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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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James R. McKenna and Ellen G. Sutter

The use of auxin-impregnated toothpicks stimulated adventitious root formation in genotypes of Juglans `Paradox' that had been backcrossed to J. regia. These genotypes were selected as potential rootstocks because of improved tolerance to cherry leaf roll virus and Phytophthora spp. Other auxin applications including quick dips and talc formulations had little or no effect. The use of toothpicks lowered the concentration of IBA necessary for root initiation compared to previously reported results using quick dips. Toothpicks were inserted transversely into holes drilled 1 to 2 cm above the base of cuttings. Callus and roots always formed at the location of the toothpicks rather than at the base of the cutting. Roots were formed using this method in simple layering, hardwood, and semi-hardwood cuttings. Of all the cuttings that rooted, 90% rooted with toothpicks whereas only 10% rooted using a quick dip. This method may have potential for increasing the efficiency of rooting other difficult-to-root plants.

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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.