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George L. Hosfield and Clifford W. Beninger

Seed coat color in dry bean (Phaseolus vulgaris L.) is determined by the presence or absence of tannins, flavonoids, and anthocyanins. Black beans contain three main anthocyanins that are responsible for their black seed coat color: delphinidin 3-O-glucoside, petunidin 3-O-glucoside, and malvidin 3-O-glucoside. Leaching of anthocyanins occurs in many black bean genotypes during thermal processing (i.e., blanching and cooking). Black beans that lose their dark color after processing are unacceptable to the industry. Since the marketability of black beans can be adversely affected by thermal processing, an experiment was conducted to ascertain whether pigment leaching was due to qualitative or quantitative changes in anthocyanins during processing. Four black bean genotypes that showed differential leaching of color were investigated. `Harblack' retains most of its black color after processing while `Raven' loses most of its color. `Black Magic' and `Black Jack' are intermediate between `Harblack' and `Raven' in processed color. Bean samples (119 ± 1.5 g) of the four genotypes were thermally processed in 100 x 75-mm tin cans in a pilot laboratory. Seed coats were removed from the cooked beans, freeze-dried, and placed in solutions of formic 10 acid: 65 water: 25 methanol to extract anthocyanins. The extracts were analyzed by HPLC. Although all genotypes retained some color, there were no detectable anthocyanins in seed coats of the cooked beans. In a second experiment, raw beans of each genotype were boiled in distilled water for 15 minutes. All four genotypes lost color during boiling, but `Harblack' retained most of its color and had a five-fold higher concentration of the three anthocyanins than did the other genotypes. `Harblack' may retain color better than other black beans because of physical characteristics of the seed coat.

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Krista C. Shellie and George L. Hosfield

Genetic and environmental interactions for bean cooking time, water absorption, and protein content were estimated with 10 dry bean (Phaseolus vulgaris L.) cultivars grown at three locations in Rwanda, Africa, during five consecutive harvests. The genotypic variance component was larger than genotype × environment variance components for the cooking time index and percent water absorption. No significant genotypic effect was observed for seed protein content. The phenotypic correlation (-0.37) between the cooking time index and percent water absorption was not strong enough to justify the use of water absorption as an indirect selection method for cooking time. The most efficient allocation of resources to evaluate the cooking time of common bean cultivars with a 25-pin bar-drop cooker was four field replications over two harvests at two locations. Water absorption was evaluated most efficiently with four field replications over two harvests at a single location.

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Kimberly J Walters, George L. Hosfield and James D. Kelly

Ninety-eight percent of the navy beans (Phaseolus vulgaris) grown in the US are processed. Thus, new cultivars considered for release must meet industry standards. Canning quality behaves as a classical QTL which precludes its selection and evaluation in early generations. Such delays add a measure of inefficiency to a breeding program. Indirect selection for canning quality using molecular markers could increase efficiency. RAPD markers are more useful than RFLP's, in Phaseolus, due to a simpler protocol and a higher level of polymorphism within genetically related cultivars. Three populations of RIL's, derived from crosses between cultivars with standard and sub-standard canning quality, were screened to identify markers associated with canning quality. Material for evaluation was grown at two locations, in three replications and processed, in the Food Science Processing Lab, following industry standards. Quality traits measured were: processed texture, color and appearance. Associations of putative markers with canning quality were identified using ANOVA and Mapmaker programs

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Clifford W. Beninger, George L. Hosfield and Muraleedharan Nair

Dry bean (Phaseolus vulgaris L.) seed coat color is determined by the presence and relative amounts of phenolics, flavonoids, and anthocyanins present in the lumen of epidermal cells. Some of these chemicals may interact with proteins of the cotyledon to form complexes that render beans hard to cook and digest. Eight genetic loci control seed pigment chemistry. When all eight loci are dominant, a shiny black seed coat results, but recessive substitutions at one or more loci yield colors ranging from white, yellow, and brown to dark violet. In order to relate Mendelian genes for seed coat color to the pigments formed, we studied eight genetic stocks that had recessive substitutions at one or more color-determining loci in an otherwise all-dominant genetic background. Seed coat from each genotype was extracted exhaustively with hexane, EtOAc, MeOH, MeOH:H2O 1:1, and H2O 100%. Silica gel thin-layer chromatography (TLC) (solvent system CHCl3:MeOH 4:1) analysis of the MeOH fraction showed that one genotype had no phenolic compounds and two had only simple phenols. Once flavonol glycoside was present in relatively large amounts in four of the genotypes, but absent in genotypes with anthocyanins. Cellulose TLC (2-dimensional, Butanol:Acetic Acid:H2O 4:1:5 first dimension, 1% HCl second dimension) of the anthocyanin-containing genotypes showed that the presence of one flavonol and three anthocyanidin-3-glycosides (UV spot color and color shift with NH3). The relative importance of the seed coat chemicals in digestibility and their antioxidant will also be discussed.

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Clifford W. Beninger, George L. Hosfield and Mark J. Bassett

Three dry bean (Phaseolus vulgaris L.) genotypes differing in seedcoat color, mineral brown (P C D J G B v), yellow brown (P C D J G b v), and pale greenish yellow (P C D J g b v), were analyzed phytochemically. Kaempferol 3-O-β-d-glucoside (astragalin) was isolated and identified by nuclear magnetic resonance spectroscopy from all three genotypes, and was the main flavonoid monomer present. Flavonoid polymers (condensed tannins) were detected by thin layer chromatography, but anthocyanins were not detected in the three genotypes. High pressure liquid chromatography analyses indicated that astragalin was present at similar concentrations in pale greenish yellow and mineral brown genotypes, but was significantly lower in yellow brown. Presently, we do not know the functions of the G and B color genes, although the presence of astragalin in the three genotypes studied indicates these genes do not appear to act in a qualitative manner with regard to astragalin production, but may control the amount of astragalin present. Subtle differences in color between these genotypes may be due to the amount and type of tannins which have secondarily polymerized with phenolics and flavonoid monomers.

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George L. Hosfield, James D. Kelly and Mark A. Uebersax

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Clifford W. Beninger, George L. Hosfield, Mark J. Bassett and Shirley Owens

Three common bean (Phaseolus vulgaris L.) seedcoat color (or glossiness) genotypes, differing from each other by a single substitution at a seedcoat locus, were analyzed for presence and concentration of three anthocyanins: delphinidin 3-O-glucoside, petunidin 3-O-glucoside, and malvidin 3-O-glucoside. The three anthocyanins were present in Florida common bean breeding line 5-593 (P C J G B V Asp), matte black (P C J G B V asp), and dark brown violet (P C J G b V Asp), but the amounts varied greatly depending on the genotype. Dark brown violet had 19% of the total anthocyanin content when compared to 5-593, whereas matte black had amounts intermediate between the two other genotypes. The B gene acts to regulate the production of precursors of anthocyanins in the seedcoat color pathway above the level of dihydrokaempferol formation, perhaps at the chalcone synthase or chalcone isomerase steps in the biosynthetic pathway. We hypothesize that B regulates simultaneously the flavonoid (color) and isoflavonoid (resistance) pathways. The I gene for resistance to bean common mosaic virus (BCMV) is known to be linked closely to B. It is therefore hypothesized that the I gene function may be to respond to BCMV infection by dramatically increasing (over a low constituitive level) production in the 5-dehydroxy isoflavonoid pathway, which leads to synthesis of the major phytoalexin, phaseollin, for resistance to BCMV. Alternatively, the B and I genes may be allelic. The Asp gene affects seedcoat glossiness by means of a structural change to the seedcoat. We demonstrate that Asp in the recessive condition (asp/asp) changes the size and shape of the palisade cells of the seedcoat epidermis, making them significantly smaller than either 5-593 or dark brown violet. Asp, therefore, limits the amounts of anthocyanins in the seedcoat by reducing the size of palisade cells.

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Kimberly J. Walters, George L. Hosfield, Mark A. Uebersax and James D. Kelly

Three populations of navy bean (Phaseolus vulgaris L.), consisting of recombinant inbred lines, were grown at two locations for 2 years and were used to study canning quality. The traits measured included visual appeal (VIS), texture (TXT), and washed drained mass (WDM). Genotype mean squares were significant for all three traits across populations, although location and year mean squares were higher. We found a positive correlation (r = 0.19 to 0.66) between VIS and TXT and a negative correlation (r = -0.26 to -0.66) between VIS and WDM and between TXT and WDM (r = -0.53 to -0.83) in all three populations. Heritability estimates were calculated for VIS, TXT, and WDM, and these values were moderate to high (0.48 to 0.78). Random amplified polymorphic DNA markers associated with quantitative trait loci (QTL) for the same canning quality traits were identified and studied in each population. Marker-QTL associations were established using the general linear models procedure with significance set at P=0.05. Location and population specificity was common among the marker-QTL associations identified. Coefficient of determination (R2) values for groups of markers used in multiple regression analyses ranged from 0.2 to 0.52 for VIS, 0.11 to 0.38 for TXT, and 0.25 to 0.38 for WDM. Markers were identified that were associated with multiple traits and those associations supported correlations between phenotypic traits. MAS would offer no advantage over phenotypic selection for the improvement of negatively associated traits.

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Frank M. Elia, George L. Hosfield, James D. Kelly and Mark A. Uebersax

A knowledge of the relative proportion of additive and nonadditive genetic variances for complex traits in a population forms a basis for studying trait inheritance and can be used as a tool in plant breeding. A North Carolina Design II mating scheme was used to determine the inheritance of cooking time, protein and tannin content, and water absorption among 16 genotypes of dry bean (Phaseolus vulgaris L.) representative of the Andean Center of Domestication. Heritability and the degree of dominance for the traits were also calculated to provide guidelines for adopting breeding strategies for cultivar development. Thirty-two progeny resulted from the matings and these were assigned to two sets of 16 progeny each. Variances due to general combining ability (GCA) and specific combining ability (SCA) were significant for the traits. The GCA was larger in all cases. Narrow-sense heritability for protein, tannin, water absorption, and cooking time averaged 0.88, 0.91, 0.77, and 0.90, respectively. Degree of dominance estimates indicted that the traits were governed by genes with partial dominance except, in one case, tannin had a degree of dominance value of zero, indicating no dominance. The phenotypic correlation (-0.82) between water absorption and cooking time justifies using the water absorption trait as an indirect selection method for cooking time. With regard to parent selection in crosses, significant differences between GCA females and GCA males suggested cytoplasmic influences on trait expression. Hence, the way a parent is used in a cross (i.e., as female or male) will offset trait segregation. Using fast-cooking bean cultivars in conjunction with fuel-efficient cooking methods may be the best strategy to conserve fuelwood and help reduce the rate of deforestation in East and Central Africa.

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Maria-Carmela T. Posa-Macalincag, George L. Hosfield, Kenneth F. Grafton, Mark A. Uebersax and James D. Kelly

Canning quality of dry bean (Phaseolus vulgaris L.), of which the degree of splitting (SPLT) and overall appearance (APP) of canned beans are major components, is a complex trait that exhibits quantitative inheritance. The objectives of this study were to identify major genes that affect APP and SPLT in kidney bean, and map the location of these loci to the integrated core map of common bean. The analysis was performed using random amplified polymorphic DNA (RAPD) markers and two populations of kidney bean, consisting of 75 and 73 recombinant inbred lines (RILs), respectively. The two populations—`Montcalm' × `California Dark Red Kidney 82' and `Montcalm' × `California Early Light Red Kidney'—were planted in six year-location combinations in Michigan, Minnesota and North Dakota from 1996 to 1999. Correlations between APP and SPLT were high (0.91 to 0.97). Heritability estimates for APP and SPLT ranged from 0.83 to 0.85 in the two populations. Major genes for these traits were identified on two linkage groups. The first QTL, associated with seven RAPD markers, was putatively mapped to the B8 linkage group of the core bean linkage map. Desirable canning quality appeared to be derived from Montcalm at this locus. The second QTL, associated with four markers, appeared to be derived from the California parents. The second linkage group was not assigned to a linkage group in the core map. Population and environment-specificity were observed for the markers identified.