Methods for the isolation and analysis of plasmodial mutants in<i>Physarum polycephalum</i>

Cooke, D.; Dee, Jennifer

doi:10.1017/s0016672300015202

Cited by 110 publications

(89 citation statements)

References 12 publications

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“…All strains were largely isogenic with the Colonia isolate (Adler & Holt, 19746;Cooke & Dee, 1975). Principal amoebal strains are shown in Table 1.…”

Section: E T H O D Smentioning

confidence: 98%

Regulation of Development by the matA Complex Locus in Physarum polycephalum

et al. 1989

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1347 ~The transformation of uninucleate amoebae of Physarum polycephalum into multinucleate plasmodia is under the control of a multiallelic mating-type locus, matA. Plasmodium formation usually occurs only when amoebal fusion brings together two different matA alleles within a single cell; such plasmodium formation is termed crossing. Mutations (gadA) in the matA locus permit haploid amoebae to self, that is to form plasmodia in clonal cultures without amoebal fusion. By constructing diploid amoebae heterozygous for gadA alleles, it was shown that the mutant alleles gadA.5 and gadAl1 I were each dominant to gadA+. Non-selfing revertants of gadA mutants may be generated as a result of further mutations (npf) within the matA locus. Sixty-one revertants of this type were studied, derived from three matA3 gadA mutants. Some revertants were able to cross with a matA3 gadA+ tester strain. These 'class I' revertants failed to cross with one another or with similar revertants of two matA2 gadA mutants; they appear all to lack a single gene function (npfC+) that is necessary in plasmodium development. Other revertants failed to cross with the matA3 gadA+ tester. These 'class 11' revertants also failed to cross with one another, but showed several different patterns of crossing when mixed with other strains. The behaviour of some of the class I1 revertants suggests that they lack a further gene function (npfB+) necessary in plasmodium development. The gadA mutations may affect a cisacting regulator of npfB+.

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“…All strains were largely isogenic with the Colonia isolate (Adler & Holt, 19746;Cooke & Dee, 1975). Principal amoebal strains are shown in Table 1.…”

Section: E T H O D Smentioning

confidence: 98%

Regulation of Development by the matA Complex Locus in Physarum polycephalum

et al. 1989

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“…The amoebal strains used were LU859 (matA2 matB2 matC2; Cooke, 1974), CH508 (matA2 matB3 matC2 ;Youngman et al, 1979) and LU648 (matA1 matB2 matC2; Cooke & Dee, 1975). Stocks were maintained on lawns of Escherichia coli on liver infusion agar at 26 "C (Bailey et al, 1987).…”

Section: Methodsmentioning

confidence: 99%

Cellular events during sexual development from amoeba to plasmodium in the slime mould Physarum polycephalum

Bailey

Anderson

Dee

1990

Journal of General Microbiology

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Time-lapse cinematography and immunofluorescence microscopy were used to study cellular events during amoebal fusions and sexual plasmodium development in Physarum polycephulum. Amoeba1 fusions occurred frequently in mixtures of strains heteroallelic or homoallelic for the mating-type locus mafA, but plasmodia developed only in the matA-heteroallelic cultures. These observations confirmed that mafA controls development of fusion cells rather than cell fusion. Analysis of cell pedigrees showed that, in both types of culture, amoebae fused at any stage of the cell cycle except mitosis. In mafA-heteroallelic fusion cells, nuclear fusion occurred in interphase about 2 h after cell fusion; interphase nuclear fusion did not occur in mafA-homoallelic fusion cells. The diploid zygote, formed by nuclear fusion in mafA-heteroallelic fusion cells, entered an extended period of cell growth which ended in the formation of a binucleate plasmodium by mitosis without cytokinesis. In contrast, no extension to the cell cycle was observed in mafA-homoallelic fusion cells and mitosis was always accompanied by cytokinesis. In rnatA-homoallelic cultures, many of the binucleate fusion cells split apart without mitosis, regenerating pairs of uninucleate amoebae; in the remaining fusion cells, the nuclei entered mitosis synchronously and spindle fusion sometimes occurred, giving rise to a variety of products. Immunofluorescence microscopy showed that rnatA-heteroallelic fusion cells possessed two amoebal microtubule organizing centres, and that most zygotes possessed only one; amoebal microtubule organization was lost gradually over several cell cycles. In mafAhomoallelic cultures, all the cells retained amoebal microtubule organization.

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“…For the isolation of nutritional mutants in amoebae, growth on agar-based fully defined media will be essential. However, with the liquid defined media already available, it is possible to test for nutritional requirements in amoebae carrying mutations identified in auxotrophic plasmodia (Cooke & Dee, 1975).…”

Section: H R M C C U L L O U G H a N D J D E Ementioning

confidence: 99%

Defined and Semi-defined Media for the Growth of Amoebae of Physarum polycephalum

McCULLOUGH

Dee

1976

Journal of General Microbiology

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Amoebae of the true slime mould Physarum polycephalum were cultured in two fully-defined liquid media containing amino acids, glucose, three vitamins and a buffered salts solution. Absolute requirements were demonstrated for methionine, haematin, .thiamine and biotin, all of which were known to be specific requirements of the plasmodial stage. Methods are described for large-scale culture in three semi-defined media.

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Methods for the isolation and analysis of plasmodial mutants inPhysarum polycephalum

Cited by 110 publications

References 12 publications

Regulation of Development by the matA Complex Locus in Physarum polycephalum

Regulation of Development by the matA Complex Locus in Physarum polycephalum

Cellular events during sexual development from amoeba to plasmodium in the slime mould Physarum polycephalum

Defined and Semi-defined Media for the Growth of Amoebae of Physarum polycephalum

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