Abstract
The protoplast fusion technique was used to obtain useful intraspecific hybrids of Aspergillus usamiimut. shirousamii for making spirits for drinking. Protoplasts were obtained from mycelia of this fungus by using Trichoderma lytic enzyme preparations. Intraspecific hybridization between two auxotrophic mutants of this fungus was achieved by protoplast fusion in the presence of 35% polyethylene glycol as a fusogen. A stable diploid strain was obtained from a heterokaryon strain by treating with 0.1% d-camphor. The diploid strain was treated by benomyl as a haploidizing agent, and many segregants were obtained. Prototrophic segregants were selected and these haploids were confirmed by their conidial size and DNA contents. The diploid strain and some haploids showed high stability and numerous sporulation, producing the same levels of amylase and citric acid as those ofthe original strains. From these results, a technique of intraspecific protoplast fusion was considered to be a useful clue for studying the maintenance of characteristics and homostasis stability and the genetics of parasexual recombination in fungi.
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