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Kirk W. Pomper, Sheri B. Crabtree, Shawn P. Brown, Snake C. Jones, Tera M. Bonney, and Desmond R. Layne

The pawpaw [Asimina triloba (L.) Dunal.] is a tree fruit native to many areas of the southeastern and mid-western United States. Kentucky State University (KSU) is designated as a satellite repository for Asimina for the U.S. Department of Agriculture (USDA), National Plant Germplasm System (NPGS). An assessment of the level of genetic diversity in cultivated pawpaw would assist in development of the future germplasm repository collection strategies for cultivar improvement. The objectives of this study were to identify intersimple sequence repeat (ISSR) markers that segregate in a simple Mendelian fashion and to use these markers to assess genetic diversity in 19 pawpaw cultivars. Leaf samples from the 34 progeny of controlled crosses (1-7-1 × 2-54 and reciprocal) and the parents were collected, DNA was extracted, and subjected to the ISSR methodology using the University of British Columbia microsatellite primer set #9. Seven primers yielded 11 Mendelian markers with either a 3:1 or 1:1 ratio that was confirmed by chi-square analysis. Analysis of genetic diversity using 10 of the ISSR markers from 19 pawpaw cultivars revealed a moderate to high level of genetic diversity, with a percent polymorphic loci P = 80 and an expected heterozygosity He = 0.358. These diversity values are higher than those reported for cultivated pawpaw using isozyme or randomly amplified polymorphic DNA (RAPD) markers, indicating that the ISSR marker methodolgy has a higher level of discrimination in evaluating genetic diversity in pawpaw and/or pawpaw has greater levels of genetic diversity than previously found.

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James Nienhuis, Julie Rodriguez, Wilber Phillips, Peter Hanson, and Liliway Engle

Worldwide, there are cuurently more than 60 germplasm banks that contain tomato (Lycopersicon esculentum) collections ranging is size from a few dozen to several thousands of accessions. In the utilization of these genetic resources sampling from only one germplasm bank may result in limiting available genetic diversity, whereas sampling from several germplasm banks may result in unnecessary redundancy. The current lack of knowledge regarding the relative magnitudes of genetic diversity contained within different collections makes it difficult to develop a core collection that maximizes genetic diversity. Two large tomato collections are housed at the Asian Vegetable Research and Development Center (AVRDC), Sanhua, Taiwan, R.O.C., and the Centro Agronomico Tropical de Investigacion y Enseoanza (CATIE), Turrialba, Costa Rica. Ninety-six accessions from CATIE and 102 accessions from AVRDC were randomly sampled from each base collection. The total of 198 accessions were charcterized for 103 polymorphic RAPD molecular marker bands. The results indicated that the two germplam banks sampled different genetic diversity. In addition, the magnitude of genetic diversity was greater in the AVRDC collection compared to CATIE.

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Dilson A. Bisognin and David S. Douches

An understanding of the genetic relationship within potato germplasm is important to establish a broad genetic base for breeding purposes. The objective of this study was to assess the genetic diversity of potato (Solanum tuberosum subsp. tuberosum Hawkes) germplasm that can be used in the development of cultivars with resistance to late blight caused by Phytophthora infestans (Mont.) de Bary. Thirty-three diploid and 27 tetraploid late blight resistant potato clones were evaluated for their genetic diversity based on 11 isozyme loci and nine microsatellites. A total of 35 allozymes and 42 polymorphic microsatellite fragments was scored for presence or absence. The germplasm was clustered based on the matrix of genetic similarities and the unweighted pair group means analysis of the isozyme and microsatellite data, which were used to construct a dendrogram using NTSYS-pc version 1.7. Twenty-three allozymes and DNA fragments were unique to the wild species. The diploid Solanum species S. berthaultii Hawkes and S. microdontum Bitter formed two distinct phenetic groups. Within S. microdontum, three subgroups were observed. The tetraploid germplasm formed another group, with S. sucrense Hawkes in one subgroup and the cultivated potato and Russian hybrids in another subgroup. Based upon the genetic diversity and the level of late blight resistance, S. microdontum and S. sucrense offer the best choice for strong late blight resistance from genetically diverse sources. This potato germplasm with reported late blight resistance should be introgressed into the potato gene pool to broaden the genetic base to achieve stronger and more durable resistance.

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Timothy A. Rinehart, Brian E. Scheffler, and Sandra M. Reed

Using 14 codominant microsatellite markers that amplify loci across 14 different Hydrangea L. species, we analyzed gene diversity and genetic similarity within Hydrangea. Samples also included Dichroa Lour., Platycrater Sieb. and Zucc., and Schizophragma Sieb. and Zucc. genera to establish their relatedness to Hydrangea species since previous work suggests they may be closely related. Our results support the close affiliation between Macrophyllae E.M. McClint. and Petalanthe (Maxim.) Rehder subsections and their separation from the other Hydrangea species. Most of the Hydrangea species analyzed cluster within their designated sections and subsections; however, genetic distance between species within each subsection varied considerably. Our data suggest that morphological analyses which labeled H. serrata (Thunb.) Ser. as a subspecies of H. macrophylla (Thunb. Ex J.A. Murr.) Ser. are probably more accurate than recent genome size data suggesting H. macrophylla ssp. macrophylla (Thunb.) Ser. and H. macrophylla ssp. serrata (Thunb.) Makino are separate species. Gene diversity estimates indicate that 64.7% of the total diversity is due to differences between species and 49.7% of the overall variation is due to differences between subsections. Low diversity suggests a lack of gene flow between species and subsections and underscores the difficulty in making wide hybrids. Since only 35.3% of the genetic variation is common to all species, unique alleles were used to develop a molecular key for unambiguous species identification and interspecific hybrid verification. Genetic similarity estimates for all 85 samples suggests a roadmap for introgressing horticulturally important traits from different Hydrangea species.

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Xinwang Wang, Robert N. Trigiano, Mark T. Windham, Renae DeVries, Timothy A. Rinehart, James M. Spiers, and Brain Scheffler

The genus Cornus consists of many species, of which C. florida, C. kousa, C. mas, and C. stolonifera are four main ornamental species in North America, Asia, and Europe. For example, over 200 cultivars of C. florida alone have been developed for the nursery industry. Microsatellite loci, or SSR, are useful markers for studying genetic diversity and for creating linkage maps of the various species. The objective of this study was to investigate the genetic diversity between these four Cornus species and eight hybrids. Evaulation of the diversity will be useful in assessing the selection pressure of breeders and/or genetic drift of these dogwood cultivars/lines. Fifteen SSR primer pairs were selected to examine 56 Cornus cultivars and/or lines of the four species and hybrids. The study included 28 C. florida cultivars and lines, 15 C. kousa cultivars and lines, four C. stolonifera cultivars, one cultivar of C. mass and eight hybrids between various Cornus species. An exceptionally high level of diversity was detected among the 56 entries in both the number and size range of SSR alleles. A total of 95 alleles with an average of 7.8 alleles per loci were detected among these 56 genotypes. These selected Cornus cultivars and/or lines could be clustered into four to six subgroups. Some Cornus species were integrated into other species groups, suggesting gene flow between species via the breeding or evolution. SSR markers can contribute to the exploitation of genetic diversity for existing Cornus germplasm. For further study, examination of more SSR loci could explain more completely the diversity among these Cornus cultivars and lines.

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Claudia Cunha, Tana Hintz, and Phillip Griffiths

DNA extractions from 77 snap bean and 2 dry bean cultivars were evaluated for molecular polymorphisms. In total, 100 10-mer oligonuceotide primers were evaluated, and 31 primers that amplified clear and repeatable polymorphisms among bean cultivars were selected. These primers amplified a total of 49 polymorphisms between the cultivars and were used to differentiate the cultivars and evaluate the genetic diversity between them. All cultivars were clustered according to genetic similarities using GenStat 5.0 software, and groupings of pod types were observed when cultivars were separated based on a dissimilarity index. The RAPD polymorphisms will be useful for cultivar determination, seed purity testing and estimation of genetic distances.

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Fenny Dane

American species in the genus Castanea are susceptible to chestnut blight, caused by the Asian fungus Cryphonectria parasitica. This disease spread throughout the natural range of the American chestnut and reduced the species from a timber and nut producing tree to an understory shrub. The lesser known member of the genus, the chinkapin, has also been affected by this disease and a conservation plan is needed. Genetic diversity within and between geographic populations of the Allegheny chinkapin was evaluated to provide baseline genetic information pertinent to conservation of the species. Nuts of Allegheny chinkapin trees from populations in Mississippi, Florida, Alabama, Virginia, and Ohio were collected and evaluated for isozyme and RAPD marker polymorphism. The genetic diversity of these populations will be compared with that of Ozark chinkapin and American chestnut populations. Conservation strategies will be discussed.

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Grey Horton and Jim Luby

This study of genetic diversity in a wild ancestor of the cultivated strawberry was undertaken to describe patterns of variation in nature, assess worth of existing germplasm collections, and identify promising locations for future collection. Previous work reported a similar study of octaploid strawberry ranging east to west across North America. This complementary study focused on variation from north to south in the Rocky Mountains. The morphological diversity of 16 populations of Fragaria virgininia were characterized for morphological and molecular traits. Two clones of each of 133 genotypes from these populations were grown in a common environment in a greenhouse. Eighteen morphological traits, such as leaf area, runner color, and days to flowering, were measured and analyzed with principal components and canonical discriminant analyses. Molecular diversity data were obtained using seven randomly amplified polymorphic DNA primers. Resulting population marker frequencies were also subjected the previously describe anlayses. Differences due to latitude, longitude, and altitude were observed. Implications of the results will be discussed.

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Amnon Levi, Claude E. Thomas, M. Newman, O.U. K. Reddy, X. Zhang, and Y. Xu

Wide phenotypic diversity exists among American heirloom cultivars of watermelon (Citrullus lanatus var. lanatus). However, in published studies, low or no polymorphism was revealed among those heirlooms using isozyme or randomly amplified polymorphic DNA (RAPD) markers. In this study, experiments with inter-simple sequence repeat (ISSR) [also known as simple sequence repeat-(SSR-) anchored primers] and amplified fragment-length polymorphism (AFLP) markers produced high polymorphisms among watermelon heirloom cultivars. ISSR (111) and AFLP (118) markers (229 total) identified 80.2% to 97.8% genetic similarity among heirloom cultivars. The phylogenetic relations based on ISSR and AFLP markers are highly consistent with the parental records available for some of the heirloom cultivars, providing confidence in the dendogram constructed for heirlooms based on similarity values. As compared with RAPD markers, ISSRs and AFLPs are highly effective in differentiating among watermelon cultivars or elite lines with limited genetic diversity.

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Margaret R. Pooler

Many popular crapemyrtle (Lagerstroemia L.) cultivars grown in the United States are interspecific hybrids between L. indica L. and L. fauriei Koehne. The 22 hybrid cultivars released from the U.S. National Arboretum contain primarily genetic material from L. fauriei PI 237884. Examining the genetic diversity ofL. fauriei specimens in the U.S. is valuable because of the historical and economic significance of the species, the increasing interest it is receiving as a source of new cultivars, and its threatened status in the wild. Our objectives were to examine molecular genetic diversity among L. fauriei accessions using Randomly Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphisms (AFLP) markers. Our results indicate: 1) RAPD and AFLP markers are generally consistent in the genetic relationships that they suggest; 2) the L. fauriei germplasm we examined falls into at least three distinct clusters; and 3) the genetic base of cultivated Lagerstroemia could be broadened significantly by incorporating some of this more diverse L. fauriei germplasm into breeding programs.