Full Text:   <1800>

CLC number: S482.4

On-line Access: 2015-02-02

Received: 2014-04-30

Revision Accepted: 2014-08-12

Crosschecked: 2015-01-08

Cited: 6

Clicked: 3068

Citations:  Bibtex RefMan EndNote GB/T7714


Hai-ping Lu


-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.2 P.113-122


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

Author(s):  Hai-ping Lu, Martin Edwards, Qi-zhao Wang, Hai-jun Zhao, Hao-wei Fu, Jian-zhong Huang, Angharad Gatehouse, Qing-yao Shu

Affiliation(s):  State Key Laboratory of Rice Biology, Institute of Crop Sciences, Zhejiang University, Hangzhou 310029, China; more

Corresponding email(s):   qyshu@zju.edu.cn

Key Words:  CYP81A6, Bentazon, Metsulfuron-methyl, Expression induction, Xenobiotics

Hai-ping Lu, Martin Edwards, Qi-zhao Wang, Hai-jun Zhao, Hao-wei Fu, Jian-zhong Huang, Angharad Gatehouse, Qing-yao Shu. Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application[J]. Journal of Zhejiang University Science B, 2015, 16(2): 113-122.

@article{title="Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application",
author="Hai-ping Lu, Martin Edwards, Qi-zhao Wang, Hai-jun Zhao, Hao-wei Fu, Jian-zhong Huang, Angharad Gatehouse, Qing-yao Shu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application
%A Hai-ping Lu
%A Martin Edwards
%A Qi-zhao Wang
%A Hai-jun Zhao
%A Hao-wei Fu
%A Jian-zhong Huang
%A Angharad Gatehouse
%A Qing-yao Shu
%J Journal of Zhejiang University SCIENCE B
%V 16
%N 2
%P 113-122
%@ 1673-1581
%D 2015
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1400168

T1 - Expression of cytochrome P450 CYP81A6 in rice: tissue specificity, protein subcellular localization, and response to herbicide application
A1 - Hai-ping Lu
A1 - Martin Edwards
A1 - Qi-zhao Wang
A1 - Hai-jun Zhao
A1 - Hao-wei Fu
A1 - Jian-zhong Huang
A1 - Angharad Gatehouse
A1 - Qing-yao Shu
J0 - Journal of Zhejiang University Science B
VL - 16
IS - 2
SP - 113
EP - 122
%@ 1673-1581
Y1 - 2015
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1400168

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.




Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Ahmad, P., Sarwat, M., Sharma, S., 2008. Reactive oxygen species, antioxidant and signaling in plants. J. Plant Biol., 51(3):167-173.

[2]Courcelle, J., Khodursky, A., Hanawalt, P.C., 2001. Comparative gene expression profiles following UV exposure in wild-type and SOS-deficient Escherichia coli. Genetics, 158(1):41-64.

[3]Deng, F., Hatzios, K.K., 2003. Characterization of cytochrome P450-mediated bensulfuron-methyl O-demethylation in rice. Pestic. Biochem. Physiol., 74(2):102-115.

[4]Dvorak, Z., Ulrichova, J., Modriansky, M., 2005. Role of microtubules network in CYP genes expression. Curr. Drug Metab., 6(6):545-552.

[5]Edwards, R., Dixon, D.P., Cummins, I., et al., 2011. New perspectives on the metabolism and detoxification of synthetic compounds in plants. In: Schröder, P., Collins, C.D. (Eds.), Organic Xenobiotics and Plants, Vol 8. Springer Netherlands, p.125-148.

[6]Fu, H.W., 2004. Hybrid rice Jiayou 99 with fine quality and super-yield. J. Anhui Agric. Sci., 32(3):429-432 (in Chinese).

[7]Gianessi, L.P., Reigner, N.P., 2007. The value of herbicides in US crop production. Weed Technol., 21(2):559-566.

[8]Glover, D.G., Schapaugh, W.T.Jr., 2002. Inheritance of resistance to pendimethalin herbicide induced stem damage in soybean. Euphytica, 125(3):433-437.

[9]Green, J.M., 2009. Evolution of glyphosate-resistance crop technology. Weed Sci., 57(1):108-117.

[10]Han, Y.C., Wang, C.Y., 2002. Physiological basis of bentazon tolerance in rice (Oryza sativa L.) lines. Weed Biol. Manag., 2(4):186-193.

[11]Hannemann, F., Bichet, A., Ewen, K.M., et al., 2007. Cytochrome P450 systems—biological variations of electron transport chains. Biochim. Biophys. Acta, 1770(3):330-344.

[12]Hiei, Y., Komari, T., 2008. Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed. Nat. Protoc., 3(5):824-834.

[13]Konishi, T., Shinohara, K., Yamada, K., et al., 1996. Acetyl-CoA carboxylase in higher plants: most plants other than Gramineae have both the prokaryotic and the eukaryotic forms of enzyme. Plant Cell Physiol., 37(2):117-122.

[14]Lin, C.Y., Fang, J., Xu, X.L., et al., 2008. A built-in strategy for containment of transgenic plants: creation of selectively terminable transgenic rice. PLoS ONE, 3(3):e1818.

[15]Liu, C., Liu, S.Q., Wang, F., et al., 2012. Expression of a rice CYP81A6 gene confers tolerance to bentazon and sulfonylurea herbicides in both Arabidopsis and tobacco. Plant Cell Tiss. Organ Cult., 109(3):419-428.

[16]Liu, M.Y., Chen, W.W., Xu, J.M., et al., 2013. The role of VuMATE1 expression in aluminum-inducible citrate secretion in rice bean (Vigna umbellata) roots. J. Exp. Bot., 64(7):1795-1804.

[17]Livak, K.J., Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT. Methods, 25(4):402-408.

[18]Lv, M.Z., Chao, D.Y., Shan, J.X., et al., 2012. Rice carotenoid β-ring hydroxylase CYP97A4 is involved in lutein biosynthesis. Plant Cell Physiol., 53(6):987-1002.

[19]Narusaka, Y., Narusaka, M., Seki, M., et al., 2004. Crosstalk in the responses to abiotic and biotic stress in arabidopsis: analysis of gene expression in cytochrome P450 gene superfamily by cDNA microarray. Plant Mol. Biol., 55(3):327-342.

[20]Nelson, D.R., 2011. Progress in tracing the evolutionary paths of cytochrome P450. Biochem. Biophys. Acta, 1814(1):14-18.

[21]Ohkawa, H., Tsujii, H., Ohkawa, Y., 1999. The use of cytochrome P450 genes to introduce herbicide tolerance in crops: a review. Pestic. Sci., 55(9):867-874.

[22]Omokawa, H., Wu, J.R., Hatzios, K.K., 1996. Mechanism of safening action of dymuron and its two monomethyl analogues against bensulfuron-methyl injury to rice (Oryza sativa). Pestic. Biochem. Physiol., 55(1):54-63.

[23]Pan, G., Zhang, X.Y., Liu, K.D., et al., 2006. Map-based cloning of a novel rice cytochrome P450 gene CYP81A6 that confers resistance to two different classes of herbicides. Plant Mol. Biol., 61(6):933-943.

[24]Pang, S.S., Guddat, L.W., Duggleby, R.G., 2003. Molecular basis of sulfonylurea herbicide inhibition of acetohydroxyacid synthase. J. Biol. Chem., 278(9):7639-7644.

[25]Sterling, T.M., Balke, N.E., 1988. Use of soybean (Glycine max) and velvetleaf (Abutilon theopbrasti) suspension-cultured cells to study bentazon metabolism. Weed Sci., 36(5):558-568.

[26]Wang, Q.Z., Fu, H.W., Huang, J.Z., et al., 2012. Generation and characterization of bentazon susceptible mutants of commercial male sterile lines and evaluation of their utility in hybrid rice production. Field Crop. Res., 137: 12-18.

[27]Ward, E.R., Uknes, S.J., Williams, S.C., et al., 1991. Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell, 3(10):1085-1094.

[28]Yokosho, K., Yamaji, N., Ma, J.F., 2011. An Al-inducible MATE gene is involved in external detoxification of Al in rice. Plant J., 68(6):1061-1069.

[29]Yoo, S.D., Cho, Y.H., Sheen, J., 2007. Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat. Protoc., 2(7):1565-1572.

[30]Yuan, J.S., Tranel, P.J., Stewart, C.N.Jr., 2007. Non-target-site herbicide resistance: a family business. Trends Plant Sci., 12(1):6-13.

[31]Zhang, J., Xu, Y., Wu, X., et al., 2002. A bentazon and sulfonylurea sensitive mutant: breeding, genetics, and potential application in seed production of hybrid rice. Theor. Appl. Genet., 105(1):16-22.

[32]Zhang, L., Lu, Q., Chen, H.G., et al., 2006. Identification of a cytochrome P450 hydroxylase CYP81A6 as the candicate for the bentazon and sulfonylurea herbicide resistance gene, Bel, in rice. Mol. Breed., 19(1):59-68.

[33]Zhao, Y.D., 2010. Auxin biosynthesis and its role in plant development. Annu. Rev. Plant Biol., 61(1):49-64.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE