The utility of microsatellite markers to characterize the genetic diversity of a polyploid species with disomic inheritance is often hampered by the impossibility of determining allele frequencies and the complexity of inheritance patterns. The objective of this study was to solve these problems in the allotetraploid Prunus serotina Ehrh. by finding genome-specific primers (i.e., primers that are specific to one of the two genomes that initially formed the species). Sixty-seven microsatellite primers described in cultivated Prunus L. species were tested for cross-amplification in P. serotina, and evidence that conserved markers were genome-specific was found by demonstrating their typical Mendelian diploid inheritance in embryos resulting from controlled crosses. Among the 67 microsatellite markers tested, 26 produced successful amplification and five were genome-specific. No linkage disequilibrium was detected for these loci, but evidence was found for the presence of a null allele at one locus. We found both a high number of alleles per locus (three to 12) and a high mean expected heterozygosity (0.71), which were nonsignificantly different from the number of alleles and estimates of expected heterozygosity calculated for three non-genome-specific markers in the same population. The potential use of these genome-specific markers in population genetic studies is discussed.
Marie Pairon, Anne-Laure Jacquemart, and Daniel Potter
W. Jack Rowe II, Daniel A. Potter, and Robert E. McNiel
Twenty-six purple- or green-leaved cultivars representing 12 species of woody landscape plants were evaluated in the field for defoliation by Japanese beetles (Popillia japonica Newman) over three growing seasons. We further evaluated the hypothesis that, within closely-related plants, purple cultivars generally are preferred over green ones by comparing beetles' consumption of foliage in laboratory choice tests and their orientation to painted silk tree models baited with Japanese beetle lures. Cultivars of Prunus cerasifera Ehrh. and hybrids of that species [e.g., Prunus ×cistena (Hansen) Koehne, Prunus ×blireiana André] were more heavily damaged than nearly all other plants tested. Among maples, Acer palmatum Thunb. `Bloodgood' and A. platanoides L. `Deborah' and `Fairview' were especially susceptible. None of the cultivars of Berberis thunbergii DC, Cercis canadensis L., Cotinus coggygria Scop., or Fagus sylvatica L. were heavily damaged, regardless of foliage color. In the choice tests, purple Norway maples were preferred over green ones in three of four comparisons, but preference varied within the other plant genera. In fact, more beetles oriented to green-leaved tree models than to purple ones. Our results indicate that within a genus, purple-leaved plants do not necessarily sustain more damage than green-leaved ones. Widespread use of certain purple-leaved cultivars of generally susceptible plant species probably contributes to the perception that purpleleaved plants, overall, are preferred. Purple-leaved cultivars of redbud, European beech, smoketree, and barberry, or the purple-leaved Prunus virginiana L. `Canada Red' or Malus ×hybrida Lemoine `Jomarie' may be suitable substitutes for more susceptible purple-leaved plants in landscapes where Japanese beetles are a concern.
Daniel Potter, Fangyou Gao, Giovanna Aiello, Charles Leslie, and Gale McGranahan
The utility of intersimple sequence repeat (ISSR) markers for identification of English or Persian walnut (Juglans regia L.) cultivars was explored. Four cultivars were screened with 47 ISSR primers; eight of these primers, which generated reproducible and informative data, were selected for further study. Two individuals from each of 48 cultivars, including many currently important in the California walnut industry as well as accessions from Europe and Asia, were then examined with the eight ISSR primers. Polymerase chain reaction (PCR) products were separated on agarose gels and stained with ethidium bromide. Fifty-four bands were scored as present or absent in each cultivar; of these, 31 (57%) were polymorphic among the 48 cultivars. Combined data from the eight ISSR primers provided a unique fingerprint for each of the cultivars tested. Fifteen of the cultivars could be distinguished from all others with just one primer, 31 with a minimum of two primers, and two required three primers. Pairwise genetic distances between the cultivars were calculated and a dendrogram was generated using the neighbor-joining algorithm. Some of the groupings in the dendrogram corresponded to groups which, based on known pedigrees, are genealogically closely related. Others included accessions from diverse genetic and/or geographic origins. These results can be attributed to a combination of the limitations of the ISSR method for inferring genetic relationships, on the one hand, and the complex history of walnut cultivar development involving extensive exchange and breeding of germplasm from different geographic regions, on the other.
Gerald S. Dangl, Keith Woeste, Mallikarjuna K. Aradhya, Anne Koehmstedt, Chuck Simon, Daniel Potter, Charles A. Leslie, and Gale McGranahan
One hundred and forty-seven primer pairs originally designed to amplify microsatellites, also known as simple sequence repeats (SSR), in black walnut (Juglans nigra L.) were screened for utility in persian walnut (J. regia L.). Based on scorability and number of informative polymorphisms, the best 14 loci were selected to analyze a diverse group of 47 persian walnut accessions and one J. hindsii (Jepson) Jepson ex R.E. Sm × J. regia hybrid (Paradox) rootstock. Among the 48 accessions, there were 44 unique multi-locus profiles; the accessions with identical profiles appeared to be synonyms. The pairwise genetic distance based on proportion of shared alleles was calculated for all accessions and a UPGMA (unweighted pair group method with arithmetic mean) dendrogram constructed. The results agree well with what is known about the pedigree and/or origins of the genotypes. The SSR markers distinguished pairs of closely related cultivars and should be able to uniquely characterize all walnut cultivars with the exception of budsports. They provide a more powerful and reliable system for the molecular characterization of walnut germplasm than those previously tested. These markers have numerous applications for the walnut industry, including cultivar identification, verification of pedigrees for cultivar and rootstock breeding programs, paternity analysis, and understanding the genetic diversity of germplasm collections.