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.