Induced mitotic crossing-over inUstilago maydis

Abstract

1. Two different methods of selection were used to obtain stable prototrophic solopathogenic strains ofUstilago maydisfrom haploids with different biochemical requirements. The strains were shown to be heterozygous diploids.2. Two diploids which were examined in detail had four markers in common, but they were in different coupling and repulsion phases. Rare spontaneous segregation was detected in one of the diploids, but a high frequency of segregation was obtained in both after treatment with ultra-violet light. The proportion of segregants amongst the survivors increased with the dose. These auxotrophic segregants were detected by means of the replica plating technique.3. The types of segregant which were obtained from both diploids were consistent with the view that they arose as a result of mitotic crossing-over. After low doses of radiation the reciprocal products of crossing-over were often detected. There was no evidence from the phenotypes of the segregants that haploidization was occurring. The diploidy of a sample of the segregants was confirmed by mating-type tests, and by the fact that they showed further segregation after another dose of radiation.4. A slow-growing unstable segregant recovered after a high dose of radiation proved to be a monosomic strain which consistently reverted to a stable diploid homozygous for one chromosome. It was possible to use this auxotrophic diploid together with a haploid with different biochemical requirements, to synthesize a prototrophic triploid strain. The triploid was much less stable than the diploid strains.5. By means of pathogenicity tests with certain segregants it was possible to distinguish the function of the two loci which control the mating system. Thealocus is responsible for the fusion of haploid sporidia and has no effect on the pathogenicity of the heterokaryon which is under the control of theblocus.6. The effects of ultra-violet light on mitotic crossing-over do not seem to be easily compatible with a copy choice or similar mechanism of recombination.