Biodiversidade do gênero Trichoderma (Hypocreales - fungi) mediante técnicas moleculares e análise ecofisiográfica
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Data
2005-01-28
Autores
Corabi-Adell, Carlo [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A diversidade do gênero Trichoderma no estado de São Paulo (BRASIL) foi analisada sob diferentes aspectos. Foi estabelecida uma coleção de 539 linhagens obtidas a partir de 52 pontos de coleta, distribuídos pelo estado, e uma coleção de referência com 30 linhagens. Durante o período de trabalho avaliou-se a viabilidade das culturas de acordo com os métodos de preservação utilizados Castellani e câmara fria. As avaliações do potencial de biocontrole pelas técnicas de pareamento e difusão de metabólitos inibidores não apresentaram uma correspondência clara com os testes em casa de vegetação. Para as observações microscópicas de culturas de lâminas foi desenvolvida uma técnica que permitiu visualizar mais facilmente o sistema de ramificação dos conidióforos. A avaliação celulolítica dos isolados permitiu identificar linhagens com atividade superior a da linhagem de referência, indicando alto potencial biotecnológico. A análise molecular dos isolados a partir do seqüenciamento da região ITS1-5,8S-ITS2 mostrou a ocorrência de 17 espécies distribuídas no estado, sobretudo nas áreas de mata e capoeira. As espécies mais freqüentes, T. harzianum, T. spirale e T. koningii ocorreram em quase todos os ecossistemas avaliados, enquanto a ocorrência da seção Longibrachiatum foi pouco expressiva. A técnica de RAPD se mostrou eficiente na identificação de linhagens redundantes indicando seu potencial de uso durante procedimentos de triagem. A aplicação de métodos de taxonomia numérica permitiu gerar fenogramas e gráficos multidimensionais coerentes com os dados morfológicos e moleculares em diversos casos. É sugerida a aplicação de métodos topológicos para a melhor descrição do sistema de ramificação e diferenciação de hifas e da teoria de sistemas não-lineares para a compreensão do seu comportamento morfológico in e ex vitro...
The diversity of the genus Trichoderma in São Paulo State (BRAZIL) was analised under different views. A collection of 539 strains was isolated from 52 different areas and a reference collection of 30 strains was established from different brazilian culture collections. The preservation techniques of Castellani and cold storage were evaluated during the course of the study. The biocontrol potential was evaluated through pairing cultures methodology and inhibitory metabolites production. There was no clear evidence of correspondence between the data obtained in vitro and under greenhouse conditions. For microscopic observation a new technique of slide culture was developed in order to visualise the branching system easily. Cellulolytic activitiy assay identified several high-producing cellulase strains comparing to the reference QM9414 indicating a biotechnological potential. The molecular analysis from the sequencing of ITS1-5,8S-ITS2 region indicated the ocurrency of 17 different species all over the state with predominance in forest and 'capoeira' ecossystems. The most frequently species were T.harzianum, T.spirale and T.koningii, ocurring in almost all environmental conditions sampled, while the presence of section Longibrachiatum was inexpressive. The RAPD technique in separate redundants strains was demonstrated indicating its potential for using in screening procedures. Numerical taxonomy methods produced phenograms and multidimensional graphics consistent with molecular and morphological data in some cases. The use of topological methods in describing the branching system and the non-linear system theory in the comprehension of its morphological behaviour in and ex vitro (anamorphic, teleomorphic and synanomorphic states) is suggested. For the first time the total evidence analysis was applied to the genus, showing consistent results with those reported in literature.
The diversity of the genus Trichoderma in São Paulo State (BRAZIL) was analised under different views. A collection of 539 strains was isolated from 52 different areas and a reference collection of 30 strains was established from different brazilian culture collections. The preservation techniques of Castellani and cold storage were evaluated during the course of the study. The biocontrol potential was evaluated through pairing cultures methodology and inhibitory metabolites production. There was no clear evidence of correspondence between the data obtained in vitro and under greenhouse conditions. For microscopic observation a new technique of slide culture was developed in order to visualise the branching system easily. Cellulolytic activitiy assay identified several high-producing cellulase strains comparing to the reference QM9414 indicating a biotechnological potential. The molecular analysis from the sequencing of ITS1-5,8S-ITS2 region indicated the ocurrency of 17 different species all over the state with predominance in forest and 'capoeira' ecossystems. The most frequently species were T.harzianum, T.spirale and T.koningii, ocurring in almost all environmental conditions sampled, while the presence of section Longibrachiatum was inexpressive. The RAPD technique in separate redundants strains was demonstrated indicating its potential for using in screening procedures. Numerical taxonomy methods produced phenograms and multidimensional graphics consistent with molecular and morphological data in some cases. The use of topological methods in describing the branching system and the non-linear system theory in the comprehension of its morphological behaviour in and ex vitro (anamorphic, teleomorphic and synanomorphic states) is suggested. For the first time the total evidence analysis was applied to the genus, showing consistent results with those reported in literature.
Descrição
Palavras-chave
Microorganismos, Celulase, Filogenia, RAPD, ITS, Taxonomia, Taxonomy, Cellulase
Como citar
CORABI-ADELL, Carlo. Biodiversidade do gênero Trichoderma (Hypocreales - fungi) mediante técnicas moleculares e análise ecofisiográfica. 2004. xviii, 202 f. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Biociências, 2004.