Occurrence of genetic segregation in a putative haploid strain of Endomyces fibuliger met by spontaneous sectoring of protoplast fusants

Abstract

Detailed genetic analysis of Endomyces fibuliger, an amylolytic yeast which is homothallic and exists predominantly in the diploid state, has not been performed. From a naturally occurring strain, E. fibuliger 8014 met, a morphological mutant, 193 met, was obtained by u.v. mutagenesis. To obtain a haploid strain suitable for genetic analysis, an intergeneric hybrid between E. fibuliger 193 met and a strain of a closely related dimorphic heterothallic lipolytic yeast, Yarrowia lipolytica, A his1, was produced by mass mating. The intergeneric hybrid was highly unstable in vegetative culture on yeast extract/phosphate/soluble starch/agar media and produced numerous mitotic sectors. Most of the sectors were mitotically unstable. However, one mitotically stable sector, N14i60 met, was obtained which also differed from the strain 193 as gauged by the appearance of DNA bands on pulsed-field gel electrophoresis. The putative haploid strain, N14i60 met, had six bands whilst the mutant 193 met had seven. Ultra-violet treatment of cells of N14i60 met produced 19 auxotrophic mutants. Protoplast fusion between pairs of different mutants showed complementation and the fusants were unstable mitotically and gave unstable aneuploid and stable haploid sectors of parental and non-parental combinations of markers. It is postulated that complementary diploid fusants, which were obtained by protoplast fusion, produced sectors by mitotic non-disjunction. Such a mechanism provides a means to establish a genetic analysis system for E. fibuliger via the parasexual cycle.