Techniques and Marker Genes for Use in Macrocyst Genetics with POLYSPHONDYLIUM PALLIDUM

Previous work has shown that genetic exchange occurs in the macrocyst of Polysphondylium pallidum, as in species of Dictyostelium. These studies are extended here. Mutants resistant to six different poisons have been isolated for use as genetic markers. A replica-plating technique has been engineered whereby 14 progeny clones growing on a master plate may be simultaneously transferred to test plates containing individual poisons. Germination percent of macrocysts has been greatly increased by the presence of a growing fungus during the resting stage. These means have been used to analyze crosses showing that: (1) Vegetative amebae are haploid, at least at the three marker loci tested. (2) Amebae emerging from a single macrocyst are identical about 90% of the time. (3) Any single combination of parental markers may emerge from a given macrocyst, and all combinations appear in approximately equal frequencies. These findings suggest that normally one of four nuclei produced by meiosis survives in every macrocyst and that all markers examined are unlinked. (4) About 10% of the macrocysts germinate to give two or more classes of progeny. These may result from the presence of a second zygote in the macrocyst or from the survival of two nuclei after meiosis. (5) If genetic exchange occurs during spore formation or microcyst formation, its frequency is low (<0.01%).