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Bioresour Technol ; 101(8): 2888-91, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19897358

RESUMO

The species of the genus Trichoderma are used successfully as biocontrol agents against a wide range of phytopathogenic fungi. Among them, Trichoderma harzianum is especially effective. However, to develop more effective fungal biocontrol strategies in organic substrates and soil, tools for monitoring the control agents are required. Real-time PCR is potentially an effective tool for the quantification of fungi in environmental samples. The aim of this study consisted of the development and application of a real-time PCR-based method to the quantification of T. harzianum, and the extrapolation of these data to fungal biomass values. A set of primers and a TaqMan probe for the ITS region of the fungal genome were designed and tested, and amplification was correlated to biomass measurements obtained with optical microscopy and image analysis, of the hyphal length of the mycelium of the colony. A correlation of 0.76 between ITS copies and biomass was obtained. The extrapolation of the quantity of ITS copies, calculated based on real-time PCR data, into quantities of fungal biomass provides potentially a more accurate value of the quantity of soil fungi.


Assuntos
Contagem de Colônia Microbiana/métodos , Controle Biológico de Vetores/métodos , Trichoderma/crescimento & desenvolvimento , Trichoderma/genética , Sequência de Bases , Biomassa , Primers do DNA/genética , DNA Espaçador Ribossômico/genética , Hifas/crescimento & desenvolvimento , Dados de Sequência Molecular , Densidade Demográfica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Taq Polimerase
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