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James R. McFerson, Warren F. Lamboy, and Steve Kresovich

57 WORKSHOP 8 Use of Molecular Markers in Germplasm Management

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

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

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Thomas Horejsi and Jack Staub

106 POSTER SESSION (Abstr. 335–343) Breeding and Genetics–Vegetables II (Molecular Markers and Physiological Genetics)

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S.O. Park, A. Dursun, D.P. Coyne, and G. Jung

106 POSTER SESSION (Abstr. 335–343) Breeding and Genetics–Vegetables II (Molecular Markers and Physiological Genetics)

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Chunxiao Jiang and Kenneth C. Sink

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

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Winthrop B. Phippen, James R. McFerson, and Stephen Kresovich

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

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Mario Crespo, James Nienhuis, Jan Tivang, and Paul Skroch

Knowledge of relative genetic distance among genotypes is useful in a breeding program because it permits organization of germplasm resources. Genetic distance (GD) was estimated among 113 faba bean, Vicia faba L. genotypes, which included three botanical varieties from different geographical areas around the world. The genotypes included 87 accessions from Bolivia, 14 accessions from the Middle East and North Africa, five accessions from Australia, and seven commercial varieties from Europe. Twenty-three RAPD primers were scored yielding four to 13 polymorphic bands resulting in a total of 165 bands. Our objective was to determine genetic relationships among accessions and cultivars as measured by RAPD markers. The genetic relationships were estimated using the ratio of discordant to total bands scored. A multidimensional scaling (MDS) plot indicated four clusters corresponding to: i) European commercial cultivars; ii) the Middle East, North Africa, and Australian accessions; iii) the Bolivian highland landraces; and iv) the Bolivian collection maintained in a valley environment. A permutation test confirmed the four clusters (P < 0.01). Sampling variance results indicated that a CV of 10% could be obtained with as few as 148 bands between groups. Selection and drift appears the main cause of divergence of two populations in the Bolivian faba bean collection. The results of this study indicated that RAPDs are a powerful tool for evaluation of germplasm conservation methods in faba bean.

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M.R. Stevens, P.D. Griffiths, J.W. Scott, D.K. Heiny, and D.D Rhoads

Sw-5 is a locus introgressed from Lycopersicon peruvianum to some L. esculentum lines conferring dominant resistance to TSWV. Restriction fragment length polymorphism (RFLP) analyses positions Sw-5 to the long arm of chromosome 9 in the sub-telomeric region between CT71 and CT220. RFLP analyses suggest the introgressed region begins distal to CT71, includes CT220, and may extend to the telomere. Randomly amplified polymorphic DNA (RAPD) analyses with >700 random 10-mer primers identified a single 2.2-kbp band with one primer (primer #72 GAGCACGGGA) that is tightly linked to Sw-5. However, we have also produced an equivalent 2.2-kbp band in analysis of other TSWV-susceptible tomato breeding lines. Thus, this band likely derives from L. esculentum DNA very near to Sw-5 and the introgressed region. Additional analyses have recently detected a potential co-dominant RAPD polymorphism linked to Sw-5.

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James M. Bradeen and Philipp Simon

The Y2 locus conditions α- and β-carotene accumulation in the xylem (core) of carrot roots. The dominant allele suppresses carotene, but not xanthophyll accumulation, resulting in yellow-cored roots. Individuals homozygous for the recessive allele are rich in carotenes and are therefore orange-cored. Increased consumer interest in high carotene produce requires improved understanding of carotene biosynthesis and color development and more-efficient breeding techniques. We examined 103 F2 individuals generated from inbred populations differing in core carotene content. Bulked segregant analysis identified AFLP bands putatively linked to Y2. Linkage was confirmed for some bands by mapping. Linked bands were excised from gels, re-amplified, cloned into pGEM vectors, and sequenced. Cloned fragments and sequence information were used to characterize larger genomic regions to identify codominant markers. Currently we are developing codominant, PCR-based markers that can be used to rapidly genotype individuals in breeding programs, to characterize wild, feral, and cultivated populations for diversity and evolution studies, and to examine the role of Y2 in carotene accumulation.

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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.