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Haytham Z. Zaiter, Dermot P. Coyne, James R. Steadman, and James S. Beaver

Genetic variation for abaxial leaf pubescence was detected among dry bean (Phaseolus vulgaris L.) cultivars/lines. Inheritance of pubescence (long, straight hairs) was studied in the dry bean crosses of pubescent `Pompadour Checa-50' (Dominican Republic) × eight glabrous cultivars/lines. Segregation for pubescence vs. glabrousness indicated that pubescence was determined by a single major gene or by duplicate recessive epistatic genes, depending on the cross involved. Trichome density (number trichomes per mm) was a quantitative trait. Thus, pubescence was a discrete trait, but trichome density ranged from low to high.

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Haytham Z. Zaiter, Dermot P. Coyne, Ralph B. Clark, and James R. Steadman

Nine bean cultivars/lines (Phaseolus vulgaris L.) were grown in three soils/rooting media at pH values of 7.9, 6.5, and 5.8 in greenhouse, growth chamber, and field experiments to evaluate the leaf reaction of the plants to a Nebraska bean rust [Uromyces appendiculatus (Pers.) Unger var. appendiculatus] isolate US85-NP-10-1. Significant differences were observed for rust pustule diameter between cultivars/lines grown in the three growth media. Plants grown in the medium at pH 5.8 showed significantly larger rust pustule diameters than those of plants grown at pH 6.5 or 7.9. A significant interaction occurred between growth medium and cultivars/lines for the rust reaction. Concentrations of Cl and Mn in leaves were positively correlated with rust pustule diameter. In contrast, concentration of K in leaves was negatively correlated with rust pustule diameter. Plant breeders attempting to improve beans for rust resistance must consider the growth medium pH in evaluating intensity and severity of rust symptoms on leaves.

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Soon O. Park, Dermot P. Coyne, Nedim Mutlu, Geunhwa Jung, and James R. Steadman

Common bacterial blight, incited by Xanthomonas campestris pv. phaseoli (Xcp) is a serious disease of common bean (Phaseolus vulgaris L.). Randomly amplified polymorphic DNA (RAPD) markers and flower color (V gene) previously were reported to be associated with six quantitative trait loci (QTL) affecting leaf and pod resistance to Xcp. However, the markers for the QTL were not confirmed in different populations and environments to indicate their merit in breeding. The objective was to determine if the associations of RAPD markers and the V gene with QTL for leaf and pod resistance to Xcp in a recombinant inbred (RI) backcross population from the cross BC2F6 `PC-50' × XAN-159 and for leaf resistance to Xcp in an F2 population from a different cross pinto `Chase' × XAN-159 could be confirmed. One or two genes from XAN-159 controlled leaf and pod resistance to Xcp. Among six QTL previously detected, five in the RI backcross population and three in the F2 population were confirmed to be associated with resistance to Xcp. The V gene and RAPD marker BC437.1050 on linkage group 5 were most consistently associated with leaf and pod resistance to two to five Xcp strains in the RI backcross population and with leaf resistance to two Xcp strains in the F2 population. One to three QTL affecting leaf and pod resistance to Xcp accounted for 22% to 61% of the phenotypic variation. Gene number (one to two) estimations and number of QTL (one to three) detected for leaf and pod resistance to Xcp in the RI backcross population were generally in agreement. The marker BC437.1050 and V gene, along with other resistance genes from other germplasm, could be utilized to pyramid the different genes into a susceptible or partially resistant bean line or cultivar to enhance the level of resistance to Xcp.

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Tae-Jin Lee, Dermot P. Coyne, Thomas E. Clemente, and Amitava Mitra

Expression of lactoferrin (LF) gene, a cationic iron-binding glycoprotein, was investigated in transgenic tomato plants (Lycopersicon esculentum Mill.). Resistance of the transgenic tomato plants to the pathogen (Ralstonia solanacearum Smith) causing bacterial wilt was also determined. Tomato line F7926-96, susceptible to bacterial wilt, was transformed with Agrobacterium strain C58C1 containing a plasmid construction carrying a modified LF cDNA. The introgression of LF cDNA into the susceptible tomato line was confirmed by Southern blot and the expression of full-length lactoferrin transcript and protein was also detected by northern and western blots, respectively. Based on resistance to kanamycin, a Mendelian segregation for a single locus insertion was observed in the T1 and T2 generations and all T1 and T2 plants resistant to kanamycin showed the single corresponding band of LF cDNA in Southern blot analysis. Two transgenic tomato lines inoculated with 1 × 107 and 1 × 108 colony-forming units (CFU)/mL with Rs isolate NC251 (K60, race 1) exhibited early resistance and subsequent susceptibility, while 44% to 55% of plants survived until maturity (fruit ripening) when inoculated with 1 × 105 CFU/mL in comparison with the fully susceptible tomato line. The latter resistance to bacterial wilt in transgenic tomatoes with the stable Mendelian segregation patterns for the LF gene suggests a potential new approach to consider for control of bacterial wilt of tomato. The possible value of this gene along with other plant genes to control bacterial pathogens is discussed.

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Muharrem Ergun, Ellen T. Paparozzi, Dermot P. Coyne, Durward Smith, Stephen Kachman, and David S. Nuland

Seedcoat color is an important trait, as it affects marketing and consumer acceptance of pinto beans (Phaseolus vulgaris L.). Pinto breeding line NE 94-4 showed seedcoat yellowing in on-farm field trials in Nebraska in 1996 and 1997. Hail, sprinkler irrigation, and fall rainfall appeared to be involved in increasing seedcoat yellowing, based on analysis of field and weather data of on-farm trial sites. The objective of this study was to determine the effect of moisture on seedcoat yellowing of pinto line NE 94-4 (susceptible) and pinto `UI-114' (highly resistant). Two greenhouse experiments were conducted involving misting of bean plants near maturity and injecting water into maturing bean pods. Another experiment evaluated the response of seeds of these two bean entries to moisture by placing them on moist filter paper in petri dishes in the laboratory. Results showed that both genotype and moisture content are involved in seedcoat yellowing. This simple, cheap, and effective filter paper test was then used to evaluate seedcoat yellowing of nine pinto genotypes in response to moisture. Pinto NE 94-4 and `Kodiak' showed the greatest change, while `Bill Z' showed the least change, in seedcoat color.

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Dermot P. Coyne, James M. Reiser, Durward Smith, Lisa Sutton, Aly M. Ibrahim, and Dale Lindgren

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Geunhwa Jung, Dermot P. Coyne, James Bokosi, James R. Steadman, and James Nienhuis

Dry bean (Phaseolus vulgaris L.) production is limited by bean rust [Uromyces appendiculatus (Pers.) Unger var. appendiculatus]. An effective control strategy for this disease is to breed cultivars with durable resistance. Information on the inheritance, genetic relationships, and mapping of genes with molecular markers for specific resistance (SR), adult plant resistance (APR), and abaxial leaf pubescence (ALP) is needed to pyramid the desired genes for durable resistance. ALP was found to be associated previously with APR in Andean germplasm. The objective here was to identify and map RAPD markers for the genes controlling SR, APR, and ALP and to examine their relationships. Five rust pathotypes were inoculated on the unifoliate leaves of 68 recombinant inbred (RI) lines derived from `PC-50' (presence of SR, APR, and ALP) × XAN-159 (absence of SR, APR, and ALP). SR was determined by a single major gene (Ur-9) to the five rust pathotypes with no detection of recombinants. The fourth trifoliolate leaves were inoculated with one pathotype (A88T1-4b). A single major gene Ur-12 controlled APR to that pathotype. The Ur-9 gene (SR) was independent of and epistatic to the Ur-12 gene (APR). Because of the low number of APR lines in the RI population resulting from the elimination of RI lines with SR, an F2 population was developed from a cross of two homozygous RI lines selected for unifoliate susceptibility to pathotype A88T1-4b and for resistance and susceptibility of the fourth trifoliolate leaves to tag RAPD markers linked to the Ur-12 gene (APR). The single major gene Pu-a determinated ALP and was not linked to Ur-9 (SR) and Ur-12 (ALP). The gene Ur-9 (SR) was linked to RAPD marker J13-1100 at 5 cM and was not assigned to any linkage group or other markers. The gene Pu-a (ALP) was mapped at 20.2 cM from 116.500 and 3.9 cM from marker G3.1150 in linkage group 3. The Ur-12 gene (APR) was mapped at 34.6 cM from marker O13.1350 in linkage group 4b. This is the first report of mapping a gene for APR in common bean.

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Soon O. Park, Dermot P. Coyne, James R. Steadman, and Geunhwa Jung

White mold, incited by Sclerotinia sclerotiorum (Ss), is an important disease of common bean (Phaseolus vulgaris). Our objective was to identify RAPD markers and seedcoat pattern associated with QTL affecting resistance to Ss isolates 152 and 279 in a molecular marker-based linkage map previously constructed using a recombinant inbred (RI) population from the common bean cross `PC-50' (resistant to Ss) x XAN-159 (susceptible to Ss). White mold reactions were derived from a greenhouse straw test. Continuous distributions for the reactions to Ss isolates 152 and 279 were observed for RI lines, indicating quantitative inheritance. An intermediate (+0.67) Pearson correlation was observed between the reactions to Ss isolates 152 and 279. Low (0.24 and 0.23) narrow-sense heritabilities were found for the reactions to Ss isolates 152 and 279. Three QTL affecting resistance to Ss isolate 152 explained 33% of the phenotypic variation. Four QTL affecting resistance to Ss isolate 279 explained 54% of the phenotypic variation. The seedcoat pattern marker (C) on linkage group I was most consistently associated with resistance to Ss isolates 152 and 279, and explained 10% and 24% of the phenotypic variation for the traits, respectively. This is the first report on detection of QTL for white mold resistance in common bean. The RAPD markers and seedcoat pattern could be useful in breeding for white mold resistance.

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Geunhwa Jung, Paul W. Skroch, Dermot P. Coyne, James Nienhuis, and E. Arnaud-Santana

Common bacterial blight (CBB) incited by the bacterial pathogen Xanthomonas campestris pv. phaseoli (Smith) Dye is an important disease of common bean. In a previous study, QTL associated with CBB resistance were described based on RAPD marker analysis of a recombinant inbred population derived from the common bean cross BAC-6 (R) × HT-7719 (S) (resistant × susceptible). The objective of this research is to confirm these previously described candidate marker locus-QTL associations using an inbred backcross PC-50 (S) × BAC-6 (R) and a recombinant inbred Venezuela 44 (S) × BAC-6 (R) population. Two markers previously found to be associated with QTL for CBB resistance in the BAC-6 × HT-7719 population were found to account for 30% of the phenotypic variation for CBB resistance in the PC-50 × BAC-6 inbred backcross population. The three most resistant BC2F3 lines based on marker locus genotypes were ranked 1, 3, and 7 (out of 64) based on phenotypic evaluation. These results provide important confirmation of marker locus-QTL associations and indicate that RAPD markers linked to loci controlling the expression of CBB resistance in common bean may be used to transfer resistance genes into susceptible breeding material.

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Soon O. Park, Dermot P. Coyne, Nedim Mutlu, James R. Steadman, and Geunhwa Jung

Common bacterial blight, incited by Xanthomonas campestris pv. phaseoli (Xcp), is a serious disease of common bean (Phaseolus vulgaris). RAPD markers and flower color (V gene) previously had been reported to be associated with six QTL affecting leaf and pod resistance to Xcp. However, the markers for the QTL were not confirmed in different populations and environments to indicate their merit in breeding. Our objective was to determine if the associations of RAPD markers and the V gene with QTL for leaf and pod resistance to Xcp in a RI backcross population from the cross BC2F6 `PC-50' × XAN-159 and for leaf resistance to Xcp in a F2 population from a different cross Pinto `Chase' × XAN-159 could be confirmed. Among six QTL previously detected, five in the RI backcross population and three in the F2 population were confirmed to be associated with resistance to Xcp. The V gene and RAPD marker BC437.1050 on linkage group 5 were most consistently associated with leaf and pod resistance to two to five XCP strains in the RI backcross population and with leaf resistance to two Xcp strains in the F2 population. The confirmed marker BC437.1050 and V gene on linkage group 5, along with other resistance genes from other germplasm, could be used to pyramid the different genes into a bean cultivar to enhance the resistance to Xcp.