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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.2019.tde-20191220-125145
Document
Author
Full name
Iara Fino Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1996
Supervisor
Title in Portuguese
Caracterização genética por RAPD de recombinantes em Trichoderma pseudokoningii
Keywords in Portuguese
CARACTERIZAÇÃO GENÉTICA
DNA RECOMBINANTE
FUNGOS
MARCADOR MOLECULAR
MUTANTES
Abstract in Portuguese
No presente trabalho foram realizados estudos genéticos no fungo Trichoderma pseudokoningii, utilizando-se duas linhagens contrastantes e isoladas independentemente, a EST-3 e A M. Na EST-3 foram obtidos cinco mutantes auxotróficos simples e quatro mutantes com dupla deficiência, empregando-se para tal, luz ultravioleta. A taxa de mutação foi baixa (0,001%), sugerindo ser essa linhagem muito resistente a irradiação ultravioleta, ou possuir um sistema de reparo muito eficiente. Dois mutantes auxotróficos duplos, originados da EST-3, foram utilizados nos experimentos de fusão de protoplastos com dois mutantes auxotróficos de linhagem em M, sendo um deles duplo e o outro com quatro marcadores. Colônias recombinantes foram recuperadas diretamente de produtos de fusão, em uma baixa freqüência (1%), indicando reduzida freqüência de recombinação na parameiose entre as duas linhagens. Os mutantes parentais e seus recombinantes foram analisados pelo RAPD com a utilização de dez primers decâmeros arbotrários. Os padrões dos parentais foram distintos com todos os dez primers, sugerindo uma divergência genética entre as duas linhagens utilizadas. A maioria (70,2%) dos padrões dos recombinantes foi similar aos padrões daquele parental ao qual ele mais se assemelhava morfologicamente auxotroficamente. Estes resultados sugerem a ocorrência de um fenômeno semelhante a transformação na parameiose
Title in English
Genetic characterization by RAPD of Trichoderma pseudokoningii recombinants
Abstract in English
For the present thesis genetic studies were carried out on the fungus Trichoderma pseudokoningii, using two independently isolated and contrasting strains,Est-3 and M. ln Est-3 five single auxotrophic mutants and four mutants with double deficiency were obtained with the use of ultraviolet light. The mutation rate was low (0.001%), suggesting that Est-3 is very resistent to ultraviolet irradiation, or may have a very efficient repair system. Two double auxotrophic mutants from Est-3 were used for protoplast fusion together with two auxotrophic mutants from the M strain, a double and a quadruple one. Recombinant colonies were obtained directly from the fusion products, however, at a low frequency (1%), indicating a reduced recombination frequency in the parameiosis between these two strains. The parental mutants and their recombinants were analyzed by RAPD, using ten primers. The parental patterns were different for all primers, suggesting a divergence between the two used strains. The majority of recombinant patterns (70.2%) was equal to the patterns of the parental to which it was more similar in morphological and auxotrophic terms. These results suggest that a phenomenon similar to transformation takes place in parameiosis.
 
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SilvaIaraFino.pdf (7.83 Mbytes)
Publishing Date
2019-12-20
 
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