title: A collection of cytochrome P450 monooxygenase genes involved in modification and detoxification of herbicide atrazine in rice (Oryza sativa) plants.
abstract: Plant cytochrome P450 monooxygenases constitute one of the largest families of protein genes involved in plant growth, development and acclimation to biotic and abiotic stresses. However, whether these genes respond to organic toxic compounds and their biological functions for detoxifying toxic compounds such as herbicides in rice are poorly understood. The present study identified 201 genes encoding cytochrome P450s from an atrazine-exposed rice transcriptome through high-throughput sequencing. Of these, 69 cytochrome P450 genes were validated by microarray and some of them were confirmed by real time PCR. Activities of NADPH-cytochrome P450 reductase (CPR) and p-nitroanisole O-demethylase (PNOD) related to toxicity were determined and significantly induced by atrazine exposure. To dissect the mechanism underlying atrazine modification and detoxification by P450, metabolites (or derivatives) of atrazine in plants were analyzed by ultra performance liquid chromatography mass spectrometry (UPLC/MS). Major metabolites comprised desmethylatrazine (DMA), desethylatrazine (DEA), desisopropylatrazine (DIA), hydroxyatrazine (HA), hydroxyethylatrazine (HEA) and hydroxyisopropylatrazine (HIA). All of them were chemically modified by P450s. Furthermore, two specific inhibitors of piperonyl butoxide (PBO) and malathion (MAL) were used to assess the correlation between the P450s activity and rice responses including accumulation of atrazine in tissues, shoot and root growth and detoxification.
authors.affiliation: Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China; Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Rong Tan
authors.affiliation: Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China; Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen Lu
authors.affiliation: Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China; Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Jing Zhang
authors.affiliation: Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Luo
authors.affiliation: Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: hongyang@njau.edu.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Yang
mesh_terms: D001280:Atrazine / Q000633:toxicity*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004781:Environmental Exposure; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D006540:Herbicides / Q000633:toxicity*; D008658:Inactivation, Metabolic; D046228:Microarray Analysis; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D012275:Oryza / Q000187:drug effects / Q000201:enzymology*; D010090:Oxidoreductases, O-Demethylating / Q000378:metabolism; D010882:Piperonyl Butoxide; D018517:Plant Roots / Q000187:drug effects / Q000378:metabolism; D059467:Transcriptome / Q000187:drug effects
chemical_list: D006540:Herbicides; D003577:Cytochrome P-450 Enzyme System; C026048:4-nitroanisole O-demethylase; D010090:Oxidoreductases, O-Demethylating; D009251:NADPH-Ferrihemoprotein Reductase; D010882:Piperonyl Butoxide; D001280:Atrazine
keywords: Atrazine; Deep-sequencing; Degradation; Microarray; P450s; Rice