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"

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ID: 257457
2004
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Ranked #20 of 193 articles by views in proc - the fourth ieee workshop on software technol for future embedded and ubiquitous syst, seus 2006 andthe second int workshop on collaborative comput, integr, and assur, wccia 2006

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Abstract
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.
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.
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queiroz2004pesquisasurvival Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors ;Paulo Roberto Queiroz;Maria Cléria Valadares-Inglis;Peter Ward Inglis
Journal proc - the fourth ieee workshop on software technol for future embedded and ubiquitous syst, seus 2006 andthe second int workshop on collaborative comput, integr, and assur, wccia 2006
Year 2004
DOI
10.1590/S0100-204X2004000400015
URL
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2004000400015
Keywords
biological control transformation egfp controle biológico benomyl resistanceagriculture (general)

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