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The mode of reproduction and the sex ratio of Aphelenchoides besseyi were studied in vitro using known numbers of larvae as inocula and the fungus Aureobasidium pullulans as a host. Inoculations with single larvae demonstrated that the nematode does not reproduce parthenogenetically. It is proposed that sex ratio of an obligate amphimictic nematode can be estimated by the equation: Pr = 1-mn-(1-m)n, where 'Pr' is the probability of an inoculum to reproduce, 'n' the number of larvae employed in the inoculum and 'm' being the male ratio of the population. Based on the experimentally obtained 'Pr' and 'n', male ratio ('m') of A. besseyi studied was estimated to be 22.7%. Results obtained by analyses of the adult nematodes from infested rice grains at "hard dough" stage were similar to the estimated value. Similar analyses of the nematodes reared on A. pullulans at 30°, however, showed that the male ratio increased with the age of the culture. It is suggested that poor food supply may favor the male differentiation and/or shorten the relative life span of the females.