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Douglas G. Bielenberg, Ying Wang, Gregory L. Reighard and Albert G. Abbott

`Evergrowing' (evg) peach is a naturally occurring mutant unable to enter winter dormancy in response to dormancy inducing conditions. The evg mutant is one of only two described mutants affecting winter dormancy in woody perennial trees. The evg mutation segregates as a single recessive gene and previous work by our group has fine mapped the trait between flanking markers separated by 3.3 centiMorgans. This region was physically mapped using a bacterial artificial chromosome (BAC) library from and a contig of overlapping genomic fragments identified. We have utilized several approaches to complete the sequencing of a 132 kilobase region of the peach genome derived from three overlapping BACs that encompass the complete EVG gene containing region. We present here our analysis and annotation of the genomic region, including putative and experimentally verified gene coding sequences. A primary feature of the region is a large tandem duplication of a region containing a MADS-box type DNA binding transcription factor resulting in six similar copies of the gene, all of which appear to be expressed at the mRNA level in wild-type germplasm. Hybridization analysis revealed the presence of a large deletion in the mutant genome. Five of the identified genes fall within the evg mutation and represent new candidates for the control of entrance into winter dormancy.

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Ying Wang, Gregory L. Reighard, Desmond R. Layne, Albert G. Abbott and Hongwen Huang

Pawpaw (Asimina triloba) produces the largest fruit native to the United States. Six linkage groups were identified for A. triloba using the interspecific cross [PPF1-5 (A. triloba) × RET (A. reticulata Shuttlw. ex Chapman)], covering 206 centimorgans (cM). A total of 134 dominant amplification fragment length polymorphism (AFLP) markers (37 polymorphic and 97 monomorphic) were employed for estimating the genetic diversity of eight wild populations and 31 cultivars and advanced selections. For the wild populations, the percentage of polymorphic loci over all populations was 28.1% for dominant markers and Nei's genetic diversity (He) were 0.077 estimated by 134 dominant markers. Genetic diversity and the percentage of polymorphic loci estimated using only polymorphic dominant AFLPs were 0.245 and 79%, respectively, which are comparable with other plant species having the same characteristics. Estimated genetic diversity within populations accounted for 81.3% of the total genetic diversity. For cultivars and advanced selections, genetic diversity estimated by 134 dominant markers was similar to that of wild pawpaw populations (He = 0.071). Thirty-one cultivars and advanced selections were delineated by as few as nine polymorphic AFLP dominant loci. Genetic relationships among wild populations, cultivars and advanced selections were further examined by unweighted pair group method with arithmetic mean (UPGMA) of Nei's unbiased genetic distance. The genetic diversity estimated for wild populations using the clustered polymorphic markers was lower than the result estimated using the nonclustered polymorphic markers. Therefore, this study indicates that the number of sampled genomic regions, instead of the number of markers, plays an important role for the genetic diversity estimates.

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Ying Wang, Laura L. George, Gregory L. Reighard, Ralph Scorza and Albert G. Abbott

Evergreen genotypes of peach [Prunus persica (L.) Batsch] have been identified in Mexico, where terminal growth on evergreen trees is continuous under favorable environmental conditions. This evergreen trait in peach is controlled by one single gene (evg), and this evergreen condition is homozygous recessive. Four dominant AFLP markers, EAT/MCAC, ETT/MCCA2, EAT/MCTA, and ETT/MACC, were found to be tightly linked to the evg locus at 1 cM, 4.6 cM, 5.8 cM, and 11 cM, respectively. All four markers were sequenced and identified. A peach BAC library was constructed by using the pBeloBAC11 vector for building the physical map for the evg gene. This library represents four times the coverage of the peach genome with the average insert size of 50 to 70 kb. The EAT/MCAC AFLP marker fragment was used for screening the peach BAC library. A single BAC clone, 18F12, was confirmed to contain this fragment. The final BAC contig for this evg gene region and the potential homology between peach and Arabidopsis thaliana will be presented and discussed.