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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.2019.tde-20191220-123209
Document
Author
Full name
Janete Magali de Araújo
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1989
Supervisor
Title in Portuguese
Plasmídios conjugativos e fusão de protoplastos em Streptomyces felleus
Keywords in Portuguese
ACTINOMICINA
BACTÉRIAS
FUSÃO DE PROTOPLASTOS
PLASMÍDEOS
PRODUÇÃO
Abstract in Portuguese
Diferentes espécies de Streptomyces, isoladas de amostras de solo do Estado de Pernambuco, mostraram-se produtoras de actinomicina, apresentando formação de pigmento solúvel amarelo-laranja, uma característica de linhagens produtoras desse antibiótico. Três dessas espécies foram ensaiadas em diferentes concentrações de acriflavina e revelaram elevada instabilidade na produção de actinomicina. Em Streptomyces felleus IA-M3079 os mutantes antibiótico-negativos obtidos através do tratamento com o agente intercalante mostraram incialmente um único tipo morfológico, enquanto que os mutantes antibiótico-negativos de S. regensis IA-M3053 apresentaram diferentes características fenotípicas. Foram, também, observadas variantes de S. felleus IA-M3079 com produção de pigmento amarelo-laranja mais intenso que o das linhagens selvagens. As variantes de S. regensis IA-M3053 mostraram-se estáveis para esta característica e as S. felleus IA-M3079 eram instáveis, vindo a desaparecer o pigmento após algumas transferências. Contudo, entre as variantes de S. felleus IA-M3079 destacou-se uma estável com intensa pigmentação. Foi observada, através do fenótipo “pock”, nas três linhagens em estudo - S. felleus IA-M3079, S. parvulus IA-M3042 e S. regensis IA-M3053 – a presença de plasmídios conjugados. A fusão de protoplastos entre S. felleus IA-M3079 e a variante AF-3079 (90/57) mostrou setores recombinantes com formação de pigmento azul-violáceo, que após estabilização e testes antimicrobianos apresentaram elevada atividade antifúngica. As culturas estabilizadas provenientes dos referidos setores segregantes, mostraram estreita similaridade com as parentais quanto às características bioquímicas, micromorfológicas e culturais e ao comportamento eletroforético. O antifúngico produzido pelos setores foi preliminarmente caracterizado através do espectro de absorção na luz ultravioleta.
Title in English
Conjugative plasmids and protoplast fusion in Streptomyces felleus
Abstract in English
Different Streptomyces species, isolated from soil samples of Pernambuco State, Brazil, produce actinomycin and present the formation of yellow-orange solube pigment: a characteristic of strains producing that antibiotic. Three Streptomyces species were tested in different concentrations of acriflavine and showed high instability in the actinomycin production. In Streptomyces felleus IA-M3079, the negative antibiotic mutants obtained through the treatment with the intercalant agent showed a single morphology type, the negative antibiotic mutants of S. regensis IA-M3053 presented different phenotypical features. Besides, it were observed variants of S. felleus IA-M3079 with yellowish-orange pigment production more intensive than that of the wild strains. The S. regensis IA-M3053 variants showed themselves stable for this character and the S. felleus IA-M3079 variants were unstable, disappearing its pigment after some subcultivation. However,among the S. felleus IA-M3079 variants one appeared with intensive and stable pigment. Through the pock phenotype it was observed the presence of conjugative plasmids in the three strains under study: S. felleus IA-M3079, S. parvulus IA-M3042 and S. regensis IA-M3053. The protoplast fusion between S. felleus IA-M3079 and the variant AF-3079 (90/57) produced recombinant sectors with formation of bluish violaceous pigment. Such sectors, after stabilization and antimicrobial tests present high antifungal activity. The purified and stabilized cultures obtained from the recombinant sector showed to be very similar to the parental cultures regarding to biochemical, micromorphological and cultural characteristics as well as to the electrophoretical patterns. The antifugal compound produced by the sector recombinant cultures was characterized through ultraviolet spectra (UV) and by antimicrobial spectra.
 
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Publishing Date
2019-12-20
 
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