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Keith Woeste, Gale H. McGranahan and Robert Bernatzky

Twenty-five random decamer primers were used to evaluate the level of polymorphism between Persian walnut and the Northern California black walnut. Sixty-six randomly amplified polymorphic DNA (RAPD) markers were identified using an interspecific walnut backcross population [(Juglans hindsii × J. regia) × J. regia]. Segregation data from these polymorphisms were joined to a previously published set of restriction fragment-length polymorphism (RFLP) marker data to expand the genetic map of walnut to 107 markers in 15 linkage groups.

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Keith Woeste, Gale McGranahan and Robert Bernatzky

A first backcross population of walnuts {[Juglans hindsii (Jeps.) Jeps. × Juglans regia L.] × J. regia} was used to evaluate the correlation between morphological (statistical) and genetic distance during introgression. Five traits based on leaf morphology were identified to quantitate the morphology of the parental species, their F1 hybrids, and the backcrosses to each parent. These traits were used to evaluate the morphological similarity of first backcrosses to J. regia using Mahalanobis' distance. The amount of genomic introgression of each backcross was estimated using 59 randomly amplified polymorphic DNA (RAPD) and 41 restriction fragment-length polymorphism (RFLP) genetic markers that identify polymorphisms between J. regia and J. hindsii. A smaller scaffold set of markers was also identified using published linkage data. The correlation between the measures of morphological and genomic introgression for the first backcrosses was low (0.23) but significant. The results suggest that selection based on morphology during backcrossing will not be an effective way to recover recurrent parent genome.

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Rohollah Karimi, Ahmad Ershadi, Kourosh Vahdati and Keith Woeste

The genetic structure and diversity of natural populations of Juglans regia L. in Iran were characterized using 11 microsatellite loci. A total of 105 individuals from seven populations were sampled. A high level of genetic diversity was observed within populations with the number of alleles per locus (A) ranging from three to 11 (average = 5.73), the proportion of polymorphic loci was 100%, and the expected heterozygosity ranged from 0.598 to 0.848 (average = 0.707). The proportion of genetic differentiation present among populations accounted for 12% of the total variation. Such considerable interpopulation differentiation detected in J. regia L. could have resulted from several factors, including restricted gene flow between populations. Significant departures from Hardy-Weinberg equilibrium were observed for WGA276, WGA32, and WGA321 loci. The deviations were primarily the result of the surplus of heterozygotes. The unweighted pair group method with arithmetic mean cluster analyses based on Nei's unbiased genetic distances separated the seven populations into two main groups.

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Keith Woeste, Douglas Shaw, Gale McGranahan and Robert Bernatzky

We characterized a population of hybrids between English walnut and Northern California black walnut (Juglans regia X J. hindsii) and their backcrosses (BC) using both genomic markers and morphological traits. ANOVA and regression methods were used on three years' data to identify a subset of five variables that describe the morphological variability among backcross populations and their parents (R2 = 0.89). Genomic markers were identified using randomly amplified polymorphic DNA (RAPD). A subset of 60 markers specific to the donor species (J. hindsii) were scored in 50 backcrosses to estimate the percent recipient genome in each evaluated BC. The backcrosses were ranked using each method of evaluation; correlation between the ranks was 0.423 and highly significant. Each evaluation method has advantages but neither was able to reliably identify elite progeny.

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Keith E. Woeste, Gale H. McGranahan and M.N. Schroth

Walnut blight of English walnut (Juglans regia L.), incited by Xanthomonas campestris pv. juglandis (Pierce) Dowson, causes significant crop loss in California. To assess levels of resistance in walnut germplasm, leaves and nuts of mature walnut genotypes were inoculated with X. campestris pv. juglandis. Significant differences were found among cultivars in size and frequency of lesions on leaves and in frequency of abscission of diseased leaves. Cultivars also varied in frequency of abscission of nuts following infection and in marketability of infected nuts. Afthough there was considerable variation in disease levels over 2 years, leaves of PI 159568 consistently received significantly higher disease ratings than leaves of `Chandler' or `Adams'. Nuts of `Adams', `Payne', PI 18256, and `Sinensis 5' abscised less frequently following inoculation than nuts of other cultivars. In addition, the quality of infected nuts that did not abscise was consistently better for PI 18256 and `Sinensis 5'. The rank of cultivars for levels of disease in inoculated leaves was not significantly correlated with the rank of cultivars for frequency of infestation of dormant buds associated with infected foliage. The apparent resistance of walnut germplasm may be affected by the abscission or necrosis of infected tissues.

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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.