title: Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application.
abstract: The cytochrome P450 gene CYP81A6 confers tolerance to bentazon and metsulfuron-methyl, two selective herbicides widely used for weed control in rice and wheat fields. Knockout mutants of CYP81A6 are highly susceptible to both herbicides. The present study aimed to characterize the CYP81A6 expression in rice. Quantitative real-time polymerase chain reaction (PCR) analyses demonstrated that foliar treatment of bentazon (500 mg/L) greatly induced expression of CYP81A6 in both wild-type (Jiazhe B) and its knockout mutant (Jiazhe mB): a 10-fold increase at 9 h before returning to basal levels at 24 h in Jiazhe B, while in the mutant the expression level rose to >20-fold at 12 h and maintained at such high level up to 24 h post exposure. In contrast, metsulfuron-methyl (500 mg/L) treatment did not affect the expression of CYP81A6 in Jiazhe B within 80 h; thereafter the expression peaked at 120 h and returned gradually to basal levels by Day 6. We suggest that a metabolite of metsulfuron-methyl, 1H-2,3-benzothiazin-4-(3H)-one-2,2-dioxide, is likely to be responsible for inducing CYP81A6 expression, rather than the metsulfuron-methyl itself. Use of a promoter-GUS reporter construct (CYP81A6Pro::GUS) demonstrated that CYP81A6 was constitutively expressed throughout the plant, with the highest expression in the upper surfaces of leaves. Subcellular localization studies in rice protoplasts showed that CYP81A6 was localized in the endoplasmic reticulum. These observations advance our understanding of CYP81A6 expression in rice, particularly its response to the two herbicides.
authors: State Key Laboratory of Rice Biology, Institute of Crop Sciences, Zhejiang University, Hangzhou 310029, China; Newcastle Institute for Research on Environment, School of Biology, University of Newcastle, Newcastle upon Tyne, NE1 7RU, UK; Jiaxing Academy of Agricultural Sciences, Jiaxing 314016, China.
authors: Hai-ping
authors: HP
authors: Lu
authors: Martin
authors: M
authors: Edwards
authors: Qi-zhao
authors: QZ
authors: Wang
authors: Hai-jun
authors: HJ
authors: Zhao
authors: Hao-wei
authors: HW
authors: Fu
authors: Jian-zhong
authors: JZ
authors: Huang
authors: Angharad
authors: A
authors: Gatehouse
authors: Qing-yao
authors: QY
authors: Shu
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000502:physiology*; D006540:Herbicides / Q000494:pharmacology*; D012275:Oryza / Q000187:drug effects / Q000201:enzymology*; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology*; D014018:Tissue Distribution
chemical_list: D006540:Herbicides; D003577:Cytochrome P-450 Enzyme System
keywords: Bentazon; CYP81A6; Expression induction; Metsulfuron-methyl; Xenobiotics
references: Plant Cell Physiol. 1996 Mar;37(2):117-22
references: 8665091
references: Annu Rev Plant Biol. 2010;61:49-64
references: 20192736
references: Nat Protoc. 2007;2(7):1565-72
references: 17585298
references: Genetics. 2001 May;158(1):41-64
references: 11333217
references: Methods. 2001 Dec;25(4):402-8
references: 11846609
references: Curr Drug Metab. 2005 Dec;6(6):545-52
references: 16379668
references: Biochim Biophys Acta. 2011 Jan;1814(1):14-8
references: 20736090
references: Plant Mol Biol. 2004 May;55(3):327-42
references: 15604685
references: J Biol Chem. 2003 Feb 28;278(9):7639-44
references: 12496246
references: Plant J. 2011 Dec;68(6):1061-9
references: 21880027
references: Plant Cell. 1991 Oct;3(10):1085-1094
references: 12324583
references: PLoS One. 2008;3(3):e1818
references: 18350155
references: Biochim Biophys Acta. 2007 Mar;1770(3):330-44
references: 16978787
references: J Exp Bot. 2013 Apr;64(7):1795-804
references: 23408830
references: Trends Plant Sci. 2007 Jan;12(1):6-13
references: 17161644
references: Nat Protoc. 2008;3(5):824-34
references: 18451790
references: Theor Appl Genet. 2002 Jul;105(1):16-22
references: 12582557
references: Pestic Biochem Physiol. 1996 May;55(1):54-63
references: 8980030
references: Plant Cell Physiol. 2012 Jun;53(6):987-1002
references: 22470056
references: Plant Mol Biol. 2006 Aug;61(6):933-43
references: 16927205