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Fenny Dane, Leigh K. Hawkins, and Hongwen Huang

Genetic variation among nine populations of Ozark chinkapin [Castanea pumila (L.) Mill. var. ozarkensis (Ashe) Tucker], threatened by their susceptibility to chestnut blight (Cryphonectria parasitica (Murrill) Barr), was investigated. Population genetic parameters estimated from isozyme variation suggest the populations have a higher genetic diversity (He = 0.227) than populations of the other Castanea Mill. species on the North American continent, the American chestnut (C. dentata (Marsh.) Borkh.) High levels of heterozygosity were detected within the populations, but nonsignificant differences in genetic diversity were observed among the different populations. Principal component analysis based on isozyme allele frequencies or randomly amplified polymorphic DNA phenotype frequencies showed clustering of the same populations. Populations with high levels of genetic diversity and unusual alleles should be the focal point of conservation biologists for capturing much of the genetic variation of the species.

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Leigh K. Hawkins, Fenny Dane, and Thomas L. Kubisiak

Morphological traits were examined in an F3 generation derived from a cross between C. lanatus var. lanatus [(Thunb.) Matsum. & Nakai] and C. lanatus var. citroides. At least three genes, C (yellow) vs. c (red), i (inhibitory to C) vs. I (non-inhibitory to C), and y (yellow) vs. yw (white), with epistatic and inhibitory actions were found to govern the inheritance of fruit flesh color. The high frequency of yellow-fleshed fruit and low frequencies of white and red fruits can be explained by the presence of a new allele (yw recessive to y) in the multiple allele series at the Y locus. The low frequency of tan colored seeds in segregating populations could be explained by at least three genes governing inheritance of seed-coat color. Single factor analysis of variance was conducted for each pairwise combination of random amplified polymorphic DNA (RAPD) locus and fruit or seed characteristics. Several RAPD loci were identified to be loosely linked to morphological characteristics.

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Leigh K. Hawkins, Fenny Dane, Thomas L. Kubisiak, and Billy Rhodes

Fusarium wilt, caused by the soilborne fungus Fusarium oxysporum f.sp. niveum (FON), is a serious disease of the watermelon (Citrullus lanatus). Three races of this pathogen (races 0, 1, and 2) have been identified based on differential pathogenicity assays. Most commercially available cultivars are resistant to races 0 and 1. Inheritance for resistance to these races is thought to be controlled by a single dominant gene. No cultivars are resistant to race 2 and resistance is thought to be a quantitative trait. F2 lines derived from a cross between the Fusarium-resistant Citrullus lanatus PI296341, and the Fusarium-susceptible watermelon cultivar `New Hampshire Midget' were used to generate a RAPD-based map of the Citrullus genome. F2:3 families were assayed in the greenhouse for resistance to races 1 and 2. Those families that were either highly resistant or highly susceptible were used in identifying markers linked to Fusarium wilt resistance. A preliminary map of the Citrullus genome based on random amplified polymorphic DNA (RAPD) markers has been expanded with the inclusion of simple sequence repeats (SSRs), amplified fragment length polymorphisms (AFLPs), and isozymes.

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Leigh K. Hawkins, Fenny Dane, Tom Kubisiak, Bill Rhodes, and Bob Jarrett

A linkage map was constructed of the watermelon genome using F2 and F2:3 populations segregating for resistance to race 1 and 2 of Fusarium oxysporum f. sp. niveum (FON 1 and 2). Sixty-four percent of the RAPD primers used in the parents and F1 detected polymorphism. In the F2, 143 polymorphic bands were scored, 60% of which exhibited the expected 3:1 segregation ratio. A 113 cM linkage map was constructed using Mapmaker version 3 and LOD of 4. DNA pools of Fusarium wilt resistant or susceptible F2:3 lines were created and bulked segregant analysis was used to detect molecular markers linked to FON 1 or FON 2 resistance. Four individuals per line were used to confirm linkages and construct an F2:3 linkage map. One large linkage group was detected in both generations. A large proportion of the RAPD and SSR markers were unlinked and many showed segregation distortion. Single-factor ANOVA for each pairwise combination of marker locus and resistance or morphological trait was conducted. RAPD markers with putative linkages to FON 1 and FON 2 and several morphological traits were detected.

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Leigh K. Hawkins, Fenny Dane, Thomas L. Kubisiak, Billy B. Rhodes, and Robert L. Jarret

Isozyme, randomly amplified polymorphic DNA (RAPD), and simple sequence repeats (SSR) markers were used to generate a linkage map in an F2 and F3 watermelon [Citrullus lanatus (Thumb.) Matsum. & Nakai] population derived from a cross between the fusarium wilt (Fusarium oxysporum f. sp. niveum) susceptible `New Hampshire Midget' and resistant PI 296341-FR. A 112.9 cM RAPD-based map consisting of 26 markers spanning two linkage groups was generated with F2 data. With F3 data, a 139 cM RAPD-based map consisting of 13 markers covering five linkage groups was constructed. Isozyme and SSR markers were unlinked. About 40% to 48% of the RAPD markers were significantly skewed from expected Mendelian segregation ratios in both generations. Bulked segregant analysis and single-factor analysis of variance were employed to identify RAPD markers linked to fusarium wilt caused by races 1 and 2 of F. oxysporum f. sp. niveum. Current linkage estimates between the resistance trait and the marker loci were too large for effective use in a marker-assisted selection program.