METHOD FOR ISOLATION OF MATING-LESS MUTANTS FROM RHODOSPORIDIUM TORULOIDES M 1057, A STRAIN OF MATING TYPE a

Abe, Keiko; Tsuchiya, Eiko; Fukui, Sakuzô; Kodama, Noriko; Kusanagi, Akio

doi:10.2323/jgam.24.287

Cited by 8 publications

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“…Since cellular and nuclear behaviour during the mating process are distinctive, this species is very useful for studying gene expression in relation to cytological changes during sexual differentiation. Isolation of matingless mutants has been reported (Abe et al, 1978), and biochemical analysis in the early stage of sexual differentiation has been attempted (Miyakawa et al, 1982a, b).In the present experiments, self-sporulating progeny were obtained from a sexual cross of haploid A-and a-type strains of R. toruloides. Self-sporulating diploid (SS-type) and aneuploid Aor a-type strains were isolated and classified by genetic and cytological characteristics.…”

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Identification of a Diploid Self-sporulating Cycle in the Basidiomycetous Yeast Rhodosporidium toruloides

Abe

Sasakuma

1986

Microbiology

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~ Self-sporulating diploid and aneuploid cells were identified in the life cycle of Rhodosporidium toruloides by microscopic observation of nuclear behaviour, microphotometry of DNA content, and genetic characterization of progeny. From a sexual cross between haploid A -and a-type strains, diploid progeny with both A and a mating-type loci and aneuploid progeny with either A or a mating-type locus were isolated in addition to haploid progeny. The diploid isolates propagated by budding in yeast form and eventually developed monokaryotic hyphae. The diploid hyphae formed uninucleate teliospores and blastospores from which diploid yeast cells were isolated (self-sporulatirig). Aneuploids carrying the A or a mating-type isolated from a sexual cross and from self-sporulating diploids entered the sexual cycle like haploid cells. The life cycle consisting of sexual and self-sporulating cycles is presented.Sexual activity was recognized in certain isolates of the yeast Rhodotorula glutinis and these were designated as Rhodosporidium toruloides (Banno, 1967). Our studies of the mating process in this organism (Abe et al., 1975(Abe et al., , 1977 can be summarized as follows. Haploid cells designated as A-and a-type propagate by budding. The cells of A-type constitutively secrete a sexual hormone, rhodotorucine A, which arrests the a-type cells at the G, stage and induces the mating activity. The a-type cells then secrete a sexual hormone, rhodotorucine a. As a result, matingtube formation is induced in both a-and A-type cells in response to mating hormones secreted from cells of opposite mating type, and this is followed by cell-to-cell fusion at the tips of the tubes. After fusion, dikaryotic mycelia and teliospores are developed. Since cellular and nuclear behaviour during the mating process are distinctive, this species is very useful for studying gene expression in relation to cytological changes during sexual differentiation. Isolation of matingless mutants has been reported (Abe et al., 1978), and biochemical analysis in the early stage of sexual differentiation has been attempted (Miyakawa et al., 1982a, b).In the present experiments, self-sporulating progeny were obtained from a sexual cross of haploid A-and a-type strains of R. toruloides. Self-sporulating diploid (SS-type) and aneuploid Aor a-type strains were isolated and classified by genetic and cytological characteristics. METHODS Organisms.Haploid strains of Rhodosporidium toruloides IF0 0559-M-919 (mating-type A, Met-, Pan-, pink colony; designated as M-919) and I F 0 0880-M-1057 (mating-type a, Paba-, yellow colony; M-1057) were used as the parental strains. CCY 62-2-22 (mating-type A , Met-, Phe-, white colony) was also used in one cross. The IF0 and CCY strains were kindly provided by Dr I. Banno (Institute for Fermentation, Osaka, Japan) and by D t A.Kochova-Kratochvilova (Institute for Chemistry, Slovak Academy of Science, Bratislava, Czechoslovakia), respectively.Media and cultivation. The yeast extract/sucrose medium (YS medium) and minimal medium (...

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Identification of a Diploid Self-sporulating Cycle in the Basidiomycetous Yeast Rhodosporidium toruloides

Abe

Sasakuma

1986

Microbiology

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“…In this trial, 97 strains of matingless mutants, which were previously isolated from a wild-type strain of R. toruloides M1057, were used. Madia and microorganisms were described in previous papers (1,9).…”

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Sequence of Events in Early Stages of Sexual Differentiation Caused by Rhodotorucine A, a Fungal Sex Hormone: Biological Determi- Nation Using Matingless Mutants Derived From Rhodosporidium Toruloides M 1057

Tsuchiya

Abe

Fukui

1978

J. Gen. Appl. Microbiol.

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Our previous papers described the biological and chemical aspects on the induction of sexual differentiation in life cycle of heterobasidiomycetous yeasts (1-6). For example, two kinds of hormones, rhodotorucine A and a, were essentially required for the transference of asexual cycle to sexual cycle in Rhodosporidium toruloides. Rhodotorucine A is constitutively produced by the cells of R. toruloides M919, mating type A (A cells), into the culture medium, and induces both sexual differentiation and rhodotorucine a formation in the cells of R. toruloides M1057, mating type a (a cells). Recently, rhodotorucine A was isolated from the culture medium of A cells and its chemical structure was determined to be a farnesylpeptide (7). When a cells were incubated with rhodotorucine A, the hormone was immediately adsorbed on the cells, followed by the establishment of intracellular information(s) for sexual differentiation within 15 min at 30° (8). After 5-hr incubation with the hormone, occurrence of following four events was clearly detected in a cells : i) Inhibition of bud formation; ii) inhibition of nuclear duplication; iii) formation of mating tube; and (iv) formation of rhodotorucine a in extracellular space (2). In this paper, we describe one trial to determine the sequence of these four events. In this trial, 97 strains of matingless mutants, which were previously isolated from a wild-type strain of R. toruloides M1057, were used. Madia and microorganisms were described in previous papers (1, 9).Bud formation, mating-tube formation and nuclear duplication were examined as described below. Approximately 104 cells of the mutant (early-log cells) tested were adhered on a cover glass, and incubated with 600 units of rhodotorucine A in 10 ml of culture supernatant of R. toruloides M919 at 27° for 6 hr. Then, according to the method of RoBINow (10) the cells were fixed with AFA reagent and 291

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Topographic and quantitative changes in cell-surface acid-phosphatase during the sexual differentiation of the heterobasidiomycete, Tremella mesenterica

et al. 1984

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METHOD FOR ISOLATION OF MATING-LESS MUTANTS FROM RHODOSPORIDIUM TORULOIDES M 1057, A STRAIN OF MATING TYPE a

Cited by 8 publications

References 4 publications

Identification of a Diploid Self-sporulating Cycle in the Basidiomycetous Yeast Rhodosporidium toruloides

Identification of a Diploid Self-sporulating Cycle in the Basidiomycetous Yeast Rhodosporidium toruloides

Sequence of Events in Early Stages of Sexual Differentiation Caused by Rhodotorucine A, a Fungal Sex Hormone: Biological Determi- Nation Using Matingless Mutants Derived From Rhodosporidium Toruloides M 1057

Topographic and quantitative changes in cell-surface acid-phosphatase during the sexual differentiation of the heterobasidiomycete, Tremella mesenterica

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