The detection of post-meiotic segregation without tetrad analysis in Ustilago maydis

Abstract

Post-meiotic segregation (PMS) results in the formation of mixed genotypes from single meiotic products. A method is described in which single members of tetrads are selected, and these are then tested for their genetic homogeneity. The method is applied to Ustilago maydis using crosses which are heteroallelic for nar 1, the structural gene for nitrate reductase. In the absence of PMS, meiotic products containing a nar+ recombinant are genetically pure (the equivalent of a 6 mutant: 2 wild-type octad). With PMS, a nar+ recombinant clone arises in association with a nar- clone and these are otherwise genetically identical (the equivalent of a 7 mutant: 1 wild type octad). The procedure will make it possible to search for mutant strains which are defective in the correction of mismatched bases in hybrid DNA formed during recombination. Among 26 nar+ recombinants from a control cross, PMS was detected on 3 occasions. In an equivalent cross, both parents were uvs 3, a mutant defective in the excision of pyrimidine dimers from DNA. Among 43 nar+ recombinants, 7 arose from PMS. Thus the frequency of PMS for the nar alleles is about 15% and the excision of pyrimidine dimers is probably unrelated to the repair of mismatched bases in hybrid DNA.