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Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application

Lookup NU author(s): Dr Martin Edwards, Professor Angharad MR Gatehouse

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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.


Publication metadata

Author(s): Lu H, Edwards M, Wang Q, Zhao H, Fu H, Huang J, Gatehouse A, Shu Q

Publication type: Article

Publication status: Published

Journal: Journal of Zhejiang University-Science B

Year: 2015

Volume: 16

Issue: 2

Pages: 113-122

Print publication date: 01/02/2015

Online publication date: 05/03/2015

Acceptance date: 12/08/2014

ISSN (print): 1673-1581

ISSN (electronic): 1862-1783

Publisher: Springer

URL: http://dx.doi.org/10.1631/jzus.B1400168

DOI: 10.1631/jzus.B1400168


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