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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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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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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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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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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

trees Plant Dis. 69 1039 1041 Matheron, M.E. Mircetich, S.M. 1985c Seasonal variation in susceptibility of Juglans hindsii and Paradox rootstocks of English walnut trees to Phytophthora citricola Phytopathology 75 970 972 Matkin, O.A. Chandler, P

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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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Richard P. Buchner*, Allan Fulton, Bruce Lampinen, Ken Shackel, Terry Prichard, Larry Schwankl, Sam Metcalf and Cayle Little

Ninth leaf California Chandler Walnuts (Juglans regia) on Northern California Black (Juglans hindsii) or Paradox (English/black hybrid) rootstock were irrigated to achieve three levels of Midday Stem Water Potential (MSWP). Target potentials were: 1) low water stress (average MSWP of -3.2 bars); 2) mild water stress (average MSWP of -6.2 bars); and 3) moderate water stress (average MSWP of -7.3 bars). Stem Water Potential was measured midday (12-4 pm) by placing leaves inside water impervious, light blocking foil bags. Leaves remained bagged for at least ten minutes to achieve equilibrium. Bagged leaves were removed, placed inside a pressure chamber and stem water potential was measured at endpoint. Data are presented for the 2002 and 2003 seasons. Withholding irrigation water had a significant impact on `Chandler' growth, productivity, and profitability particularly on young, vigorously growing trees. Chandler/Black appears to be more tolerant to water stress compared to Chandler/Paradox For Chandler on Paradox, water stress significantly reduced growth, yield, price per pound, percent edible kernel, and resulted in darker kernels. In addition, water stress significantly increased the total percent offgrade. Withholding irrigation does not appear to be a good strategy in young, vigorously growing `Chandler' orchards. Mature trees and trees grafted onto Northern California black rootstock may be more tolerant of moisture stress.

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J.R. McKenna and E.G. Sutter

Experiments with field-grown hybrid Paradox (Juglans hindsii × J. Regia) walnut trees were conducted to assess the effects of stock plant water status, auxin application method, and the addition of spermine on adventitious root formation in stem cuttings. A 2-fold increase in rooting was noted when semihardwood cuttings were collected from dry (midday Ψw = –1.3 MPa) stock plants compared to the same trees six days later when fully hydrated (midday Ψw = –0.6 MPa). Spermine, when combined with potassium indolebutyric acid (KIBA) and applied as a quick dip, enhanced the rooting percentage in hardwood cuttings (54%) compared to controls treated with KIBA alone (18%). Spermine had no effect when it was applied together with KIBA using a toothpick application, producing 65% rooting compared to controls which had 75% rooting. By itself, spermine had no effect on rooting. The toothpick method for applying rooting compounds resulted in significantly higher rooting percentages for hardwood cuttings, but not for semihardwood cuttings. Combining spermine with KIBA had no effect on rooting of semihardwood cuttings.

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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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Xiaobo Song, Shengke Xi, Junpei Zhang, Qingguo Ma, Ye Zhou, Dong Pei, Huzhi Xu and Jianwu Zhang

excellent walnut rootstocks for walnut industry growing in China. Origin ‘Zhong Ning Sheng’ (ZNS) is the result of a cross between JH-8202 ( Juglans hindsii ) and LNJR-01 ( Juglans regia ) performed at a seedling field in Luoning County, Henan, China. The