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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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I. Klein, T.M. DeJong, S.A. Weinbaum and T.T. Muraoka

Exposure to photosynthetically active radiation and the consequent effect on leaf mass per unit leaf area (SLW) and nitrogen (percent dry weight and μg·mm-2) allocation within tree canopies was investigated in walnut (Juglans regia `Serr' and `Hartley') trees. Percent contribution of discrete light flux densities below light saturation (100-700 μmol·s-1·m-2) to the total light exposure of individual spurs, exposed up to 9 hour·day-1 to saturating light (>700 μmol·s-1·m-2), was minimal (<1 hour), indicating that individual spurs were either exposed or shaded most of the day. SLW and N content per unit leaf area of individual spurs were highly correlated (second-order polynomial curve fit) with light exposure within the tree canopy, indicating uneven allocation of available N for optimal utilization. Nitrogen expressed as percent dry weight was not correlated with light exposure and SLW. Leaf N content per leaf area was highly correlated (linear fit) with SLW.

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William W. Coates

The ability to remove the pistillate flowers and young nuts from precocious lateral-bearing English walnut (Juglans regia L.) cultivars during the first several years following planting would be useful both in reducing competition with vegetative growth and eliminating potential infection sites for blackline disease (cherry leafroll virus). Applications of ethephon shortly after full bloom were shown to effectively remove all or most pistillate flowers depending upon spray timing and ethephon concentration. Moderate phytotoxicity and reduced seasonal growth limit the usefulness of this technique in the field. Removal of staminate flowers (catkins) prior to pollen release may reduce the excessive pistillate flower abscission of the `Serr' cultivar. Applications of ethephon shortly before the onset of pollen shedding were shown to be ineffective in catkin removal.

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Mohammed A.M. Aly, Robert G. Fjellstrom, Gale H. McGranahan and Dan E. Parfitt

Somatic embryos derived from walnut (Juglans regia L.) ovule tissues were evaluated to determine whether they were of zygotic or maternal origin. Molecular markers were used to permit evaluation at an early stage, before whole plant development. Somatic embryos developed from potentially apomictic `Sunland' and `Cisco' ovule tissue isolated from bagged putatively unpollinated flowers. Phosphoglucomutase (PGM) isozyme analysis showed that all of these embryos, except one from each cultivar, carry the same zymotype as the maternal tissue. However, restriction fragment length polymorphism (RPLP) analysis combined with isozyme evaluation demonstrated that the tested embryos originated from zygotic rather than maternal tissues. This study demonstrates the application of molecular marker analyses, particularly RFLPs, evaluation of somatic embryo origin.

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X. Deng, S.A. Weinbaum, T.M. DeJong and T.T. Muraoka

Abortion of distillate flowers (PFA) in a protandrous cultivar of walnut (Juglans regia L. cv. Serr) was increased by N deficiency. Starch and N concentrations in wood of 2-year-old twigs decreased to minimal levels during abortion of distillate flowers. Nitrogen reserves in woody tissues were reduced by foliar N deficiency, as were concentrations of sugars and N in vacuum-extracted xylem sap. Abortive distillate flowers ceased growth before spur leaves reached 50% of full expansion. PFA may result from transient deficiencies of C and N during the spring flush of growth. Depletion of storage C and N was accentuated before maturation of distillate flowers in this cultivar by the metabolic demands of many catkins, spur growth, and leaf expansion.

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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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L. Carl Greve, Gale McGranahan, Janine Hasey, Ronald Snyder, Kathy Kelly, David Goldhamer and John M. Labavitch

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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John P. Edstrom*, William Krueger and Wilbur Reil

Orchard hedgerow production systems have been used successfully in fruit and nut crops in California for decades to enhance yield, particularly in the early years of production. English walnuts (Juglans regia) are compatible with hedgerow techniques under prime soil conditions but are thought to require deep well drained soil to be commercially productive. Combining the production techniques of micro-irrigation, close spacing, minimal pruning and frequent fertilization in almonds has improved yield substantially on soils exhibiting a shallow, course textured topsoil underlain with a dense clay layer. Paradox hybrid rootstock (J. regia × J. hindsii) has shown greater tolerance to root lesion nematodes and heavier textured or poorly drained soils than Northern California Black (J. hindsii). Fourteen years of evaluation (1986-99) using `Chandler' and `Howard' Ctvs English walnuts in a replicated field trial on marginal soil has shown that 1) yields of 6700 kg·ha-1 (inshell) are attainable under these substandard soil conditions 2) Paradox hybrid rootstock out-yields Northern California Black by 30% on both cultivars tested, 3) kernels of high commercial quality for can be produced for both cultivars and 4) slip plow soil modifications may not improve tree growth, yield or crop quality in drip irrigated walnut hedgerow plantings.

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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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Salih Kafkas, Hakan Ozkan and Mehmet Sutyemez

Turkey has more than 4 million walnut trees (Juglans regia L.), most of which are derived from seedlings, and are nongrafted trees. This characteristic leads to a huge opportunity to select superior walnut genotypes from natural populations for cultivation and for breeding programs. Several selection studies have been performed in the last decades and few genotypes were selected. The goal of this study was to characterize and determine genetic relationships among 21 walnut genotypes with potential in walnut production using amplified fragment length polymorphism (AFLP) and selective amplification of microsatellite polymorphic loci (SAMPL) techniques. Eight primer combinations (six for AFLP and two for SAMPL) were applied to 21 walnut genotypes and a total of 230 bands of which 50.4% of them were polymorphic were obtained. The SAMPL technique was more effective than AFLP in the separation of very closely related genotypes. Genotypes of the pairs `Maras-18' with `Maras-46', `KSU-5' with `Sutyemez-1', `Maras-12' with `Sutyemez-2,' `Kaman-3' with `Kaman-4', and `KSU-11' with `Maras-10' were the most closely related.