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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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Kourosh Vahdati, James R. McKenna, Abhaya M. Dandekar, Charles A. Leslie, Sandie L. Uratsu, Wesley P. Hackett, Paola Negri, and Gale H. McGranahan

Walnuts (Juglans spp.) are difficult-to-root woody plants. The rolABC genes (rolA + rolB + rolC), derived from the bacteria Agrobacterium rhizogenes, have been shown to increase the rooting potential of other difficult-to-root woody plants. We inserted the rolABC genes into somatic embryos of a `Paradox' hybrid (J. hindsii × J. regia) clone PX1 using the A. tumefaciens gene transfer system. A transgenic sub-clone, designated PX1 rolABC 2-2 was selected and compared to the untransformed clone for a variety of phenotypic characteristics, including rooting potential. Transformed and untransformed shoots were budded onto seedling J. regia rootstock in the greenhouse and established in the field. Transformed trees displayed reduced internode length, an increase in lateral branching, and wrinkled leaves. In another test, a commercial persian walnut cultivar J. regia `Chandler' was grafted onto rooted cuttings of both the untransformed and transformed plants. The presence of the rolABC genes in the rootstock had no visible effects on the grafted scion. Several of these trees were excavated from the field and the root systems of each genotype were examined for root number, diameter, and biomass. Trees with the rolABC rootstock had significantly more small diameter roots compared to the controls and less recovered biomass. Tests of the rooting potential of leafy semi-hardwood cuttings for two years resulted in 14% to 59% rooting of the transformed cuttings compared to 51% to 81% rooting of the control. Both transformed hardwood cuttings and microshoots in tissue culture also rooted significantly less (52% and 29% respectively) than untransformed hardwood cuttings and tissue cultured shoots (82% and 54% respectively). Thus, although the rolABC genes induced a shorter internode length and a more fibrous root system (typical of rol-tranformed plants), they were not useful for increasing the rooting potential, and as rootstock they did not affect the phenotype of the scion.

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Gregory T. Browne, Charles A. Leslie, Joseph A. Grant, Ravindra G. Bhat, Leigh S. Schmidt, Wesley P. Hackett, Daniel A. Kluepfel, Reid Robinson, and Gale H. McGranahan

, percentage of root crown length rotted and B , percentage of root rot. On x axis, cal = Juglans californica , nig = J. nigra , reg = J. regia , hin = J. hindsii , maj = J. major , mic = J. microcarpa , and ste = Pterocarya stenoptera . Fig. 2

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Kendra Baumgartner, Phillip Fujiyoshi, Greg T. Browne, Chuck Leslie, and Daniel A. Kluepfel

Armillaria root disease affects orchards in all Juglans regia (Persian walnut)-growing regions of California ( Gardner and Raabe, 1963 ). The causal agent is Armillaria mellea (Basidiomycota, Physalacriaceae), which attacks walnut and other

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John E. Preece and Gale McGranahan

hybrid walnut, and I determined to make the experiment of fertilizing the flowers of the California species with pollen from the Persian” ( Whitson et al., 1914, 1915 , p. 138). The California walnut species has since been identified as Juglans hindsii

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Greg T. Browne, Joseph A. Grant, Leigh S. Schmidt, Charles A. Leslie, and Gale H. McGranahan

(typically Juglans hindsii × J. regia ) ( Mircetich et al., 1998 ). Compared with NCB ( J. hindsii ) and Persian walnut ( J. regia ) rootstocks, PH is more resistant to several species of Phytophthora ( Matheron and Mircetich, 1985b ; Mircetich and

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James R. McKenna and Lynn Epstein

Crown gall, caused by the common soil-borne bacterium Agrobacterium tumefaciens, can be an economic problem in walnut nurseries and production orchards in California. The principal rootstocks used for commercial walnut production in California are the native Northern California black walnut, Juglans hindsii, and “Paradox,” which are interspecific hybrids between a black walnut, primarily J. hindsii, as the maternal parent, and J. regia, the English walnut, as the paternal parent. Recent evidence has shown that some commercial black walnut trees producing Paradox hybrid seedlings are actually hybrids between J. hindsii and two other North American black walnut species, J. major and J. nigra. Here, we document that there was a higher incidence of crown gall on Paradox (J. hindsii ×J. regia) than on J. hindsii in three sites with natural soil inoculum. Paradox seedlings (with a female parent that was primarily J. hindsii with some J. nigra) inoculated with A. tumefaciens on the roots during transplanting had a higher incidence of crown gall than either J. hindsii or J. regia. When stems were inoculated with A. tumefaciens, J. hindsii ×J. regia populations had significantly larger galls than either J. hindsii or J. regia. Similarly, in stem inoculations on four out of six Paradox genotypes with a hybrid black walnut maternal parent, the progeny produced significantly larger galls than either J. hindsii or J. regia. However, two Paradox populations from black walnut hybrids that contained J. major, J. nigra, and J. hindsii produced galls that were no different in size than in the black walnut species and J. regia. Results suggest that J. regia and black walnut species are less susceptible to crown gall than most Paradox populations.

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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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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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Janet Caprile and Steve Grattan

In areas of California with a high incidence of walnut blackline disease, walnut orchards are increasingly being planted on English walnut (Juglans regia) rootstocks that are tolerant to the virus. There is limited documentation on the salt tolerance of this rootstock. This work was done to quantify the response of English walnut rootstocks to high boron (B), chloride (Cl), and sodium (Na) and to compare this to the more common rootstocks, Northern California Black (J.hindsii) and Paradox (J. hindsii × J. regia). The trial was configured as a randomized complete-block design with 20 plots. Plots consisted of three proximate, matched `Chandler' trees, each on a different rootstock. Leaf samples over a 2-year period showed that trees on the English rootstock had a significantly higher salt uptake than trees on Paradox, which had a significantly higher uptake than trees on Black. Bark cores showed little difference in B uptake between any of the rootstocks. Bark cores also showed that all rootstocks accumulated significantly more B than the scion portion of the tree, indicating that B transport may be limited by the graft union. Trees on English rootstock had significantly more B in the scion bark cores than trees on either Black or Paradox, indicating that rootstocks with a J. hindsii parent may be better at reducing salt transport across the graft union.