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.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2014.13.001

https://www.chinaagrisci.com/EN/Y2014/V47/I13/2485

References

[1]Talbot N J. On the trail of a cereal killer: exploring the biology of Magnaporthe grisea. Annual Review of Microbiology, 2003, 57: 177-202.

[2]王艳青. 近年来中国水稻病虫害发生及趋势分析. 中国农学通报, 2006, 22(2): 343-347.

Wang Y Q. Analysis on the occurrence and development of rice diseases and insects in China. Chinese Agricultural Science Bulletin, 2006, 22(2): 343-347. (in Chinese)

[3]Hasegawa M, Mitsuhara I, Seo S, Imai T, Koga J, Okada K, Yamane H, Ohashi Y. Phytoalexin accumulation in the interaction between rice and the blast fungus. Molecular Plant-Microbe Interactions, 2010, 8: 1000-1011.

[4]Koga J, Shimura M, Oshima K, Ogawa N, Yamauchi T, Ogasawara N. Phytocassanes A, B, C, and D, novel diterpene phytoalexins from rice, Oryza sativa L.. Tetrahedron, 1995, 51: 7907-7918.

[5]Koga J, Ogawa N, Yamauchi T, Kikuchi N, Ogasawara N, Shimura M. Functional moiety for the antifungal activity of phytocassane E, a diterpene phytoalexin from rice. Phytochemistry, 1997, 44: 249-253.

[6]Umemura K, Ogawa N, Yamauchi T, Iwata M, Shimura M, Koga J. Cerebroside elicitor found in diverse phytopathogens activate defense responses in rice plants. Plant Cell Physiology, 2000, 41: 676-683.

[7]Kenji U, Noriko O, Masaru S, Jinichiro K, Hideki U, Toshiaki K. Possible role of phytocassane, rice phytoalexin, in disease resistance of rice against the blast fungus Magnaporthe grisea. Bioscience Biotechnology and Biochemistry, 2003, 67: 899-902.

[8]Schuhegger R, Nafisi M, Mansourova M, Petersen B L, Olsen C E, Svatos A, Halkier B A, Glawischnig E. CYP71B15 (PAD3) catalyzes the final step in camalexin biosynthesis. Plant Physiology, 2006, 141(4): 1248-1254.

[9]Nafisi M, Goregaoker S, Botanga C J, Glawischnig E, Olsen C E, Halkier B A, Glazebrook J. Arabidopsis cytochrome P450 monooxygenase 71A13 catalyzes the conversion of indole-3- acetaldoxime in camalexin synthesis. The Plant Cell, 2007, 19: 2039-2052.

[10]Glawischnig E, Hansen B G, Olsen C E, Halkier B A. Camalexin is synthesized from indole-3-acetaldoxime, a key branching point between primary and secondary metabolism in Arabidopsis. Proceedings of the National Academy of Sciences of the USA, 2004, 101: 8245-8250.

[11]Li W Q, Shao M, Yang J, Zhong W G, Kazunori O, Hisakazu Y, Qian G L, Liu F Q. Oscyp71Z2 involves diterpenoid phytoalexin biosynthesis that contributes to bacterial blight resistance in rice. Plant Science, 2013, 207: 98-107.

[12]Shao M, Wang J S, Dean R A, Lin Y J, Gao X W, Hu S J. Expression of a harpin-encoding gene in rice confers durable nonspecific resistance to Magnaporthe grisea. Plant Biotechnology Journal, 2008, 6(1): 73-81.

[13]Li W Q, Shao M, Zhong W G, Yang J, Kazunori O, Hisakazu Y, Zhang L, Wang G, Wang D, Xiao S S, Chang S S, Qian G L, Liu F Q. Ectopic expression of hrf1 enhances bacterial resistance via regulation of diterpene phytoalexins, silicon and reactive oxygen species burst in rice. PLoS One, 2012, 7(9): e43914.

[14]邵敏, 吴智丹, 陈宝君, 落桑次仁, 李林. 转hrf1基因水稻对稻曲病抗性分析. 中国生物防治, 2008, 4: 335-338.

Shao M, Wu Z D, Chen B J, LuoSang C R, Li L. Resistance of hrf1 transgenic rice to Ustilaginoidea virens. Chinese Journal of Biological Control, 2008, 4: 335-338. (in Chinese)

[15]邵敏, 肖姗姗, 李林, 黄万春, 王金生. 转hrf1基因水稻对稻瘟病多小种非转化的稳定抗性. 中国水稻科学, 2008, 22(5): 459-464.

Shao M, Xiao S S, Li L, Huang W C, Wang J S. Expressing hrf1 gene in rice exhibits stable nonspecific resistance to Magnaporthe grisea. Chinese Journal of Rice Science, 2008, 22(5): 459-464. (in Chinese)

[16]Shimura K, Okada A, Okada K, Jikumaru Y, Ko K W, Toyomasu T, Sassa T, Hasegawa M, Kodama O, Shibuya N, Koga J, Nojiri H, Yamane H. Identification of a biosynthetic gene cluster in rice for momilactones. The Journal of Biological Chemistry, 2007, 282(11): 34013-34018.

[17]Swaminathan S, Morrone D, Wang Q, Fulton D B, Peters R J. CYP76M7 is an ent-cassadiene C11α-hydroxylase defining a second multifunctional diterpenoid biosynthetic gene cluster in rice. The Plant Cell, 2009, 21(10): 3315-3325.

[18]Cartwright D W, Langcake P, Pryce R J, Leworthy D P, Ride J P. Chemical activation of host defence mechanisms as a basis for crop protection. Nature, 1977, 267: 511-513.

[19]Kliebenstein D J, Rowe H C, Denby K J. Secondary metabolites influence Arabidopsis/Botrytis interactions: Variation in host production and pathogen sensitivity. The Plant Journal, 2005, 44: 25-36.

[20]Zhao Y, Hull A K, Gupta N R, Goss K A, Alonso J, Ecker J R, Normanly J, Chory J, Celenza J L. Trp-dependent auxin biosynthesis in Arabidopsis: Involvement of cytochrome P450s CYP79B2 and CYP79B3. Genes Development, 2002, 16(23): 3100-3112.

[21]Mikkelsen M D, Hansen C H, Wittstock U, Halkier B A. Cytochrome P450 CYP79B2 from Arabidopsis catalyzes the conversion of tryptophan to indole-3-acetal-doxime, a precursor of indole glucosinolates and indole-3-acetic acid. The Journal of Biological Chemistry, 2000, 275: 33712-33717.

[22]Glazebrook J, Ausubel F. Isolation of phytoalexin-deficient mutants of Arabidopsis thaliana and characterization of their interactions with bacterial pathogens. Proceedings of the National Academy of Sciences of the USA, 1994, 91: 8955-8959.

[23]Glazebrook J, Zook M, Mert F, Kagan I, Rogers E E, Crute I R, Holub E B, Hammerschmidt R, Ausubel F M. Phytoalexin-deficient mutants of Arabidopsis reveal that PAD4 encodes a regulatory factor and that four PAD genes contribute to downy mildew resistance. Genetics, 1997, 146: 381-392.

[24]Kim J A, Cho K, Singh R, Jung Y H, Jeong S H, Kim S H, Lee J E, Cho Y S, Agrawal G K, Rakwal R, Tamogami S, Kersten B, Jeon J S, An G, Jwa N S. Rice OsACDR1 (Oryza sativa Accelerated Cell Death and Resistance 1) is a potential positive regulator of fungal disease resistance. Molecular Cells, 2009, 8: 431-439.

[25]Wang Q, Hillwig M L, Peters R J. CYP99A3: Functional identification of a diterpene oxidase from the momilactone biosynthetic gene cluster in rice. The Plant Journal, 2011, 65(1): 87-95.

[26]Nelson D R, Schuler M A, Paquette S M, Werck-Reichhart D, Bak S. Comparative genomics of rice and Arabidopsis. Analysis of 727 cytochrome P450 genes and pseudogenes from a monocot and a dicot. Plant Physiology, 2004, 135(2): 756-772.

Related Articles 15

[1]	XIAO DeShun, XU ChunMei, WANG DanYing, ZHANG XiuFu, CHEN Song, CHU Guang, LIU YuanHui. Effects of Rhizosphere Oxygen Environment on Phosphorus Uptake of Rice Seedlings and Its Physiological Mechanisms in Hydroponic Condition [J]. Scientia Agricultura Sinica, 2023, 56(2): 236-248.
[2]	ZHANG XiaoLi, TAO Wei, GAO GuoQing, CHEN Lei, GUO Hui, ZHANG Hua, TANG MaoYan, LIANG TianFeng. Effects of Direct Seeding Cultivation Method on Growth Stage, Lodging Resistance and Yield Benefit of Double-Cropping Early Rice [J]. Scientia Agricultura Sinica, 2023, 56(2): 249-263.
[3]	ZHANG Wei,YAN LingLing,FU ZhiQiang,XU Ying,GUO HuiJuan,ZHOU MengYao,LONG Pan. Effects of Sowing Date on Yield of Double Cropping Rice and Utilization Efficiency of Light and Heat Energy in Hunan Province [J]. Scientia Agricultura Sinica, 2023, 56(1): 31-45.
[4]	FENG XiangQian,YIN Min,WANG MengJia,MA HengYu,CHU Guang,LIU YuanHui,XU ChunMei,ZHANG XiuFu,ZHANG YunBo,WANG DanYing,CHEN Song. Effects of Meteorological Factors on Quality of Late Japonica Rice During Late Season Grain Filling Stage Under ‘Early Indica and Late Japonica’ Cultivation Pattern in Southern China [J]. Scientia Agricultura Sinica, 2023, 56(1): 46-63.
[5]	SANG ShiFei,CAO MengYu,WANG YaNan,WANG JunYi,SUN XiaoHan,ZHANG WenLing,JI ShengDong. Research Progress of Nitrogen Efficiency Related Genes in Rice [J]. Scientia Agricultura Sinica, 2022, 55(8): 1479-1491.
[6]	GUI RunFei,WANG ZaiMan,PAN ShengGang,ZHANG MingHua,TANG XiangRu,MO ZhaoWen. Effects of Nitrogen-Reducing Side Deep Application of Liquid Fertilizer at Tillering Stage on Yield and Nitrogen Utilization of Fragrant Rice [J]. Scientia Agricultura Sinica, 2022, 55(8): 1529-1545.
[7]	LIAO Ping,MENG Yi,WENG WenAn,HUANG Shan,ZENG YongJun,ZHANG HongCheng. Effects of Hybrid Rice on Grain Yield and Nitrogen Use Efficiency: A Meta-Analysis [J]. Scientia Agricultura Sinica, 2022, 55(8): 1546-1556.
[8]	HAN XiaoTong,YANG BaoJun,LI SuXuan,LIAO FuBing,LIU ShuHua,TANG Jian,YAO Qing. Intelligent Forecasting Method of Rice Sheath Blight Based on Images [J]. Scientia Agricultura Sinica, 2022, 55(8): 1557-1567.
[9]	GAO JiaRui,FANG ShengZhi,ZHANG YuLing,AN Jing,YU Na,ZOU HongTao. Characteristics of Organic Nitrogen Mineralization in Paddy Soil with Different Reclamation Years in Black Soil of Northeast China [J]. Scientia Agricultura Sinica, 2022, 55(8): 1579-1588.
[10]	ZHU DaWei,ZHANG LinPing,CHEN MingXue,FANG ChangYun,YU YongHong,ZHENG XiaoLong,SHAO YaFang. Characteristics of High-Quality Rice Varieties and Taste Sensory Evaluation Values in China [J]. Scientia Agricultura Sinica, 2022, 55(7): 1271-1283.
[11]	GUO ZeXi,SUN DaYun,QU JunJie,PAN FengYing,LIU LuLu,YIN Ling. The Role of Chalcone Synthase Gene in Grape Resistance to Gray Mold and Downy Mildew [J]. Scientia Agricultura Sinica, 2022, 55(6): 1139-1148.
[12]	WANG Kai,ZHANG HaiLiang,DONG YiXin,CHEN ShaoKan,GUO Gang,LIU Lin,WANG YaChun. Definition and Genetic Parameters Estimation for Health Traits by Using on-Farm Management Data in Dairy Cattle [J]. Scientia Agricultura Sinica, 2022, 55(6): 1227-1240.
[13]	ZHAO Ling, ZHANG Yong, WEI XiaoDong, LIANG WenHua, ZHAO ChunFang, ZHOU LiHui, YAO Shu, WANG CaiLin, ZHANG YaDong. Mapping of QTLs for Chlorophyll Content in Flag Leaves of Rice on High-Density Bin Map [J]. Scientia Agricultura Sinica, 2022, 55(5): 825-836.
[14]	JIANG JingJing,ZHOU TianYang,WEI ChenHua,WU JiaNing,ZHANG Hao,LIU LiJun,WANG ZhiQin,GU JunFei,YANG JianChang. Effects of Crop Management Practices on Grain Quality of Superior and Inferior Spikelets of Super Rice [J]. Scientia Agricultura Sinica, 2022, 55(5): 874-889.
[15]	ZHANG YaLing, GAO Qing, ZHAO Yuhan, LIU Rui, FU Zhongju, LI Xue, SUN Yujia, JIN XueHui. Evaluation of Rice Blast Resistance and Genetic Structure Analysis of Rice Germplasm in Heilongjiang Province [J]. Scientia Agricultura Sinica, 2022, 55(4): 625-640.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

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

RichHTML

PDF

Cited