Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR
The objective of this work was to evaluate the survival of two Trichoderma harzianum co-transformants, TE 10 and TE 41, carrying genes for green fluorescent protein (egfp) and for resistance to benomyl, during four weeks in a contained soil microcosm. Selective culture media were used to detect viable fungal material, whose identity was confirmed by the observation of the fluorescent phenotype by direct epifluorence microscopy. PCR using two nested primer pairs specific to the egfp gene was also used to detect the transformed fungi. Although it was not possible to reliably detect the egfp gene directly from soil extracts, an enrichment step involving selective culture of soil samples in liquid medium prior to DNA extraction enabled the consistent detection of the T. harzianum co-transformants by nested PCR for the duration of the incubation period.
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Embrapa Secretaria de Pesquisa e Desenvolvimento
2004
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oai:scielo:S0100-204X20040004000152004-06-24Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCRQueiroz,Paulo RobertoValadares-Inglis,Maria ClériaInglis,Peter Ward biological control transformation egfp benomyl resistance The objective of this work was to evaluate the survival of two Trichoderma harzianum co-transformants, TE 10 and TE 41, carrying genes for green fluorescent protein (egfp) and for resistance to benomyl, during four weeks in a contained soil microcosm. Selective culture media were used to detect viable fungal material, whose identity was confirmed by the observation of the fluorescent phenotype by direct epifluorence microscopy. PCR using two nested primer pairs specific to the egfp gene was also used to detect the transformed fungi. Although it was not possible to reliably detect the egfp gene directly from soil extracts, an enrichment step involving selective culture of soil samples in liquid medium prior to DNA extraction enabled the consistent detection of the T. harzianum co-transformants by nested PCR for the duration of the incubation period.info:eu-repo/semantics/openAccessEmbrapa Secretaria de Pesquisa e DesenvolvimentoPesquisa Agropecuária BrasileiraPesquisa Agropecuária Brasileira v.39 n.4 20042004-04-01info:eu-repo/semantics/reporttext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2004000400015en10.1590/S0100-204X2004000400015 |
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Queiroz,Paulo Roberto Valadares-Inglis,Maria Cléria Inglis,Peter Ward |
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Queiroz,Paulo Roberto Valadares-Inglis,Maria Cléria Inglis,Peter Ward Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR |
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Queiroz,Paulo Roberto Valadares-Inglis,Maria Cléria Inglis,Peter Ward |
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Queiroz,Paulo Roberto |
title |
Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR |
title_short |
Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR |
title_full |
Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR |
title_fullStr |
Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR |
title_full_unstemmed |
Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR |
title_sort |
survival in soil and detection of co-transformed trichoderma harzianum by nested pcr |
description |
The objective of this work was to evaluate the survival of two Trichoderma harzianum co-transformants, TE 10 and TE 41, carrying genes for green fluorescent protein (egfp) and for resistance to benomyl, during four weeks in a contained soil microcosm. Selective culture media were used to detect viable fungal material, whose identity was confirmed by the observation of the fluorescent phenotype by direct epifluorence microscopy. PCR using two nested primer pairs specific to the egfp gene was also used to detect the transformed fungi. Although it was not possible to reliably detect the egfp gene directly from soil extracts, an enrichment step involving selective culture of soil samples in liquid medium prior to DNA extraction enabled the consistent detection of the T. harzianum co-transformants by nested PCR for the duration of the incubation period. |
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Embrapa Secretaria de Pesquisa e Desenvolvimento |
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2004 |
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http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2004000400015 |
work_keys_str_mv |
AT queirozpauloroberto survivalinsoilanddetectionofcotransformedtrichodermaharzianumbynestedpcr AT valadaresinglismariacleria survivalinsoilanddetectionofcotransformedtrichodermaharzianumbynestedpcr AT inglispeterward survivalinsoilanddetectionofcotransformedtrichodermaharzianumbynestedpcr |
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1756385654742188032 |