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  • Author or Editor: Gale McGranahan x
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Insecticidal crystal protein fragments (ICPFs) of Bacillus thuringiensis (Bt) encoded by cryIA(c) gene were shown in diet incorporation studies to be lethal to codling moth (CM; Cydia pomonella) the key insect pest for walnut. However transformed walnut tissues expressing cryIA(c) with Bt codon usage patterns and native DNA sequence revealed very low levels of expression in planta. To correct this problem synthetic versions of one of these genes, cryIA(c) was used to transform walnut tissue. A total of 61 individual transgenic embryo lines were obtained. 34% of these lines (21/61) were high expressors (“class A”) demonstrating 80 to 100% mortality of first in star CM larvae and displaying no further larval development. Twelve clones (20%) were designated “class B” and these showed a marked retardation of larval development and a mortality between 40 to 79%. Embryos from the remaining 28 lines designated “class C” (46%). although transformed, were indistinguishable from the control (untransformed embryos) and showed a mortality of 0 to 39%.

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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.

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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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The variation in polyunsaturated fatty acid content of walnut (Juglans regia L.) oils was determined by analysis of samples isolated from specimens growing in four germplasm collections [California (55 cultivars), Washington (64 seedlings), China (12 cultivars), and France (20 cultivars)]. In addition, the impact of within-state geographic differences on oil composition was examined by comparing samples from three California cultivars (`Ashley', `Hartley', and `Franquette') grown in three locations. Local environmental effects on oil composition of `Chico' were also examined by comparing 1) samples collected from shaded and sun-exposed locations of the same trees and 2) samples collected from trees subjected to three irrigation regimes. Polyunsaturated fatty acid content, as a percentage of total fatty acids, ranged from 47.2% in nuts from PI 142323 from France to 81.0% in `Ashley' from California. However, our data indicate that environment, genotype, nut maturity, and their interactions all contribute significantly to variation in the degree of unsaturation of walnut oil.

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Species of Phytophthora are serious soilborne pathogens of persian (english) walnut, causing crown and root rot and associated production losses worldwide. To facilitate the development of improved walnut rootstocks, we examined resistance of 48 diverse clones and seedlings of Juglans species to P. cinnamomi and P. citricola. Plants were micropropagated, acclimatized to a greenhouse environment, and then exposed to the pathogens in artificially infested potting soil mix. Inoculated plants, as well as noninoculated controls, were subjected to soil flooding for 48 hours every 2 weeks to facilitate infection by the pathogens. Two to 3 months after inoculation, resistance to the pathogens was assessed according to the severity of crown and root rot. Clonal hybrids of J. californica × J. regia were highly susceptible to the pathogens (means 52% to 76% root crown length rotted), while several clones of J. microcarpa × J. regia were significantly less susceptible (means 8% to 79% crown length rotted). Among clones of other parentages tested, including: J. microcarpa, (J. californica × J. nigra) × J. regia, J. hindsii × J. regia, (J. hindsii × J. regia) × J. regia, [(J. major × J. hindsii) × J. nigra] × J. regia, and J. nigra × J. regia, responses varied, but tended to be intermediate. When ‘Serr’ scions were budded or grafted on J. microcarpa × J. regia clone ‘RX1’ or Paradox (J. hindsii × J. regia) seedling rootstocks in a commercial orchard infested with P. cinnamomi, all trees on ‘RX1’ remained healthy, whereas only 49% of those on Paradox survived. Thus, useful resistance to Phytophthora is available among J. microcarpa × J. regia hybrids and is evident in ‘RX1’ rootstock.

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