2004 Progress Report: Enhanced Production of Biodegradable Plastics in PlantsEPA Grant Number: R829479C007
Subproject: this is subproject number 007 , 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: The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program
Center Director: Schumacher, Dorin
Title: Enhanced Production of Biodegradable Plastics in Plants
Investigators: Mooney, Brian Patrick
Institution: University of Missouri - Columbia
EPA Project Officer: Lasat, Mitch
Project Period: June 1, 2003 through December 31, 2006 (Extended to December 31, 2007)
Project Period Covered by this Report: June 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
The objectives of this research project are to:
- confirm the plastid-targeting of the chimeric Arabidopsis thaliana branched-chain dehydrogenase alpha and beta subunits (AtBCE1 and AtBCE1ß);
- confirm the protein-protein association of the chimeric AtBCE1 enzyme (composed of 2 and 2ß subunits) with the endogenous plastid dihydrolipoyl acteyltransferase (E2) enzyme;
- and generate transgenic plants expressing plastid-targeted AtBCE1 that produce elevated amounts of propionyl-CoA.
Objective 1 (completed)
Two constructs were generated from the original cDNAs encoding the A. thaliana branched-chain dehydrogenase, which consists of and ß subunits. The mitochondrial targeting sequences of both subunits were replaced by plastid transit peptides (derived from the small subunit of ribulose bisphosphate carboxylase/oxygenase). The redirection of targeting to the plastid was confirmed by in vitro import assays using isolated pea chloroplasts.
Objective 2 (ongoing)
To date, we have only attempted chloroplast import separately for each subunit. We will conduct coimports of the two subunits into isolated pea chloroplasts. We then will confirm the association of the and ß subunits of the branched-chain dehydrogenase with the endogenous plastid pyruvate dehydrogenase complex E2 enzyme by immunoprecipitation with antibodies specific to the E2 protein.
Objective 3 (ongoing)
The cDNAs encoding the chimeric subunits were inserted into separate plant transformation vectors pBI121 and pCambia1201. A. thaliana plants were transformed separately with constructs encoding the individual subunits. We have just harvested seeds from these transformed plants and are in the process of identifying plants that are expressing the subunits. We also are pursuing a cotransformation strategy. We have identified colonies of Agrobacteria that contain both constructs and are waiting for the A. thaliana plants to reach the flowering stage for transformation.
A news item in Mizzou Weekly, a University of Missouri–Columbia campus publication, described the project and its potential benefits for Missouri farmers, once proof of concept has been established. This article was reprinted in the MU Alumni magazine, in the annual report for the University of Missouri system, and in Jarman’s Weekly Agronomy News, a University of Missouri Extension publication. A news brief item in Life Sciences Update, a University of Missouri–Columbia campus publication, also was published describing the project outline and impacts.
Seeds will be collected from the transgenic plants recently generated. These seeds will be germinated in the presence of a chemical that inhibits growth of the plant unless the gene of interest has been incorporated into the plant genome (i.e., the transformation has been successful). Once we have confirmed that the transgenic plants contain the two genes of interest, they will be characterized for the expression of the genes and the levels of the biodegradable plastic precursor chemicals.
Journal Articles:No journal articles submitted with this report: View all 6 publications for this subproject
Supplemental Keywords:plastic, plastid, Arabidopsis thaliana, branched chain, dehydrogenase, AtBCE1, AtBCE1ß, enzyme, transgenic, propionyl-CoA, sustainable industry/business, treatment/control, agricultural engineering, environmental engineering, genetics, geochemistry, new/innovative technologies, technology, biodegradable plastics, bioenergy, bioengineering, biotechnology, novel plastics, plant biotechnology,, Scientific Discipline, TREATMENT/CONTROL, Sustainable Industry/Business, Genetics, Geochemistry, Technology, New/Innovative technologies, Environmental Engineering, Agricultural Engineering, bioengineering, biodegradable plastics, plant genes, biotechnology, plant biotechnology, novel plastics
Progress and Final Reports:Original Abstract
Main Center Abstract and Reports:R829479 The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program
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