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

Bean rust, caused by Uromyces appendiculatus, is a major disease of common bean (Phaseolus vulgaris). The objective was to identify RAPD markers linked to the gene (Ur-7) for specific resistance to rust race 59 using bulked segregant analysis in an F2 segregating population from the common bean cross GN1140 (resistant to rust) × Nebraska #1 (susceptible to rust). A single dominant gene controlling specific resistance to race 59 was found in the F2 and was confirmed in the F3. Seven RAPD markers were detected in a coupling-phase linkage with the Ur-7 gene. Coupling-phase RAPD markers OAA11.500, OAD12.550, and OAF17.900 with no recombination to the Ur-7 gene were found. Three RAPD markers were identified in a repulsion-phase linkage with the Ur-7 gene among the three markers at a distance of 8.2 cM. This is the first report on RAPD markers linked to the Ur-7 gene in common bean. The RAPD markers linked to the gene for specific rust resistance of Middle American origin detected here, along with other independent rust resistance genes from other germplasm, could be used to pyramid multiple genes into a bean cultivar for more-durable rust resistance.

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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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Yiping Zhang and John R. Stommel

The carotenoids have an important influence on tomato fruit quality and enhance the fruit contribution to human nutrition. Expression of the high pigment (hp) locus in tomato results in increased total carotenoids and increased efficiency of utilization of the polyenes. A similar mutant, dark green (dg), contains higher level of chlorophyll in immature fruit and results in darker red pigmentation, both externally and internally in ripe fruit. Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analyses were performed using two pairs of near isogenic lines (NILs) designed to be isogenic at the hp and dg loci. Sixty-four AFLP primer pairs and more than 1000 RAPD 10-mer primers were screened for polymorphism between each pair of the NILs. One RAPD marker was identified to be linked to the hp gene, and two AFLP primer pairs showed polymorphic fragments which distinguished the dg NILs. The markers identified in this study will be converted to allele specific SCAR (sequence characterized amplified region) markers, which are more useful in marker-assisted selection breeding programs.

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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.

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Z. Wiesman, N. Avidan, S. Lavee, and B. Quebedeaux

RAPD analysis was performed on the main traditional and introduced varieties of olive grown in Israel and the West Bank. Primers were identified and used in combination to discriminate between different varieties. Significant biodiversity was demonstrated among `Nabali' olive trees growing along the central mountain ridge of the West Bank, suggesting that the grouping known as `Nabali' is actually a mixture of genetically distinct variants. On the other hand, RAPD profiles of selected variants of `Souri', cultivated mainly in the northern mountains of Israel, revealed a high degree of similarity, indicating that these variants represent environmental phenotypes of the same genome. Molecular differences were demonstrated between the `Nabali' group variants and `Souri'. Other more recently developed or introduced varieties showed individually distinct RAPD profiles. Possible future developments based on these data are mentioned.

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Geunhwa Jung, Dermot P. Coyne, Paul W. Skroch, James Nienhuis, E. Arnaud-Santana, James Bokosi, H.M. Ariyarathne, James R. Steadman, James S. Beaver, and Shawn M. Kaeppler

Random amplified polymorphic DNA (RAPD) markers were used to construct a partial linkage map in a recombinant inbred population derived from the common bean (Phaseolus vulgaris L.) cross BAC 6 × HT 7719 for studying the genetics of disease resistance in common bean. The linkage map spanned 545 cM and included 75 of 84 markers used in this study. The population of 128 recombinant inbred lines was evaluated for resistance to common bacterial blight, foliar resistance to web blight [WB; Thanatephorus cucumeris (Frank) Donk], and resistance to rust [Uromyces appendiculatus var. appendiculatus (Pers.:Pers) Unger]. Common bacterial blight [CBB; Xanthomonas campestris pv. phaseoli (Smith) Dye] resistance was evaluated for CBB strain Epif-IV in later-developed trifoliolate leaves and for CBB strain EK-11 in seeds, first trifoliolate leaves, later-developed trifoliolate leaves, and pods. In addition, lines were rated for plant uprightness and branch density. Two to six markers accounted for 14% to 34% of the phenotypic variation for each trait. Significant marker locustrait associations were found for 14 mapped loci and 7 of the 9 unmapped markers. The distribution of detected QTL appeared to be nonrandom with most significant markers associated with more than one trait or closely linked to markers significantly associated with variation for a different trait. One marker, BC4091250, was significantly associated with WB resistance, resistance for CBB strain Epif-IV in later-developed trifoliolate leaves, and resistance for CBB strain EK-11 in first trifoliolate leaves, later-developed trifoliolate leaves, and pods. A rust resistance gene was mapped in an interval 14.6 cM from RAPD marker H191050 and 12.5 cM from marker AJ16250.

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Shinya Kanzaki, Keizo Yonemori, Akira Sugiura, Akihiko Sato, and Masahiko Yamada

Japanese persimmon (Diospyros kaki Thunb.) cultivars are classified into four types depending upon the nature of astringency loss of the fruit. Among them, the pollination-constant and nonastringent (PCNA) type is the most desirable for fresh fruit consumption due to the trait of stable loss of astringency on the tree with fruit development. Lack of tannin accumulation is the main cause of natural astringency loss in PCNA-type fruit, and is qualitatively inherited. The PCNA trait is recessive to the non-PCNA trait. In this study, we investigated amplified fragment length polymorphism (AFLP) markers for the trait of natural astringency loss of PCNA-type fruit using bulked segregant analysis (BSA) for efficient selection of PCNA type plants in a breeding population. A total of 128 primer combinations were tested and one AFLP marker was found to be linked to the dominant allele controlling the trait for astringency. This marker, EACC/MCTA-400, was absent in all of the PCNA-type plants tested, whereas it was present in about half of the non-PCNA-type plants tested. However, RFLP analysis using this marker enabled the detection of the other dominant allele, and all PCNA-type plants could be distinguished from the non-PCNA-type plants. Application of this marker system will be useful for the selection of PCNA-type plants in persimmon breeding.

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Courtney A. Weber, Gloria A. Moore, Zhanao Deng, and Fred G. Gmitter Jr.

Mapping quantitative trait loci (QTL) associated with freeze tolerance was accomplished using a Citrus grandis (L.) Osb. × Poncirus trifoliata (L.) Raf. F1 pseudo-testcross population. A progeny population of 442 plants was acclimated and exposed to temperatures of -9 °C and -15 °C in two separate freeze tests. A subpopulation of 99 progeny was genotyped for random amplified polymorphic DNA (RAPD), cleaved amplified polymorphic sequence (CAPS), sequence characterized amplified region (SCAR), and sequence tagged site (STS) markers to produce a linkage map for each parent. Potential QTL were identified by interval mapping, and their validity was corroborated with results from means comparison (t test), one-way analysis of variance (F test), and bulked segregant analysis (BSA). Multiple analytical methods provided evidence supporting putative QTL and decreased the probability of missing significant QTL associated with freeze tolerance. QTL with a large effect on freeze tolerance were located on both the Citrus and Poncirus linkage maps. In addition, clusters of markers with significantly different means between marker present and absent classes indicating minor QTL that contribute smaller effects on the level of tolerance were found on the linkage maps of both species.

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Charles J. Simon and Richard M. Hannan

30 POSTER SESSION 4 (Abstr. 460-484) Breeding/Genetics/Molecular Markers

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T. Casey Garvey and John D. Hewitt

An interspecific hybrid was made between an accession of Lycopersicon cheesmanii f. minor Riley (LA 1508) from the Galapagos Islands, Ecuador, and L. pennellii (Corr.) D'Arcy (LA 716). LA 1508 was used because of its high soluble solids content (SSC). It was crossed with LA 716 to test for linkage between isozymes and morphological markers and loci conditioning high SSC. For both accessions, chromosome numbers are equal and there are large differences between SSC and no barriers to crossing. Modified BC1 populations derived from the hybridization were assayed for isozyme markers using starch gel electrophoresis. Associations between marker loci and quantitative-trait loci (QTL) conditioning high SSC were determined using analysis of variance. Six isozymes located on five chromosomes and one morphological marker had significant associations with SSC, indicating linkage to QTL. Digenic epistatic interactions between pairs of independent markers did not appear to play an important role in the interactions between QTL that condition SSC.