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Mark E. Lewis and Fred A. Bliss

Ten common bean (Phaseolus vulgaris L.) lines—including cultivars, breeding lines, and one wild line—were evaluated for susceptibility to Agrobacterium tumefaciens strain C58 by stab-inoculating intact shoot tips of germinating seeds. Significant differences for tumor frequency and size were found on the resulting 3-week-old seedlings. UW 325, a wild bean, had the highest rate of tumorigenesis; `Olathe', a dry bean cultivar, had the lowest. Uninoculated excised shoot tips cultured in media with BA or BA plus NAA exhibited differences in phytohormone sensitivity, as evidenced by callusing and root initiation. The cultivar Montcalm seemed to be highly sensitive, while `Olathe' was relatively insensitive. Fluorometric GUS assays of shoot tips from germinating seeds inoculated with the disarmed GUS-containing A. tumefaciens strain C58C1(pGV3850/pKIWI105) showed that UW 325 had the highest level of GUS activity. `Montcalm' had a high rate of tumorigenesis but a low level of GUS activity; this anomaly was attributed to its high phytohormone sensitivity. The use of the virulence-inducing compound acetosyringone in the inoculum culture medium did not alter genotypic differences (ranks) in susceptibility. Histochemical GUS assays of inoculated UW 325 shoot tips showed that 60% of the apexes exhibited one or more transformation events. Chemical names used: β-glucuronidase (GUS); α-naphthaleneacetic acid (NAA); N-(phenylmethyl)-1H-purin-6-amine (BA).

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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.

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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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Agnes A. Flores-Nimedez, Paul H. Li, and Charles C. Shin

GLK-8903, an experimental product whose main ingredient is produced by hydrogenation of a primary alcohol extracted from plants, showed significant potential in protecting bean (Phaseolus vulgaris L.) plants from chilling injury. The GLK-8903 protection mechanism was assessed by examining several physiological and biochemical responses. The decline in leaf water potential and the increase in osmotic potential caused by chilling exposure to 4C (day/night) were minimized by the application of GLK-8903. Chilling causes an increase in electrolyte leakage, an indication of chilling injury of the plasma membrane. Increased electrolyte leakage was reduced significantly in the GLK-8903-treated plants during chilling. This minimized leakage may be due to less damage of the plasma membrane. Plasmolysis and deplasmolysis studies of the epidermal cells suggest that GLK-8903 is able to reduce the plasma membrane perturbation in the chilling environment, as evident by: 1) the lower permeability coefficient to urea at 4C, and 2) the swelling of protoplasts in the cells of untreated tissues after chilling exposure with no swelling of the protoplast being observed in the GLK-8903-treated cells. Malondialdehyde (MDA), a product of lipid peroxidation, increased more in untreated controls than in treated plants exposed to 4C. Plasma membrane ATPase activity decreased less in GLK-8903-treated plants than in untreated controls after 3 days at 4C. The mechanism of GLK-8903-alleviated chilling injury is discussed.

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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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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, 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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Antoni Almirall, Lluís Bosch, Roser Romero del Castillo, Ana Rivera, and Francesc Casañas

foodstuffs. European Commission. 2006b. Regulation (CE) 509/2006, on the guaranteed traditional specialities for agricultural products and foodstuffs. IBPGR 1982 Phaseolus vulgaris descriptors IBPGR Secretariat Rome, Italy

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Sameh Sassi-Aydi, Samir Aydi, and Chedly Abdelly

). Several authors have addressed the inhibitory effect of osmotic stress on root nodule activity in legumes ( King and Purcell, 2001 , 2005 ; Marino et al., 2006 ; Serraj and Sinclair, 1997 ) including common bean ( Phaseolus vulgaris L.) ( Sassi et al

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Rie Sadohara, James D. Kelly, and Karen A. Cichy

Common beans ( Phaseolus vulgaris L.) are a good source of protein, dietary fiber, vitamins, and minerals and offer numerous health benefits to consumers ( Hayat et al., 2014 ; Messina, 2014 ). However, the annual per capita consumption of beans