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Abstract: Comparison of cultures from spores borne on the same basidium has been used for many years in genetic studies of various mushrooms and other Basidiomycetes. No one, however, has isolated both spores from a single basidium of Agaricus bisporus (Lange) Sing., probably because of the comparatively small size of the basiodiospores of this species and of their erratic and unpredictable germination. Observations in our laboratory about a year ago (3) indicate that "pairs" of spores from individual basidia are frequently discharged simultaneously during spore dispersal. These studies also showed that paired spores remain together while falling for at least a meter and can be collected and germinated as pairs on agar media. During the past winter we followed up this lead and attempted to develop a simple procedure for isolating, germinating, and culturing sporelings from pairs of spores collected in this way. This paper describes the techniques that seem preferable. Three problems were encountered: 1. A procedure was required to obtain spore prints with many paired spores free from contamination and sparse enough so that we could be sure that both spores in each pair come from the same basidium. 2. Both spores in many pairs must be made to germinate at approximately the same time. 3. The germinated sporelings must be distributed in a pattern so that they can be readily isolated in pairs or singly.
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