Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR

Detalhes bibliográficos
Autor(a) principal: Queiroz, Paulo Roberto
Data de Publicação: 2004
Outros Autores: Valadares-Inglis, Maria Cléria, Inglis, Peter Ward
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Pesquisa Agropecuária Brasileira (Online)
Texto Completo: https://seer.sct.embrapa.br/index.php/pab/article/view/6787
Resumo: The objective of this work was to evaluate the survival of two Trichoderma harzianum cotransformants, 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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spelling Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCRSobrevivência em solo e detecção de co-transformantes de Trichoderma harzianum por PCR "nested"biological control; transformation; egfp; benomyl resistancecontrole biológico; transformação; egfp; resistência a benomilThe objective of this work was to evaluate the survival of two Trichoderma harzianum cotransformants, 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.O objetivo deste trabalho foi avaliar a sobrevivência de dois co-transformantes de Trichoderma harzianum, TE 10 e TE 41, expressando o gene da proteína de fluorescência verde (egfp) e resistência a benomil, por um período de quatro semanas em microcosmo de solo, sob condições controladas. Foi utilizado um meio seletivo para detecção de material fúngico viável, o qual foi confirmado por observação quanto ao fenótipo de fluorescência em microscópio de epifluorescência direta. O fungo transformado foi detectado por PCR "nested", utilizando-se dois pares de primers específicos para o gene egfp. Foram utilizados meios líquidos enriquecidos no cultivo de amostras de solo, permitindo uma detecção consistente de co-transformantes de T. harzianum, uma vez que não foi possível a detecção do gene egfp por PCR de amostras de DNA extraídas diretamente de solo.Pesquisa Agropecuaria BrasileiraPesquisa Agropecuária BrasileiraQueiroz, Paulo RobertoValadares-Inglis, Maria ClériaInglis, Peter Ward2004-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.sct.embrapa.br/index.php/pab/article/view/6787Pesquisa Agropecuaria Brasileira; v.39, n.4, abr. 2004; 403-405Pesquisa Agropecuária Brasileira; v.39, n.4, abr. 2004; 403-4051678-39210100-104xreponame:Pesquisa Agropecuária Brasileira (Online)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPAenghttps://seer.sct.embrapa.br/index.php/pab/article/view/6787/3843info:eu-repo/semantics/openAccess2014-07-17T17:23:36Zoai:ojs.seer.sct.embrapa.br:article/6787Revistahttp://seer.sct.embrapa.br/index.php/pabPRIhttps://old.scielo.br/oai/scielo-oai.phppab@sct.embrapa.br || sct.pab@embrapa.br1678-39210100-204Xopendoar:2014-07-17T17:23:36Pesquisa Agropecuária Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false
dc.title.none.fl_str_mv Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR
Sobrevivência em solo e detecção de co-transformantes de Trichoderma harzianum por PCR "nested"
title Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR
spellingShingle Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR
Queiroz, Paulo Roberto
biological control; transformation; egfp; benomyl resistance
controle biológico; transformação; egfp; resistência a benomil
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
author Queiroz, Paulo Roberto
author_facet Queiroz, Paulo Roberto
Valadares-Inglis, Maria Cléria
Inglis, Peter Ward
author_role author
author2 Valadares-Inglis, Maria Cléria
Inglis, Peter Ward
author2_role author
author
dc.contributor.none.fl_str_mv

dc.contributor.author.fl_str_mv Queiroz, Paulo Roberto
Valadares-Inglis, Maria Cléria
Inglis, Peter Ward
dc.subject.por.fl_str_mv biological control; transformation; egfp; benomyl resistance
controle biológico; transformação; egfp; resistência a benomil
topic biological control; transformation; egfp; benomyl resistance
controle biológico; transformação; egfp; resistência a benomil
description The objective of this work was to evaluate the survival of two Trichoderma harzianum cotransformants, 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.
publishDate 2004
dc.date.none.fl_str_mv 2004-04-01
dc.type.none.fl_str_mv
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://seer.sct.embrapa.br/index.php/pab/article/view/6787
url https://seer.sct.embrapa.br/index.php/pab/article/view/6787
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://seer.sct.embrapa.br/index.php/pab/article/view/6787/3843
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Pesquisa Agropecuaria Brasileira
Pesquisa Agropecuária Brasileira
publisher.none.fl_str_mv Pesquisa Agropecuaria Brasileira
Pesquisa Agropecuária Brasileira
dc.source.none.fl_str_mv Pesquisa Agropecuaria Brasileira; v.39, n.4, abr. 2004; 403-405
Pesquisa Agropecuária Brasileira; v.39, n.4, abr. 2004; 403-405
1678-3921
0100-104x
reponame:Pesquisa Agropecuária Brasileira (Online)
instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
instacron:EMBRAPA
instname_str Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
instacron_str EMBRAPA
institution EMBRAPA
reponame_str Pesquisa Agropecuária Brasileira (Online)
collection Pesquisa Agropecuária Brasileira (Online)
repository.name.fl_str_mv Pesquisa Agropecuária Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
repository.mail.fl_str_mv pab@sct.embrapa.br || sct.pab@embrapa.br
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