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Expression of Anti-Atrazine scFv and Atrazine Chlorohydrolase TrzN in planta for Potential Phytoremediation of Atrazine Contamination

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Title: Expression of Anti-Atrazine scFv and Atrazine Chlorohydrolase TrzN in planta for Potential Phytoremediation of Atrazine Contamination
Author: Leelachao, Siripen
Department: School of Environmental Sciences
Program: Toxicology
Advisor: Hall, ChristopherSolomon, Keith
Abstract: Atrazine is extensively used in agricultural production to improve crop yield throughout North America. Due to contamination of surface water with atrazine, which is an environmental concern, tools for mitigating such contamination are needed. In this study, the potential use of genetically modified plants for remediation of atrazine contamination was investigated. Plants were genetically modified to express proteins of interest using Agrobacterium-mediated transformation. First, an anti-atrazine single chain variable fragment (scFv) was expressed in Lemna minor. In a hydroponic system, transgenic L. minor expressing the scFv showed higher tolerance to atrazine than wild-type plants. Absorption of 14C-atrazine in transgenic L. minor was greater than wild-type L. minor. Metabolites of atrazine were not found in transgenic L. minor. Second, L. minor was transformed to express atrazine chlorohydrolase (TrzN), but the protein could not be detected. Resistance to atrazine was found in the transgenic plants at low concentrations. Finally, the gene of atrazine chlorohydrolase TrzN was tested in Nicotiana Benthamiana by agroinfiltration. Transient Expression of atrazine chlorohydrolase TrzN and tolerance to atrazine in transgenic N. benthamiana were evaluated through agroinfiltration followed by simulation of a field spray of atrazine. Tolerance to atrazine was induced by atrazine chlorohydrolase TrzN. The atrazine chlorohydrolase TrzN was produced in stable transgenic N. benthamiana. The protein was bound to cell membranes. The transgenic N. benthamiana were tolerant to atrazine when grown in both a hydroponic system and in soil with application of spray to the plants.
URI: http://hdl.handle.net/10214/8745
Date: 2015-04


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