Grantee Research Project Results

2004 Progress Report: Transgenic Plants for Bioremediation of Atrazine and Related Herbicides

EPA Grant Number: R829479C019
Subproject: this is subproject number 019 , established and managed by the Center Director under grant R829479
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: Center for Air, Climate, and Energy Solutions
Center Director: Robinson, Allen
Title: Transgenic Plants for Bioremediation of Atrazine and Related Herbicides
Investigators: Sadowsky, Michael J.
Current Investigators: Sadowsky, Michael J. , Samac, Deborah A. , Vance, Carroll P. , Wackett, Lawrence P.
Institution: University of Minnesota
EPA Project Officer: Aja, Hayley
Project Period: January 1, 2004 through December 31, 2004 (Extended to December 31, 2005)
Project Period Covered by this Report: January 1, 2004 through December 31, 2005
RFA: The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program (2001) RFA Text |  Recipients Lists
Research Category: Hazardous Waste/Remediation , Targeted Research

Objective:

The objectives of this research project are to:

1. produce transgenic alfalfa plants containing a “plant expressable” atrazine chlorohydrolase gene (atzA);
2. quantify the expression of atzA, at the RNA and protein level, in the transgenic plants;
3. and evaluate the usefulness of the transgenic alfalfa plants to remove residual atrazine from herbicide-impacted soil and water.

Progress Summary:

The bacterial gene atzA encodes for atrazine chlorohydrolase, the first enzyme in a six-step bacterial pathway leading to the complete mineralization of the herbicide atrazine in gram-negative and some gram-positive soil bacteria. AtzA catalyzes the hydrolytic dechlorination and detoxification of atrazine to hydroxyatrazine, which is nonherbicidal. We investigated the potential use of transgenic plants expressing atzA to uptake, degrade, and detoxify atrazine. We subjected three plant species, Medicago sativa, Arabidopsis thaliana, and Nicotiana tabacum to Agrobacterium-mediated transformation using a bacterial atrazine chlorohydrolase gene, p-atzA, that was modified for plant codon usage. Expression of p-atzA was under control of the CsVMV promoter. Southern and Western Blot analyses and RT-PCR indicated that transgenic alfalfa, tobacco, and Arabidopsis plants maintained and expressed p-AtzA. Transgenic plants of each species grew over a wide range of atrazine concentrations, whereas control plants died at much lower concentrations. Thin layer chromatographic analyses indicated that in planta expression of p-AtzA resulted in the production of hydroxyatrazine, a nonphytotoxic compound. Moreover, hydroponically grown, transgenic, tobacco, and alfalfa lines took up atrazine from the growth medium and degraded the herbicide to hydroxyatrazine. Initial studies suggest that in planta expression of p-AtzA may be a useful trait for engineering plants that are suitable for the phytoremediation of atrazine-contaminated soils and soil water.

Future Activities:

We will continue evaluating plants for atrazine biodegradation ability under more rigorous hydroponic growth conditions and for their ability to transform atrazine to hydroxyatrazine in soils. We currently are evaluating whether monocot plant species can be transformed with p-atzA, and whether they have the ability to express p-atzA in planta to degrade atrazine to hydroxyatrazine.

Journal Articles:

No journal articles submitted with this report: View all 2 publications for this subproject

Supplemental Keywords:

atrazine, transgenic plants, transformation, hydroxyatrazine, detoxification, sustainable industry, sustainable business, waste, agricultural engineering, bioremediation, environmental engineering, geochemistry, new technology, innovative technology, bioaccumulation, biodegradation, bioenergy, bioengineering, biotechnology, phytoremediation, plant biotechnology, sustainable industry/business, environmental chemistry,, Sustainable Industry/Business, Scientific Discipline, TREATMENT/CONTROL, Waste, Agricultural Engineering, Environmental Chemistry, Treatment Technologies, Bioremediation, New/Innovative technologies, Environmental Engineering, Technology, biotechnology, phytoremediation, plant-based remediation, atrazine, remediation, biodegradation, bioacummulation, bioengineering, bioenergy, transgenic plants

Relevant Websites:

http://www.cpbr.org Exit

Progress and Final Reports:

Original Abstract

2005 Progress Report

Final

Main Center Abstract and Reports:

R829479    Center for Air, Climate, and Energy Solutions

Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R829479C001 Plant Genes and Agrobacterium T-DNA Integration
R829479C002 Designing Promoters for Precision Targeting of Gene Expression
R829479C003 aka R829479C011 Biological Effects of Epoxy Fatty Acids
R829479C004 Negative Sense Viral Vectors for Improved Expression of Foreign Genes in Insects and Plants
R829479C005 Development of Novel Plastics From Agricultural Oils
R829479C006 Conversion of Paper Sludge to Ethanol
R829479C007 Enhanced Production of Biodegradable Plastics in Plants
R829479C008 Engineering Design of Stable Immobilized Enzymes for the Hydrolysis and Transesterification of Triglycerides
R829479C009 Discovery and Evaluation of SNP Variation in Resistance-Gene Analogs and Other Candidate Genes in Cotton
R829479C010 Woody Biomass Crops for Bioremediating Hydrocarbons and Metals
R829479C011 Biological Effects of Epoxy Fatty Acids
R829479C012 High Strength Degradable Plastics From Starch and Poly(lactic acid)
R829479C013 Development of Herbicide-Tolerant Energy and Biomass Crops
R829479C014 Identification of Receptors of Bacillus Thuringiensis Toxins in Midguts of the European Corn Borer
R829479C015 Coordinated Expression of Multiple Anti-Pest Proteins
R829479C016 A Novel Fermentation Process for Butyric Acid and Butanol Production from Plant Biomass
R829479C017 Molecular Improvement of an Environmentally Friendly Turfgrass
R829479C018 Woody Biomass Crops for Bioremediating Hydrocarbons and Metals. II.
R829479C019 Transgenic Plants for Bioremediation of Atrazine and Related Herbicides
R829479C020 Root Exudate Biostimulation for Polyaromatic Hydrocarbon Phytoremediation
R829479C021 Phytoremediation of Heavy Metal Contamination by Metallohistins, a New Class of Plant Metal-Binding Proteins
R829479C022 Development of Herbicide-Tolerant Energy and Biomass Crops
R829479C023 A Novel Fermentation Process for Butyric Acid and Butanol Production from Plant Biomass
R829479C024 Development of Vectors for the Stoichiometric Accumulation of Multiple Proteins in Transgenic Crops
R829479C025 Chemical Induction of Disease Resistance in Trees
R829479C026 Development of Herbicide-Tolerant Hardwoods
R829479C027 Environmentally Superior Soybean Genome Development
R829479C028 Development of Efficient Methods for the Genetic Transformation of Willow and Cottonwood for Increased Remediation of Pollutants
R829479C029 Development of Tightly Regulated Ecdysone Receptor-Based Gene Switches for Use in Agriculture
R829479C030 Engineered Plant Virus Proteins for Biotechnology