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Project 5: Phytorermediation of Pollutants Using Transgenic Plants

项目5：利用转基因植物修复污染物

基本信息

批准号：
8254488
负责人：
Stuart E Strand
金额：
$ 31.87万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-07 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8254488
关键词：
Air Amendment Beds Benzene Biological Assay Biological Markers Candidate Disease Gene Carbon Tetrachloride Categories Cell Culture Techniques Chloroform Chlorpyrifos Clinical Collaborations Cytochrome P-450 CYP2E1 Diazinon Environment Equilibrium Excision Exposure to Flowers Fractionation Funding Gene Expression Profile Genes Genetic Genetically Modified Plants Health Hepatic Home environment Human Irrigation Isotope Labeling Left Mammals Measurement Metabolism Methods Microarray Analysis Mixed Function Oxygenases Motivation National Institute of Environmental Health Sciences Neurotoxins Organophosphates Organophosphorus Compounds Oryctolagus cuniculus Parathion Pathway interactions Pesticides Phenotype Pheretima sieboldi Plant Genes Plant Leaves Plant Roots Plants Poison Predisposition RNA Interference Reverse Transcriptase Polymerase Chain Reaction Risk Safety Sarin Site Soil Soman Stomas Superfund Surface Testing Therapeutic Uses Tissues Toxic effect Toxin Transgenes Transgenic Organisms Transgenic Plants Trees Trichloroethylene Water Work Yeasts aqueous aryldialkylphosphatase dechlorination exposed human population fusicoccin nerve agent neurotoxic overexpression oxidation plant growth/development pollutant prevent remediation research study response uptake volatile organic compound

项目摘要

Exposure to toxic volatile organic compounds (VOCs) continues to pose a significant risk to human health. VOCs such as chloroform, carbon tetrachloride (CT) and trichloroethylene (TCE), along with benzene and other single-ring aromatics are among the most common contaminants in water and air. These compounds share a susceptibility to oxidation by the mammalian hepatic cytochrome P450 2E1. In past NIEHS-funded work at UW, we have demonstrated that TCE and CT are oxidized by axenic poplar cell cultures using a pathway similar to that in mammals, and that wild-type poplar are able to take up and degrade TCE and CT. Our work has led to the realization that TCE and CT uptake by wild-type plants is too weak to significantly reduce their concentration in root zone water, providing motivation for the development of plants with increased degradative activity toward VOCs. Toward this end we have expressed mammalian cytochrome P450 2E1 (CYP2E1) in plants, achieving orders of magnitude greater oxidation of TCE in transgenic poplar. We are presently engaged in testing TCE degradation by wild-type and CYP2E1 transgenic poplar at field scale in a test bed facility capable of mass balance measurements. In this application we propose the following specific aims: compare the abilities of wild-type and existing CYP2E1 transgenic poplar lines to degrade TCE, CT, and PCE in field tests; identify genes that are up-regulated by the presence of pollutants; clone and analyze the plant genes involved in the degradation of TCE and its metabolites; determine the metabolites of TCE and benzene in CYP2E1 transgenic poplar; increase the uptake of VOCs from the air by increasing the number of open stomata on leaf surfaces; express the organophosphorus hydroxylase, paraoxonase PON1, in plants for the degradation of organophosphate neurotoxins; and determine the ecotoxicity to earthworms of root materials, leaf litter and rhizosphere soil of CYP2E1 transgenic plants exposed to VOCs.

接触有毒挥发性有机化合物（VOC）继续对人类健康构成重大风险。挥发性有机化合物如氯仿、四氯化碳（CT）和三氯乙烯（TCE），沿着苯和其他单环芳烃是水和空气中最常见的污染物。这些化合物对哺乳动物肝细胞色素P450 2 E1的氧化具有敏感性。在UW过去NIEHS资助的工作中，我们已经证明了TCE和CT被纯白杨细胞培养物氧化，使用与哺乳动物相似的途径，并且野生型白杨能够吸收和降解TCE和CT。我们的工作使人们认识到，野生型植物对TCE和CT的吸收太弱，无法显著降低其在根区水中的浓度，这为开发具有增加的VOCs降解活性的植物提供了动力。为此，我们在植物中表达了哺乳动物细胞色素P450 2 E1（CYP 2 E1），在转基因白杨中实现了更大数量级的TCE氧化。目前我们正在进行野生型杨树和转CYP 2 E1基因白杨对三氯乙烯的田间降解试验能够进行质量平衡测量的试验台设施中的天平。本研究的主要目的是：通过田间试验比较转CYP 2 E1基因白杨和野生型杨树对三氯乙烯（TCE）、四氯化碳（CT）和四氯乙烯（PCE）的降解能力，鉴定受污染物影响而表达上调的基因，克隆和分析与三氯乙烯及其代谢产物降解相关的植物基因，确定转CYP 2 E1基因杨树中三氯乙烯和苯的代谢产物，并对转CYP 2 E1基因杨树中三氯乙烯和苯的代谢产物进行分析。通过增加叶片表面开放气孔的数量来增加从空气中吸收VOCs;在植物中表达有机磷羟化酶，对氧磷酶PON 1，用于降解有机磷神经毒素;以及确定暴露于VOCs的CYP 2 E1转基因植物的根材料、叶凋落物和根际土壤对蚯蚓的生态毒性。