水稻P450基因Oscyp71Z2增强稻瘟病抗性的机制

doi:10.3864/j.issn.0578-1752.2014.13.001

Abstract

Abstract: 【Objective】The objective of this study is to analyze the role of cytochrome P450 gene Oscyp71Z2 in resistance to blast in rice, and expose the resistance mechanism mediated by Oscyp71Z2.【Method】Based on the transcriptome profile of the transgenic rice line NJH12 expressing hrf1 from Xoo, a resistance-related gene showing 114.6-fold increase of expression level was identified. The gene is the cytochrome P450 gene, named as Oscyp71Z2 (NCBI No.: Os07g0217600). Oscyp71Z2 coding sequence was cloned from rice cultivar Nipponbare transcript by PCR. To better understand the function of P450 gene Oscyp71Z2, binal transformation vectors pVec8::Oscyp71Z2 for Oscyp71Z2 overexpression were made, pVec8::Oscyp71Z2 plasmid was transformed into Agrobacterium tumefaciens strain EHA105 by freezing and thawing, and Oscyp71Z2-overexpressing rice was obtained by A. tumefaciens method. The selective marker hygromycin phosphotransferase gene hph was detected in T4 Oscyp71Z2-overexpressing rice by normal PCR, and the relative expression level of Oscyp71Z2 in transgenic rice was analyzed by qRT-PCR. Resistance phenotypes to Magnaporthe oryzae in positive Oscyp71Z2-overexpressing rice (T4, T5 and T6) were identified at booting stage. And phytoalexins accumulation of Oscyp71Z2-overexpressing rice (T4) was quantified.【Result】RT-PCR and qRT-PCR data showed that the expression level of cytochrome P450 gene Oscyp71Z2 in hrf1 transgenic rice with resistance to blast was higher than that in wild type rice, which was consistent with the Oscyp71Z2 data from transcriptome profile. Oscyp71Z2 gene cDNA is 1 554 bp, contains two cytochrome P450 domains by bioinformation analysis, which belongs to the CYP71Z subfamily. The full-length cDNA of Oscyp71Z2 was successfully ligated into binary vector pVec8, and transformed into rice Nipponbare genome mediated by A. tumefaciens. The rice growth periods, plant height and other traits in Oscyp71Z2-overexpressing rice were consistent with that in wild type rice. The transcripts of Oscyp71Z2 in Oscyp71Z2-overexpressing rice obviously increased, compared with that in wild type. The resistance degree to panicle blast in wild type was 3-4, showed susceptibility. However, the resistance degree to panicle blast in Oscyp71Z2-overexpressing rice was 1-2, showed resistance or middle resistance. The overexpression of Oscyp71Z2 in rice significantly enhanced the resistance to panicle blast. The accumulation levels of phytoalexin MA and MB in fresh leaves of Oscyp71Z2-overexpressing rice were 328 and 124 ng•g-1, respectively, which were higher than that of 85 and 42 ng•g-1 in wild type fresh leaves, respectively. Furthermore, the relative expression level of four key genes (OsCPS2, OsCPS4, OsKSL7 and OsKSL4) involve in phytoalexin MA and PB synthesis pathway in Oscyp71Z2-overexpressing rice was obviously increased.【Conclusion】The overexpression of cytochrome P450 gene Oscyp71Z2 in rice conferred resistance to panicle blast via activation of phytoalexins biosynthesis pathway.

Key words: rice , P450 gene Oscyp71Z2 , phytoalexins , disease resistance , blast

LI Wen-Qi-1, WANG Fang-Quan-1, WANG Jun-1, FAN Fang-Jun-1, ZHU Jin-Yan-1, YANG Jie-1, SHAO Min-2, ZHONG Wei-Gong-1. Study on Resistance Mechanism to Blast Mediated by P450 Gene Oscyp71Z2 in Rice[J].Scientia Agricultura Sinica, 2014, 47(13): 2485-2493.

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Study on Resistance Mechanism to Blast Mediated by P450 Gene Oscyp71Z2 in Rice

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