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Warren F. Lamboy, Jing Yu, Phil L. Forsline and Norman F. Weeden

One of the primary progenitors of the cultivated apple is Malus sieversii L., a species native to the forested regions of central Asia. Despite the horticultural importance of M. sieversii, little is known about genetic variation in this species. In this study, allozyme diversity at 18 loci was determined for 259 seedlings belonging to 31 sib families, each consisting of the set of offspring from a different open-pollinated maternal (seed) parent. Maternal parents belonged to 14 populations from four geographic regions. Genetic diversity statistics were computed from the resulting allele and phenotype frequencies. Cluster analysis of sib families showed that there was some grouping based on geographic region, but 16 of the sib families were most closely related to sib families from other regions. Analysis of molecular variance (AMOVA) indicated that 85% of the enzyme variability was due to differences among sib families within populations and 15% was due to differences among regions. No variability could be assigned to differences among populations within regions. In addition, no alleles were found that were fixed in a region and unique to that region. These results suggest that plants belonging to M. sieversii effectively form a panmictic population. Consequently, a thorough sampling of a few large populations will efficiently capture most of the genetic diversity present in wild M. sieversii.

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Hongwen Huang, Fenny Dane and Tom Kubisiak

The population structure and genetic diversity of American chestnut trees collected in nine states along the natural range of the species was evaluated using 20 isozyme loci. Genetic heterozygosity (Ht:Nei, 1978) ranged from 0.089 in the Georgia and 0.094 in the North Carolina population to 0.172 in the northernmost (Connecticut) and 0.181 in the southernmost (Alabama) population. Four populations (Pennsylvania, New York, Virginia, and Alabama) were selected for RAPD analysis using 22 loci randomly distributed across the chestnut genome. The highest level of heterozygosity was in the Alabama population. UPGMA phenograms generated for the isozyme and RAPD markers using Nei's genetic identity showed similar genetic relationships among American chestnut populations.

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Morshidi Maimunah, R.M. Manshardt and Francis Zee

Populations of wild Carica papaya, previously designated as Carica peltata, were sampled from its native range on the Caribbean coast of Central America (Mexico, Belize, Guatemala, Honduras) and cultivated Carica papaya from both Central and South America were examined for isozyme variability. Thirteen loci from nine enzyme systems (Pgm, Pgi, Idh, Mdh, 6Pgd, Ugpp, Skdh, Aco, Tpi) were scored for all populations. Ten loci were polymorphic and a total of 31 alleles were detected. Isozyme genotypes as determined through segregation analysis were used in the genetic interpretation for eight loci and 18 alleles while six additional loci and 13 alleles were postulated on the basis of phenotypic variation found throughout the species. Nei's genetic identity, I, for both cultivated and wild Carica papaya was >0.9, which is consistent with conspecific populations. Wild papaya populations from different geographic areas appear more related to one another than to domesticates in the same geographic region.

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Warren F. Lamboy and Christopher G. Alpha

Curators of plant genetic resources collections must preserve germplasm possessing known useful characteristics as well as material displaying general genetic diversity. In order to ensure that both types of germplasm are included in a collection, germplasm curators require three fundamental types of information about each accession: taxonomic identity, genetic identity, and genetic relationship. Because simple sequence repeat DNA fragments (SSRs) have been successfully used to determine the genetic identity of grape clones, we conducted a study to determine if SSRs would supply all three types of information for the accessions in the cold-hardy Vitis (grape) germplasm collection. SSR fragments were amplified at six different loci for 23 accessions of cold-hardy grape spanning the range of species diversity in the collection. The minimum number of different alleles found at a locus was 9; the maximum was 26. Heterozygosity values ranged between 0.565 and 0.783, while gene diversity values were in the range 0.785 to 0.944. Two hundred fifty-two pairs of plants out of a possible 253 could be distinguished by their SSR profiles. Nei's genetic identities were computed between all pairs of plants and used in a UPGMA cluster analysis. The relationships obtained did not correspond well to expected relationships based on geography and taxonomy. Four species of grapes were represented by two or more accessions in this study. No DNA fragments found at these six loci served to unambiguously distinguish one species from another. Thus, SSR fragments from the six loci studied were useful in determining genetic identity of accessions, but were not helpful in determining genetic relationships or taxonomic identities. We are searching for additional loci that are informative for these types of information. Meanwhile we highly recommend SSRs for determining genetic identity in germplasm resources collections.

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Hongwen Huang, Desmond R. Layne and Thomas L. Kubisiak

Twelve, 10-base primers amplified a total of 20 intense and easily scorable polymorphic bands in an interspecific cross of PPF1-5 pawpaw [Asimina triloba (L.) Dunal.] × RET (Asimina reticulata Shuttlew.). In this cross, all bands scored were present in, and inherited from, the A. triloba parent PPF1-5. Nineteen of the 20 bands were found to segregate as expected (1:1 or 3:1) based on chi-square goodness-of-fit tests, and were subsequently used to evaluate genetic diversity in populations of A. triloba collected from six states (Georgia, Illinois, Indiana, Maryland, New York, and West Virginia) within its natural range. Analysis of genetic diversity of the populations revealed that the mean number of alleles per locus was A = 1.64, percent polymorphic loci was P = 64, and expected heterozygosity was He = 0.25. No significant differences were found among populations for any of the polymorphic indices. Partitioning of the population genetic diversity showed that the average genetic diversity within populations was Hs = 0.26, accounting for 72% of the total genetic diversity. Genetic diversity among populations was Dst = 0.10, accounting for 28% of the total genetic diversity. Nei's genetic identity and distance showed a high mean identity of 0.86 between populations. Genetic relationships among the populations examined by unweighted pair-group mean clustering analysis separated the six populations into two primary clusters: one composed of Georgia, Maryland, and New York, and the other composed of Illinois, Indiana, and West Virginia. The Georgia and Indiana populations were further separated from the other populations within each group. This study provides additional evidence that marginal populations within the natural range of A. triloba should be included in future collection efforts to capture most of the rare and local alleles responsible for this differentiation.