from the Andes were more tolerant to Al toxicity than those from the Mesoamerican origin.In the presence of 50μmol/L AlCl 3,all the common bean genotypes exuded citrate,and a significant difference in the amounts of citrate was observed among genotypes.The genotypes originated in the Mesoamerica tended to release more citrate than other origins in the presence of Al.The P-inefficient genotype DOR364 exuded more citrate than the P-efficient genotype G19833 in the presence of 50μmol/L AlCl 3,whereas no organic acids were detected in root exudates under low-P stress.A reduction of citrate exudation in the DOR364,but a slight increase of citrate exudation in the G19833,was observed under Al stress after they were exposed to 6-d P starvation.These results suggest that different low-P or Al tolerance in common beans might not be associated with organic acid exudation.