Rhodosporidium BANNO: Inaktivierende Wirksamkeit von ultraviolettem Licht und N-Methyl-N′-nitro-N-nitrosoguanidin in der Hefephase

Böttcher, F.; Ia, Samsonova

doi:10.1002/jobm.3630170405

Cited by 8 publications

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References 14 publications

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“…Die Mutagenisierung erfolgte, wie bei BOTTCHER u. SAMSONOVA (1977) (KIMBALL 1970, LEE u. JONES 1973, BURKE u. FANOMAN 1975, KEE u. HABER 1975. Die MNNG-induzierte Mutagenese findet wie bei Bakterien (CERDL-OLMEDO et al 1968, GUEROLA et al 1971, SICCARDI et al 1976) bevorzugt in den DNA-Replikationsgabeln statt, was zur Entstehung von Mehrfachmutanten, zur Co-Mutation der im Bereich der Replikationsgabeln liegenden Gene fuhrt (DAWES et al 1977 OLTMANNS (1964) und von uns (BOTTCHER et al 1972a, 1972b…”

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RhodosporidiumBANNO: Dosiseffektbeziehungen, Mutageneffektivität und Mutantenspektrum bei der Induktion Auxotrophie verursachender Mutationen durch ultraviolettes Licht und N‐Methyl‐N′‐nitro‐N‐nitrosoguanidin

Böttcher

Samsonova

1978

J of Basic Microbiology

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The kinetics, efficiency, and specificity of induction of forward mutations to auxotrophy by ultraviolet light (UV) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was examined in stationary phase cells of Rhodosporidium (Rhodotorula) wild strain Rgl.In comparison t o the spontaneous level the frequency of auxotrophic mutants was increased more than 1000 times by both mutagens, however, the mutagenic efficiency of MNNG was higher than that of UV. We found that the forward mutation rate is a linear function of the applicated UV and MNNG doses in the range t o 600 J m-2 or 25 mM X min, respectively.The 35 studied biosynthetic pathways to amino acids, purines, pyrimidines, and vitamins are genetically blocked at different frequencies, but there is not any significant difference between UV and MNNG induced frequencies of mutants with a specific requirement. However, in difference to the approximately equal distribution of the MNNG-induced ~i c mutants among the genetic blocks of the tryptophan-nicotinamide pathway, UV-induced nic mutants occurred with a higher frequency in the genes of the tryptophan pyrrolase and the 3-hydroxykynureninase than in the genes of the other enzymes of the pathway. Kurzwelliges ultraviolettes Licht (UV) und N-Methyl-N'-nitro-N-nitrosoguanidin (MNNG) sind effektive und hiiufig genutzte Mutagene, die sich in ihren Interaktionen niit DNA, in den induzierten Mutagenesemechanisnien, im Typ und in der Lokalisation der erzeugten Mutationen unterscheiden ( BRESLER 1976, DRAKE u. BALTZ 1976, YRAKASH u. SHERMAN 1973, SOIFER 1976). Im Rahnien von Experimenten zur Gewinnung von Mutanten fur genetische und biochemische Arbeiten fuhrten wir vergleichende Untersuchungen uber die Dosiseffektbeziehungen, die Effektivitat und Spezifitat beider Mutagene bei der Induktion von Auxotrophie verursachenden Mutationen an einem Hefestamm von Rhodosporidium toruloides durch. Vorlaufige Befunde iiher die UV-Induktion von auxotrophen Mutanten bei diesem Stamin wurden bereits publiziert (BOTTCHER et al. 1972a). Material und MethodenDer in den Versuchen verwendete Stamm Rgl wurde aus einer im nordostlichen Teil der DDR entnommenen olhaltigen Erdprobe isoliert. Nach KOCKOV~-KRATOCHVILOV~ u. WEGENER (1973) handelt es sich um einen Stamm von Rhodotorula glutinis (FREs.) HARRISON. Rgl ist prototroph und kann %-Alkane als C-Quelle verwerten. Er bildet mit dem Stamm IF0.0559 (Pearungstyp ,.A"), nicht aber mit dem Stamm IF0.0880 (Paarungstyp ,,a") von Rhodosp. toruloides BANNO ein schnallentragendes dikaryotisches Hyphenmycel mit dem fur diese Art typischen Teliosporen. Demnach ist Rgl die Hefephase von Rhodosp. toruloides und repriisentiert den Paarungstyp ,,a".

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RhodosporidiumBANNO: Dosiseffektbeziehungen, Mutageneffektivität und Mutantenspektrum bei der Induktion Auxotrophie verursachender Mutationen durch ultraviolettes Licht und N‐Methyl‐N′‐nitro‐N‐nitrosoguanidin

Böttcher

Samsonova

1978

J of Basic Microbiology

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Hybridization of yeasts by protoplast fusion: Early events after fusion in Pichia guilliermondii

Klinner

Böttcher

1984

J of Basic Microbiology

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The efficiency of protoplast fusions in haploid strains of P. guilliermondii was dependent on the strain used. I n the primary fusion colonies, a large proportion of non-dividing cells was sometimes observed. While in two strains stable hybrids occurred immediately after fusion, one strain showed a longer-lasting heterokaryotic state.It is now well established that stable hybrids can be constructed by protoplast fusion i n yeasts (FERENCZY 1981, ALFOLDI 1982). Little is known, however, about the processes involved, which start with the fusion of the cytoplasrriic niembranes of the protoplasts and end with the formation of stable hybrid cells. They include, for instance, the formation of an united cytoplasm froni the primary chiniaeric fusion product, the regeneration of the cell wall, the fusion of the nuclei within the heterokaryon and the formation of a typical yeast cell capable of normal reproduction by budding. Some of these processes may be similar to corresponding sexual processes, but they are certainly not so tightly ordered. Obviously, protoplast fusion differs from sexual hybridization : The preparation of the cells for mating by pheromones, which leads to an arrest in the GI-phase of the cell cycle, is absent. BURTON 1954, YRAHL et al. 1980), utilize n-alkanes as well as carbohydrates. In this paper we show that the formation of hybrid nuclei in the heterokaryons takes place in a non-identical way in different strains. Materials and methodsStrains: Diauxotrophic mutants of several strains of P. guilliermondii were used ( Table 1). The mutations were induced by UV irradiation as described by BOTTCHER and SAMSONOVA (1977, 1978).Single markers reverted with a frequency lower than lo-*.Media: Solid complete medium (SCM) contained 2% glucose, 1% peptone, 2.5% malt extract, 1% yeast extract,, 2 O 4 , agar. Liquid complete medium contained 2% glucose, 2.5% ,peptone and 1% yeast extract. Solid minimal medium (SMM) was composed as described by BOTTCHER and SAMYONOVA (1978), except that 2 mg/l biotin was added to allow rapid growth of P.

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Auswirkungen von Mutanteninduktion und Protoplastenfusion auf den DNA‐ und Proteingehalt von Pichia guilliermondii

Büttner

Klinner

Birnbaum

et al. 1985

J. Basic Microbiol.

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The DNA content per cell of two tested auxotrophic mutants of Pichia guilliermondii was diminished in comparison to their wild-type strains. After protoplast fusion between these mutitnta the majority of hybrids did not show the expected amount of DNA. In some hybrids it was only slightly higher than in the wild-type strains, although heterozygosity for markers of three chromosomes could be established.The protein content per cell of most strains was positively correlated wit.h the DNA content, but generally more affected as the DNA content by the induction of mutations and protoplast fusion. The protein/DNA ratio of the two mutants was strongly reduced, while this ratio was somewhat enhanced in one of the hybrids in comparison to the wild-type strains and the majority of hybrids. Apparently, thc reduced protein content per cell in the mutants seemed to be partially compensated in the hybrids. FXRENCZY 1977, BOTTCHER et al. 197.8, MARAZ et al. 1978, VALLIN u. FERENCZY 1978. Zur Hybridisierung von Mikroorganismen wurde in den letztenDariiber hinaus sind biochemische Konsequenzen der Protoplastenfusion bisher wenig untersucht worden (ANNE u. PEBERDY 1981).Wir haben bei der Futterhefe Pichia guilliermondii die Korrelation zwischen DNAund Proteingehalt pro Zelle bei Fusionshybriden und ihren Ausgangsstammen untersucht. Wciterhin wixrde in diesen Stammen das Muster ausgewahltcr Enzyme bestimmt. I n der vorliegenden Arbeit sind zunachst die Ergehnisse der genetischen Charakterisierung sowie der DNA-und Proteinbestimmungen beschrieben. Material und Methoden

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Rhodosporidium BANNO: Inaktivierende Wirksamkeit von ultraviolettem Licht und N-Methyl-N′-nitro-N-nitrosoguanidin in der Hefephase

Cited by 8 publications

References 14 publications

RhodosporidiumBANNO: Dosiseffektbeziehungen, Mutageneffektivität und Mutantenspektrum bei der Induktion Auxotrophie verursachender Mutationen durch ultraviolettes Licht und N‐Methyl‐N′‐nitro‐N‐nitrosoguanidin

RhodosporidiumBANNO: Dosiseffektbeziehungen, Mutageneffektivität und Mutantenspektrum bei der Induktion Auxotrophie verursachender Mutationen durch ultraviolettes Licht und N‐Methyl‐N′‐nitro‐N‐nitrosoguanidin

Hybridization of yeasts by protoplast fusion: Early events after fusion in Pichia guilliermondii

Auswirkungen von Mutanteninduktion und Protoplastenfusion auf den DNA‐ und Proteingehalt von Pichia guilliermondii

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