Polyphenolic compounds appear to limit the nutritional benefit of biofortified higher iron black bean (Phaseolus vulgaris L.).

BACKGROUND: Our objective was to determine if a biofortified variety of black bean can provide more bioavailable-iron (Fe) than a standard variety. Two lines of black beans (Phaseolus-vulgaris L.), a standard (DOR500; 59µg Fe/g) and biofortified (MIB465; 88µg Fe/g) were used. The DOR500 is a common commercial variety, and the MIB465 is a line developed for higher-Fe content. Given the high prevalence of Fe-deficiency anemia worldwide, it is important to determine if Fe-biofortified black beans can provide more absorbable-Fe. METHODS: Black bean based diets were formulated to meet the nutrient requirements for the broiler (Gallus-gallus) except for Fe (dietary Fe-concentrations were 39.4±0.2 and 52.9±0.9 mg/kg diet, standard vs. biofortified, respectively). Birds (n=14) were fed the diets for 6-weeks. Hemoglobin-(Hb), liver-ferritin and Fe-related transporter/enzyme gene-expression were measured. Hemoglobin-maintenance-efficiency and total-body-Hb-Fe values were used to estimate Fe-bioavailability. RESULTS: Hemoglobin-maintenance-efficiency values were higher (P<0.05) in the group consuming the standard-Fe beans on days 14, 21 and 28; indicating a compensatory response to lower dietary-Fe. Final total-Hb-Fe body content was higher in the biofortified vs. the standard group (26.6±0.9 and 24.4±0.8 mg, respectively; P<0.05). There were no differences in liver-ferritin or in expression of DMT-1, Dcyt-B, and ferroportin. In-vitro Fe-bioavailability assessment indicated very low Fe-bioavailability from both diets and between the two bean varieties (P>0.05). Such extremely-low in-vitro Fe-bioavailability measurement is indicative of the presence of high levels of polyphenolic-compounds that may inhibit Fe-absorption. High levels of these compounds would be expected in the black bean seed-coats. CONCLUSIONS: The parameters of Fe-status measured in this study indicate that only a minor increase in absorbable-Fe was achieved with the higher-Fe beans. The results also raise the possibility that breeding for increased Fe-concentration elevated the levels of polyphenolic-compounds that can reduce bean Fe-bioavailability, although the higher levels of polyphenolics in the higher-Fe beans may simply be coincidental or an environmental effect. Regardless, Fe-biofortified beans remain a promising vehicle for increasing intakes of bioavailable-Fe in human populations that consume high levels of these beans as a dietary staple, and the bean polyphenol profile must be further evaluated and modified if possible in order to improve the nutritional quality of higher-Fe beans.

QTL analyses for seed iron and zinc concentrations in an intra-genepool population of Andean common beans (Phaseolus vulgaris L.).

Legumes provide essential micronutrients that are found only in low amounts in the cereals or root crops. An ongoing project at CIAT has shown that the legume common bean is variable in the amount of seed minerals (iron, zinc, and other elements), vitamins, and sulfur amino acids that they contain and that these traits are likely to be inherited quantitatively. In this study we analyzed iron and zinc concentrations in an Andean recombinant inbred line (RIL) population of 100 lines derived from a cross between G21242, a Colombian cream-mottled climbing bean with high seed iron/zinc and G21078, an Argentinean cream seeded climbing bean with low seed iron/zinc. The population was planted across three environments; seed from each genotype was analyzed with two analytical methods, and quantitative trait loci (QTL) were detected using composite interval mapping and single-point analyses. A complete genetic map was created for the cross using a total of 74 microsatellite markers to anchor the map to previously published reference maps and 42 RAPD markers. In total, nine seed mineral QTL were identified on five linkage groups (LGs) with the most important being new loci on b02 and other QTL on b06, b08, and b07 near phaseolin. Seed weight QTL were associated with these on b02 and b08. These Andean-derived QTL are candidates for marker-assisted selection either in combination with QTL from the Mesoamerican genepool or with other QTL found in inter and intra-genepool crosses, and the genetic map can be used to anchor other intra-genepool studies.

QTL for seed iron and zinc concentration and content in a Mesoamerican common bean (Phaseolus vulgaris L.) population.

Iron and zinc deficiencies are human health problems found throughout the world and biofortification is a plant breeding-based strategy to improve the staple crops that could address these dietary constraints. Common bean is an important legume crop with two major genepools that has been the focus of genetic improvement for seed micronutrient levels. The objective of this study was to evaluate the inheritance of seed iron and zinc concentrations and contents in an intra-genepool Mesoamerican × Mesoamerican recombinant inbred line population grown over three sites in Colombia and to identify quantitative trait loci (QTL) for each mineral. The population had 110 lines and was derived from a high-seed iron and zinc climbing bean genotype (G14519) crossed with a low-mineral Carioca-type, prostrate bush bean genotype (G4825). The genetic map for QTL analysis was created from SSR and RAPD markers covering all 11 chromosomes of the common bean genome. A set of across-site, overlapping iron and zinc QTL was discovered on linkage group b06 suggesting a possibly pleiotropic locus and common physiology for mineral uptake or loading. Other QTL for mineral concentration or content were found on linkage groups b02, b03, b04, b07, b08 and b11 and together with the b06 cluster were mostly novel compared to loci found in previous studies of the Andean genepool or inter-genepool crosses. The discovery of an important new locus for seed iron and zinc concentrations may facilitate crop improvement and biofortification using the high-mineral genotype especially within the Mesoamerican genepool.

Single and Multi-trait GWAS Identify Genetic Factors Associated with Production Traits in Common Bean Under Abiotic Stress Environments.

The genetic improvement of economically important production traits of dry bean (Phaseolus vulgaris L.), for geographic regions where production is threatened by drought and high temperature stress, is challenging because of the complex genetic nature of these traits. Large scale SNP data sets for the two major gene pools of bean, Andean and Middle American, were developed by mapping multiple pools of genotype-by-sequencing reads and identifying over 200k SNPs for each gene pool against the most recent assembly of the P. vulgaris genome sequence. Moderately sized B ean A biotic S tress E valuation (BASE) panels, consisting of genotypes appropriate for production in Central America and Africa, were assembled. Phylogenetic analyses demonstrated the BASE populations represented broad genetic diversity for the appropriate races within the two gene pools. Joint mixed linear model genome-wide association studies with data from multiple locations discovered genetic factors associated with four production traits in both heat and drought stress environments using the BASE panels. Pleiotropic genetic factors were discovered using a multi-trait mixed model analysis. SNPs within or near candidate genes associated with hormone signaling, epigenetic regulation, and ROS detoxification under stress conditions were identified and can be used as genetic markers in dry bean breeding programs.

Studies of Cream Seeded Carioca Beans (Phaseolus vulgaris L.) from a Rwandan Efficacy Trial: In Vitro and In Vivo Screening Tools Reflect Human Studies and Predict Beneficial Results from Iron Biofortified Beans.

Iron (Fe) deficiency is a highly prevalent micronutrient insufficiency predominantly caused by a lack of bioavailable Fe from the diet. The consumption of beans as a major food crop in some populations suffering from Fe deficiency is relatively high. Therefore, our objective was to determine whether a biofortified variety of cream seeded carioca bean (Phaseolus vulgaris L.) could provide more bioavailable-Fe than a standard variety using in-vivo (broiler chicken, Gallus gallus) and in-vitro (Caco-2 cell) models. Studies were conducted under conditions designed to mimic the actual human feeding protocol. Two carioca-beans, a standard (G4825; 58 µg Fe/g) and a biofortified (SMC; 106 µg Fe/g), were utilized. Diets were formulated to meet the nutrient requirements of Gallus gallus except for Fe (33.7 and 48.7 µg Fe/g, standard and biofortified diets, respectively). In-vitro observations indicated that more bioavailable-Fe was present in the biofortified beans and diet (P<0.05). In-vivo, improvements in Fe-status were observed in the biofortified bean treatment, as indicated by the increased total-body-Hemoglobin-Fe, and hepatic Fe-concentration (P<0.05). Also, DMT-1 mRNA-expression was increased in the standard bean treatment (P<0.05), indicating an upregulation of absorption to compensate for less bioavailable-Fe. These results demonstrate that the biofortified beans provided more bioavailable Fe; however, the in vitro results revealed that ferritin formation values were relatively low. Such observations are indicative of the presence of high levels of polyphenols and phytate that inhibit Fe absorption. Indeed, we identified higher levels of phytate and quercetin 3-glucoside in the Fe biofortified bean variety. Our results indicate that the biofortified bean line was able to moderately improve Fe-status, and that concurrent increase in the concentration of phytate and polyphenols in beans may limit the benefit of increased Fe-concentration. Therefore, specific targeting of such compounds during the breeding process may yield improved dietary Fe-bioavailability. Our findings are in agreement with the human efficacy trial that demonstrated that the biofortified carioca beans improved the Fe-status of Rwandan women. We suggest the utilization of these in vitro and in vivo screening tools to guide studies aimed to develop and evaluate biofortified staple food crops. This approach has the potential to more effectively utilize research funds and provides a means to monitor the nutritional quality of the Fe-biofortified crops once released to farmers.

Development of a Mesoamerican intra-genepool genetic map for quantitative trait loci detection in a drought tolerant × susceptible common bean (Phaseolus vulgaris L.) cross.

Drought is a major constraint to common bean (Phaseolus vulgaris L.) production, especially in developing countries where irrigation for the crop is infrequent. The Mesoamerican genepool is the most widely grown subdivision of common beans that include small red, small cream and black seeded varieties. The objective of this study was to develop a reliable genetic map for a Mesoamerican × Mesoamerican drought tolerant × susceptible cross and to use this map to analyze the inheritance of yield traits under drought and fully irrigated conditions over 3 years of experiments. The source of drought tolerance used in the cross was the cream-seeded advanced line BAT477 crossed with the small red variety DOR364 and the population was made up of recombinant inbred lines in the F5 generation. Quantitative trait loci were detected by composite interval mapping for the traits of overall seed yield, yield per day, 100 seed weight, days to flowering and days to maturity for each field environment consisting of two treatments (irrigated and rainfed) and lattice design experiments with three repetitions for a total of six environments. The genetic map based on amplified fragment length polymorphism and random amplified polymorphic DNA markers was anchored with 60 simple sequence repeat (SSR) markers and had a total map length of 1,087.5 cM across 11 linkage groups covering the whole common bean genome with saturation of one marker every 5.9 cM. Gaps for the genetic map existed on linkage groups b03, b09 and b11 but overall there were only nine gaps larger than 15 cM. All traits were inherited quantitatively, with the greatest number for seed weight followed by yield per day, yield per se, days to flowering and days to maturity. The relevance of these results for breeding common beans is discussed in particular in the light of crop improvement for drought tolerance in the Mesoamerican genepool.

Corrigendum to: Genetic mapping of basal root gravitropism and phosphorus acquisition efficiency in common bean.

Root gravitropism determines the relative distribution of plant roots in different soil layers, and therefore, may influence the acquisition of shallow soil resources such as phosphorus (P). Growth pouch and field studies were conducted to evaluate root gravitropism of common bean (Phaseolus vulgaris L.) in response to P deficiency and to detect quantitative trait loci (QTL) associated with this trait. A deep-rooted genotype, DOR364, was crossed with a shallow-rooted genotype, G19833, to obtain 86 F5.7 recombinant inbred lines (RILs). Root gravitropic traits were measured as basal root growth angle (BRGA), shallow basal root length (SBRL, basal root length in the top 0-3 cm of soil) and relative shallow basal root length (RSBRL, percentage of basal root length in the top 0-3 cm of soil relative to total basal root length). Large genetic variability for these traits was found in the parents and RILs, with BRGA ranging from -18.73 to 56.69º and SBRL ranging from 0.42 to 2.63 m per plant. The parents and six RILs with contrasting root gravitropism were further evaluated in the field, where root shallowness was significantly correlated with plant growth and P uptake. QTL were detected by single point analysis (SPA), interval mapping (IM) and composite interval mapping (CIM) techniques with a genetic map for the DOR364 × G19833 population consisting of 236 molecular markers. The IM/CIM QTL were detected among the 11 linkage groups of common bean, with 16 QTL controlling the above root traits and six QTL controlling P acquisition efficiency (PAE) in the field study. At least three of the root trait QTL were associated with QTL for PAE, suggesting that root gravitropic traits are associated with PAE and that QTL for these traits can be used to facilitate selection and breeding for higher P efficiency in common bean and other crops.

Genetic mapping of basal root gravitropism and phosphorus acquisition efficiency in common bean.

Root gravitropism determines the relative distribution of plant roots in different soil layers, and therefore, may influence the acquisition of shallow soil resources such as phosphorus (P). Growth pouch and field studies were conducted to evaluate root gravitropism of common bean (Phaseolus vulgaris L.) in response to P deficiency and to detect quantitative trait loci (QTL) associated with this trait. A deep-rooted genotype, DOR364, was crossed with a shallow-rooted genotype, G19833, to obtain 86 F5.7 recombinant inbred lines (RILs). Root gravitropic traits were measured as basal root growth angle (BRGA), shallow basal root length (SBRL, basal root length in the top 0-3 cm of soil) and relative shallow basal root length (RSBRL, percentage of basal root length in the top 0-3 cm of soil relative to total basal root length). Large genetic variability for these traits was found in the parents and RILs, with BRGA ranging from -18.73 to 56.69º and SBRL ranging from 0.42 to 2.63 m per plant. The parents and six RILs with contrasting root gravitropism were further evaluated in the field, where root shallowness was significantly correlated with plant growth and P uptake. QTL were detected by single point analysis (SPA), interval mapping (IM) and composite interval mapping (CIM) techniques with a genetic map for the DOR364 × G19833 population consisting of 236 molecular markers. The IM / CIM QTL were detected among the 11 linkage groups of common bean, with 16 QTL controlling the above root traits and six QTL controlling P acquisition efficiency (PAE) in the field study. At least three of the root trait QTL were associated with QTL for PAE, suggesting that root gravitropic traits are associated with PAE and that QTL for these traits can be used to facilitate selection and breeding for higher P efficiency in common bean and other crops.