Karen A. Cichy

Saturation of an Intra-Gene Pool Linkage Map: Towards a Unified Consensus Linkage Map for Fine Mapping and Synteny Analysis in Common Bean

Map-based cloning and fine mapping to find genes of interest and marker assisted selection (MAS) requires good genetic maps with reproducible markers. In this study, we saturated the linkage map of the intra-gene pool population of common bean DOR364×BAT477 (DB) by evaluating 2,706 molecular markers including SSR, SNP, and gene-based markers. On average the polymorphism rate was 7.7% due to the narrow genetic base between the parents. The DB linkage map consisted of 291 markers with a total map length of 1,788 cM. A consensus map was built using the core mapping populations derived from inter-gene pool crosses: DOR364×G19833 (DG) and BAT93×JALO EEP558 (BJ). The consensus map consisted of a total of 1,010 markers mapped, with a total map length of 2,041 cM across 11 linkage groups. On average, each linkage group on the consensus map contained 91 markers of which 83% were single copy markers. Finally, a synteny analysis was carried out using our highly saturated consensus maps compared with the soybean pseudo-chromosome assembly. A total of 772 marker sequences were compared with the soybean genome. A total of 44 syntenic blocks were identified. The linkage group Pv6 presented the most diverse pattern of synteny with seven syntenic blocks, and Pv9 showed the most consistent relations with soybean with just two syntenic blocks. Additionally, a co-linear analysis using common bean transcript map information against soybean coding sequences (CDS) revealed the relationship with 787 soybean genes. The common bean consensus map has allowed us to map a larger number of markers, to obtain a more complete coverage of the common bean genome. Our results, combined with synteny relationships provide tools to increase marker density in selected genomic regions to identify closely linked polymorphic markers for indirect selection, fine mapping or for positional cloning.

Genome-wide association analysis of symbiotic nitrogen fixation in common bean

Key messageSignificant SNPs and candidate genes for symbiotic nitrogen fixation (SNF) and related traits were identified on Pv03, Pv07 and Pv09 chromosomes of common bean.AbstractA genome-wide association study (GWAS) was conducted to explore the genetic basis of variation for symbiotic nitrogen fixation (SNF) and related traits in the Andean Diversity Panel (ADP) comprising 259 common bean (Phaseolus vulgaris) genotypes. The ADP was evaluated for SNF and related traits in both greenhouse and field experiments. After accounting for population structure and cryptic relatedness, significant SNPs were identified on chromosomes Pv03, Pv07 and Pv09 for nitrogen derived from atmosphere (Ndfa) in the shoot at flowering, and for Ndfa in seed. The SNPs for Ndfa in shoot and Ndfa in seed co-localized on Pv03 and Pv09. Two genes Phvul.007G050500 and Phvul.009G136200 that code for leucine-rich repeat receptor-like protein kinases (LRR-RLK) were identified as candidate genes for Ndfa. LRR-RLK genes play a key role in signal transduction required for nodule formation. Significant SNPs identified in this study could potentially be used in marker-assisted breeding to accelerate genetic improvement of common bean for SNF.

Genome-Wide Association Study of Agronomic Traits in Common Bean

A genome‐wide association study (GWAS) using a global Andean diversity panel (ADP) of 237 genotypes of common bean (Phaseolus vulgaris L.) was conducted to gain insight into the genetic architecture of phenology, biomass, yield components, and seed yield traits. The panel was evaluated for 2 yr in field trials in Michigan and genotyped with 5398 single nucleotide polymorphism (SNP) markers. After correcting for population structure and cryptic relatedness, significant SNP markers associated with several agronomic traits were identified. Positional candidate genes, including Phvul.001G221100 on P. vulgaris (Pv) chromosome 01, associated with days to flowering and maturity were identified. Significant SNPs for seed yield were identified on Pv03 and Pv09 and colocalized with quantitative trait loci (QTL) for yield from previous studies conducted in several environments and contrasting genetic backgrounds. The majority of germplasm carrying the alleles with positive effects on seed yield was of African origin and largely underutilized in US breeding programs. The study provided insights into the genetic architecture of agronomic traits in Andean beans.

Effects of extrusion cooking on the chemical composition and functional properties of dry common bean powders

The impact of extrusion cooking on the chemical composition and functional properties of bean powders from four common bean varieties was investigated. The raw bean powders were extruded under eight different conditions, and the extrudates were then dried and ground (particle size⩽0.5mm). Compared with corresponding non-extruded (raw) bean powders (particle size⩽0.5mm), the extrusion treatments did not substantially change the protein and starch contents of the bean powders and showed inconsistent effects on the sucrose, raffinose and stachyose contents. The extrusion cooking did cause complete starch gelatinization and protein denaturation of the bean powders and thus changed their pasting properties and solvent-retention capacities. The starch digestibilities of the cooked non-extruded and cooked extruded bean powders were comparable. The extruded bean powders displayed functional properties similar to those of two commercial bean powders.

Induced mutants in common bean (Phaseolus vulgaris), and their potential use in nutrition quality breeding and gene discovery

Common bean (Phaseolus vulgaris) is the most widely grown grain legume for human consumption and a major protein and mineral source in East Africa and Latin America. It is also a simple diploid species with a small genome (650 Mb). Despite its nutritional and economic importance and tractable genome, P. vulgaris has a paucity of mutant resources compared to other crops, making it difficult to perform genetic screening in the species. In this review we discuss recent studies on mutagenesis that aim to produce large-scale, mutagenized populations for generalized trait screening, as well as previous EMS (ethyl methane sulfonate) and gamma radiation mutants that were developed for biological nitrogen fixation or plant morphology traits. Mutant stocks in this crop will allow researchers to conduct both forward (systematic phenotypic screening) and reverse genetics (such as TILLING, or Targeting Induced Local Lesions In Genomes) experiments aimed at understanding the genes involved in various traits, including abiotic and biotic stress tolerance, grain quality, and nutritional value, as well as genes involved in symbiosis with Rhizobia. Thus, mutant stocks will be important for gene discovery and creating novel variability. In this review, we highlight applications of mutation breeding for nutritional quality improvement of common bean, giving examples of seed protein, mineral content, and tannin accumulation traits.

The Phaseolus vulgaris ZIP gene family: identification, characterization, mapping, and gene expression

Zinc is an essential mineral for humans and plants and is involved in many physiological and biochemical processes. In humans, Zn deficiency has been associated with retarded growth and reduction of immune response. In plants, Zn is an essential component of more than 300 enzymes including RNA polymerase, alkaline phosphatase, alcohol dehydrogenase, Cu/Zn superoxidase dismutase, and carbonic anhydrase. The accumulation of Zn in plants involves many genes and characterization of the role of these genes will be useful in biofortification. Here we report the identification and phlyogenetic and sequence characterization of the 23 members of the ZIP (ZRT, IRT like protein) family of metal transporters and three transcription factors of the bZIP family in Phaseolus vulgaris L. Expression patterns of seven of these genes were characterized in two bean genotypes (G19833 and DOR364) under two Zn treatments. Tissue analyzed included roots and leaves at vegetative and flowering stages, and pods at 20 days after flowering. Four of the genes, PvZIP12, PvZIP13, PvZIP16, and Pv bZIP1, showed differential expression based on tissue, Zn treatment, and/or genotype. PvZIP12 and PvZIP13 were both more highly expressed in G19833 than DOR364. PvZIP12 was most highly expressed in vegetative leaves under the Zn (−) treatment. PvZIP16 was highly expressed in leaf tissue, especially leaf tissue at flowering stage grown in the Zn (−) treatment. Pv bZIP1 was most highly expressed in leaf and pod tissue. The 23 PvZIP genes and three bZIP genes were mapped on the DOR364 × G19833 linkage map. PvZIP12, PvZIP13, and PvZIP18, Pv bZIP2, and Pv bZIP3 were located near QTLs for Zn accumulation in the seed. Based on the expression and mapping results, PvZIP12 is a good candidate gene for increasing seed Zn concentration and increase understanding of the role of ZIP genes in metal uptake, distribution, and accumulation of zinc in P. vulgaris.

Transcriptome Characterization of Developing Bean (Phaseolus vulgaris L.) Pods from Two Genotypes with Contrasting Seed Zinc Concentrations

Dry bean (Phaseolus vulgaris L.) seeds are a rich source of dietary zinc, especially for people consuming plant-based diets. Within P. vulgaris there is at least two-fold variation in seed Zn concentration. Genetic studies have revealed seed Zn differences to be controlled by a single gene in two closely related navy bean genotypes, Albion and Voyager. In this study, these two genotypes were grown under controlled fertilization conditions and the Zn concentration of various plant parts was determined. The two genotypes had similar levels of Zn in their leaves and pods but Voyager had 52% more Zn in its seeds than Albion. RNA was sequenced from developing pods of both genotypes. Transcriptome analysis of these genotypes identified 27,198 genes in the developing bean pods, representing 86% of the genes in the P. vulgaris genome (v 1.0 DOE-JGI and USDA-NIFA). Expression was detected in 18,438 genes. A relatively small number of genes (381) were differentially expressed between Albion and Voyager. Differentially expressed genes included three genes potentially involved in Zn transport, including zinc-regulated transporter, iron regulated transporter like (ZIP), zinc-induced facilitator (ZIF) and heavy metal associated (HMA) family genes. In addition 12,118 SNPs were identified between the two genotypes. Of the gene families related to Zn and/or Fe transport, eleven genes were found to contain SNPs between Albion and Voyager.

Prediction of canned black bean texture (Phaseolus vulgaris L.) from intact dry seeds using visible/near-infrared spectroscopy and hyperspectral imaging data

BACKGROUND Texture is a major quality parameter for the acceptability of canned whole beans. Prior knowledge of this quality trait before processing would be useful to guide variety development by bean breeders and optimize handling protocols by processors. The objective of this study was to evaluate and compare the predictive power of visible and near infrared reflectance spectroscopy (visible/NIRS, 400-2498 nm) and hyperspectral imaging (HYPERS, 400-1000 nm) techniques for predicting texture of canned black beans from intact dry seeds. Black beans were grown in Michigan (USA) over three field seasons. The samples exhibited phenotypic variability for canned bean texture due to genetic variability and processing practice. Spectral preprocessing methods (i.e. smoothing, first and second derivatives, continuous wavelet transform, and two-band ratios), coupled with a feature selection method, were tested for optimizing the prediction accuracy in both techniques based on partial least squares regression (PLSR) models. RESULTS Visible/NIRS and HYPERS were effective in predicting texture of canned beans using intact dry seeds, as indicated by their correlation coefficients for prediction (Rpred ) and standard errors of prediction (SEP). Visible/NIRS was superior (Rpred = 0.546-0.923, SEP = 7.5-1.9 kg 100 g-1 ) to HYPERS (Rpred = 0.401-0.883, SEP = 7.6-2.4 kg 100 g-1 ), which is likely due to the wider wavelength range collected in visible/NIRS. However, a significant improvement was reached in both techniques when the two-band ratios preprocessing method was applied to the data, reducing SEP by at least 10.4% and 16.2% for visible/NIRS and HYPERS, respectively. Moreover, results from using the combination of the three-season data sets based on the two-band ratios showed that visible/NIRS (Rpred = 0.886, SEP = 4.0 kg 100 g-1 ) and HYPERS (Rpred = 0.844, SEP = 4.6 kg 100 g-1 ) models were consistently successful in predicting texture over a wide range of measurements. CONCLUSION Visible/NIRS and HYPERS have great potential for predicting the texture of canned beans; the robustness of the models is impacted by genotypic diversity, planting year and phenotypic variability for canned bean texture used for model building, and hence, robust models can be built based on data sets with high phenotypic diversity in textural properties, and periodically maintained and updated with new data.

Automated prediction of sensory scores for color and appearance in canned black beans (Phaseolus vulgaris L.) using machine vision

ABSTRACT Evaluation of canning quality of beans is commonly carried out by simple visual inspection that is time-consuming, resource intensive, and biased by the experience of the panelist. Moreover, there is not a standard scale to rate visual quality traits of canned beans. In this research, a machine vision system was implemented and tested for automatic inspection of color (COL) and appearance (APP) in canned black beans. Various color and textural image features (average, standard deviation, contrast, correlation, energy, and homogeneity from red, green, blue, lightness, red/green, yellow/blue, hue, saturation and value color scales) were extracted from beans and brine images, and evaluated to predict the quality rates for COL and APP of a group of bean panelists using multivariate statistics. Sixty-nine commercial canned black bean samples from different brands and markets were used for analysis. In spite of the “fair” agreement among the sensory panelists for COL and APP, as determined by multi-rater Kappa analysis, machine vision data based on partial least squares regression model showed high predictive performance for both COL and APP with correlation coefficients of 0.937 and 0.871, and standard errors of 0.26 and 0.38, respectively. When a classification was performed based on both COL and APP traits, a support vector machine model was able to sort the samples into two sensory quality categories of “acceptable” and “unacceptable” with an accuracy of 89.7%. Using simple color and texture image data, a machine vision system showed potential for the automatic evaluation of canned black beans by COL and/or appearance as a professional visual inspection.

Nutritional composition and cooking characteristics of tepary bean (Phaseolus acutifolius Gray) in comparison with common bean (Phaseolus vulgaris L.)

Tepary bean is a highly abiotic stress tolerant orphan crop for which there has been limited research on its nutritional value and cooking characteristics. These are key aspects when considering the potential for broader adoption of tepary bean. Therefore, the goal of this study was to evaluate a large set of seed composition and cooking traits related to human nutrition using both landraces and breeding lines of domesticated tepary bean from replicated field trials and to compare the traits in tepary with those in common bean. Tepary bean showed reduced fat and ash concentration and higher sucrose concentration as compared to common bean. Of the twelve amino acids evaluated, only proline in one of the two trials was statistically different between the two species. There were statistically significant differences between tepary and common bean for the concentration of some elements in this study; however, the elemental concentrations fell within the range of those found for common bean in previous studies. The majority of tepary bean lines showed consistently short cooking times and a high percentage of seeds showed measurable water uptake, while some showed a hardshell trait (low water uptake) and longer cooking times. Principal component analysis on a subset of traits showed a distinct group of common beans and two tepary bean groups that were divided on the basis of several agronomic, cooking, and elemental composition traits. Tepary bean, as with other pulses, is a highly nutritious crop with the range of composition and cooking characteristics similar to those of common bean. The variability for seed composition and cooking traits found within tepary bean can be exploited for its improvement.