一个抗病性增强的水稻类病变突变体的蛋白质组学研究

韩雪颖1,2 ,杨勇2 ,余初浪2 ,张文浩3 ,叶胜海2 ,陈斌2 ,程晨2 ,程晔2 ,严成其2 ,陈剑平1,2,*

doi:10.3969/j.issn.1001-7216.2014.06. 001

中国水稻科学 >

2014 , Vol. 28 >Issue 6: 559 - 569

DOI: https://doi.org/10.3969/j.issn.1001-7216.2014.06. 001

研究报告

一个抗病性增强的水稻类病变突变体的蛋白质组学研究

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¹南京农业大学植物保护学院，南京 210095；²浙江省植物有害生物防控重点实验室——省部共建国家重点实验室培育基地/农业部植保生物技术重点实验室/浙江省植物病毒学重点实验室/浙江省农业科学院病毒学与生物技术研究所，杭州 310021；³中国计量学院生命科学学院，杭州310018；

收稿日期: 2014-04-16

修回日期: 2014-06-27

网络出版日期: 2014-11-10

基金资助

国家973计划资助项目(2012CB722504)；国家自然科学基金青年项目(31101208, 31201482)；浙江省自然科学基金重点项目(Z14C140001)。

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A Proteomic Study on a DiseaseResistanceEnhanced Rice Lesion Mimic Mutant

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¹College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; ²State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Key Laboratory of Biotechnology in Plant Protection, MOA, Zhejiang Provincial Key Laboratory of Plant Virology, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Science, Hangzhou 310021, China; ³College of Life Science, China Jiliang University, Hangzhou 310018, China;

Received date: 2014-04-16

Revised date: 2014-06-27

Online published: 2014-11-10

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摘要

水稻类病变突变体chl1具有抗病性增强的类病变表型，对水稻白叶枯病和稻瘟病都具有很强的抗性。利用蛋白质组学技术分析chl1与其野生型之间的差异表达蛋白，探讨chl1类病变表型的形成和抗病反应的分子机制。利用荧光双向差异凝胶电泳(twodimensional fluorescence difference gel electrophoresis, 2DDIGE)技术和质谱分析，chl1中共鉴定到70个差异表达的蛋白点，包括46个上调蛋白点和24个下调蛋白点。这些蛋白点参与不同的生物过程，包括防御相关、光合作用、氧化还原、氨基酸/蛋白质代谢、分子伴侣、碳水化合物代谢。对这些差异表达蛋白进行生物信息分析，推测它们所在的复杂调控网络可能参与chl1叶片细胞程序性死亡(programmed cell death, PCD)及其抗病性的调控。

关键词： 水稻; 类病变突变体; 细胞程序性死亡; 抗病性; 荧光双向差异凝胶电脉

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韩雪颖1,2 ,杨勇2 ,余初浪2 ,张文浩3 ,叶胜海2 ,陈斌2 ,程晨2 ,程晔2 ,严成其2 ,陈剑平1,2,* . 一个抗病性增强的水稻类病变突变体的蛋白质组学研究[J]. 中国水稻科学, 2014 , 28(6) : 559 -569 . DOI: 10.3969/j.issn.1001-7216.2014.06. 001

Abstract

A lesionmimic mutant in rice (Oryza sativa L.), chloroplasticH2O2induced lesion 1 (chl1), has enhanced resistance to rice blast and bacterial blight. To understand the molecular mechanisms underlying this phenotype and its resistance responses, a proteomicsbased approach was used to identify differentiallyexpressed proteins between chl1 and its wild type. Using twodimensional fluorescence difference gel electrophoresis technology and mass spectrometry, 70 protein spots were successfully identified, of which 46 were upregulated and 24 were downregulated in the mutant. These differentiallyexpressed proteins are involved in diverse biological processes including disease resistance, photosynthesis, oxidationreduction reaction, amino acid/protein metabolism, chaperoning and carbohydrate metabolism. The complex regulatory network in which these proteins are involved may play an important role in regulating the programmed cell death and the resistance reaction in chl1.

Key words： rice; lesionmimic mutant; programmed cell death; disease resistance; 2DDIGE

参考文献

\[1\]Morel J B, Dangl J L. The hypersensitive response and the induction of cell death in plants. Cell Dea Dif, 1997, 4(8): 671683.

\[2\]Hoeberichts F A, Woltering E J. Multiple mediators of plant programmed cell death: Interplay of conserved cell death mechanisms and plantspecific regulators. BioEssays, 2003, 25(1): 4757.

\[3\]Johal G S, Hulbert S H, Briggs S P. Disease lesion mimics of maize: A model for cell death in plants. BioEssays, 1995, 17(8): 685692.

\[4\]Wolter M, Hollricher K, Salamini F, et al. The mlo resistance alleles to powdery mildew infection in barley trigger a developmentally controlled defence mimic phenotype. Mol Gen Gene, 1993, 239(12): 122128.

\[5\]Dietrich R A, Delaney T P, Uknes S J, et al. Arabidopsis mutants simulating disease resistance response. Cell, 1994, 77(4): 565577.

\[6\]Mizobuchi R, Hirabayashi H, Kaji R, et al. Isolation and characterization of rice lesionmimic mutants with enhanced resistance to rice blast and bacterial blight. Plant Sci, 2002, 163(2): 345353.

\[7\]陈析丰, 金杨, 马伯军. 水稻类病变突变体及抗病性的研究进展. 植物病理学报, 2011, 41(1): 19.

\[8\]Lorrain S, Vailleau F, Balagué C, et al.Lesion mimic mutants: Keys for deciphering cell death and defense pathways in plants? Trends Plant Sci, 2003, 8(6): 263271.

\[9\]Wu C, Bordeos A, Madamba M R, et al. Rice lesion mimic mutants with enhanced resistance to diseases. Mol Genet Genom, 2008, 279(6): 605619.

\[10\]Weeks M E, Sinclair J, Jacob R J, et al. Stressinduced changes in the Schizosaccharomyces pombe proteome using twodimensional difference gel electrophoresis, mass spectrometry and a novel integrated robotics platform. Proteomics, 2005, 5(6): 16691685.

\[11\]Alfonso P,Núez A, MadozGurpide J, et al. Proteomic expression analysis of colorectal cancer by twodimensional differential gel electrophoresis. Proteomics, 2005, 5(10): 26022611.

\[12\]Chen X, Fu S, Zhang P, et al.Proteomic analysis of a diseaseresistanceenhanced lesion mimic mutant spotted leaf 5 in rice. Rice, 2013, 6(1): 110.

\[13\]Deng Z, Zhang X, Tang W, et al.A proteomics study of brassinosteroid response in Arabidopsis. Mol Cell Proteom, 2007, 6(12): 20582071.

\[14\]Kang S G, Matin M N, Bae H, et al. Proteome analysis and characterization of phenotypes of lesion mimic mutant spotted leaf 6 in rice. Proteomics, 2007, 7(14): 24472458.

\[15\]Tonge R, Shaw J, Middleton B, et al.Validation and development of fluorescence twodimensional differential gel electrophoresis proteomics technology. Proteomics, 2001, 1(3): 377396.

\[16\]Takahashi A, Kawasaki T, Wong H L, et al. Hyperphosphorylation of a mitochondrial protein, prohibitin, is induced by calyculin A in a rice lesionmimic mutant cdr1. Plant Physiol, 2003, 132(4): 18611869.

\[17\]成晓越. 水稻类病变新基因 chl1 (chloroplasticH2O2induced lesion 1) 的鉴定与克隆.金华：浙江师范大学， 2013.

\[18\]胡明, 李晓宇, 马平, 等. 抗真菌蛋白研究进展. 生物技术通报, 2004, 3: 1317.

\[19\]Van Loon L, Rep M, Pieterse C. Significance of inducible defenserelated proteins in infected plants.Ann Rev Phytopathol, 2006, 44: 135162.

\[20\]Jung Y H, Rakwal R, Agrawal G K, et al. Differential expression of defense/stressrelated marker proteins in leaves of a unique rice blast lesion mimic mutant (blm). J Proteome res, 2006, 5(10): 25862598.

\[21\]Tsunezuka H, Fujiwara M, Kawasaki T, et al. Proteome analysis of programmed cell death and defense signaling using the rice lesion mimic mutant cdr2. Mol Plantmicro Intera, 2005, 18(1): 5259.

\[22\]Kim S T, Kim S G, Kang Y H, et al. Proteomics analysis of rice lesion mimic mutant (spl1) reveals tightly localized probenazoleinduced protein (PBZ1) in cells undergoing programmed cell death. J Proteome Res, 2008, 7(4): 17501760.

\[23\]Bai S, Dong C, Li B,et al. A PR4 gene identified from Malus domestica is involved in the defense responses against Botryosphaeria dothidea. Plant Physiol Biochem, 2013, 62: 2332.

\[24\]Van Loon L, Pierpoint W, Boller T, et al. Recommendations for naming plant pathogenesisrelated proteins. Plant Mol Biol Rep, 1994, 12(3): 245264.

\[25\]Datta K, Velazhahan R, Oliva N, et al. Overexpression of the cloned rice thaumatinlike protein (PR5) gene in transgenic rice plants enhances environmental friendly resistance to Rhizoctonia solani causing sheath blight disease. Theor Appl Genet, 1999, 98(67): 11381145.

\[26\]Wang Y, Wu J, Lee D Y,et al. Comparative secretome analysis of differentiallyinduced proteins in rice lesion mimic mutant spotted leaf 11 (spl11). Plant Om, 2012, 5(6): 567575.

\[27\]Kim S G, Kim S T, Wang Y,et al. The RNase activity of rice probenazoleinduced protein1 (PBZ1) plays a key role in cell death in plants. Mol Cells, 2011, 31(1): 2531.

\[28\]李红丽, 刘迪秋, 何华, 等. 类萌发素蛋白在植物防卫反应中的作用. 植物生理学报, 2013, 49(4): 331336.

\[29\]Dixon D P, Lapthorn A, Edwards R. Plant glutathione transferases. Genom Biol, 2002, 3(3): 3004.13004.10.

\[30\]裴冬丽. 谷胱甘肽还原酶在植物防御中的研究进展. 中国农学通报, 2012, 28(18): 185188.

\[31\]王玉, 孔凡英, 尹波, 等. 过表达单脱氢抗坏血酸还原酶基因提高番茄抗UVB胁迫能力. 植物生理学报, 2014, 50(1): 95104.

\[32\]Yoon H S, Lee H, Lee I, et al. Molecular cloning of the monodehydroascorbate reductase gene from Brassica campestris and analysis of its mRNA level in response to oxidative stress. Biochim Biophysi Acta, 2004, 1658(3): 181186.

\[33\]梅杨, 李海蓝, 谢晋, 等. 核酮糖1,5二磷酸羧化酶/加氧酶 (Rubisco). 植物生理学通讯, 2007, 43(2): 363.

\[34\]Pearrubia L, Moreno J. Increased susceptibility of ribulose1,5bisphosphate carboxylase/oxygenase to proteolytic degradation caused by oxidative treatments. Archiv biochem biophy, 1990, 281(2): 319323.

\[35\]Sedigheh H G, Mortazavian M, Norouzian D,et al. Oxidative stress and leaf senescence. BMC Res notes, 2011, 4(1): 477485.

\[36\]Zhao C, Wang J, Cao M,et al. Proteomic changes in rice leaves during development of fieldgrown rice plants. Proteomics, 2005, 5(4): 961972.

\[37\]王强, 温晓刚, 张其德. 光合作用光抑制的研究进展. 植物学通报, 2004, 20(5): 539548.

\[38\]Heide H, Kalisz H M, Follmann H. The oxygen evolving enhancer protein 1 (OEE) of photosystem II in green algae exhibits thioredoxin activity.J Plant physiol, 2004, 161(2): 139149.

\[39\]Gururani M A, Upadhyaya C P, Strasser R J, et al. Evaluation of abiotic stress tolerance in transgenic potato plants with reduced expression of PSII manganese stabilizing protein. Plant Sci, 2013, 198: 716.

\[40\]Liu Y, Ren D, Pike S, et al. Chloroplastgenerated reactive oxygen species are involved in hypersensitive responselike cell death mediated by a mitogenactivated protein kinase cascade. Plant J, 2007, 51(6): 941954.

\[41\]Gao L, Yan X, Li X,et al. Proteome analysis of wheat leaf under salt stress by twodimensional difference gel electrophoresis (2DDIGE). Phytochemistry, 2011, 72(10): 11801191.

\[42\]Ndimba B K, Chivasa S, Simon W J,et al. Identification of Arabidopsis salt and osmotic stress responsive proteins using twodimensional difference gel electrophoresis and mass spectrometry. Proteomics, 2005, 5(16): 41854196.

\[43\]Parker R, Flowers T J, Moore A L, et al. An accurate and reproducible method for proteome profiling of the effects of salt stress in the rice leaf lamina. J Exp Bot, 2006, 57(5): 11091118.

\[44\]杨洪强, 高华君. 植物精氨酸及其代谢产物的生理功能. 植物生理与分子生物学学报, 2007, 33(1): 18.

\[45\]Gonzalez M E, Marco F, Minguet E G,et al. Perturbation of spermine synthase gene expression and transcript profiling provide new insights on the role of the tetraamine spermine in Arabidopsis defense against Pseudomonas viridiflava. Plant Physiol, 2011, 156(4): 22662277.

\[46\]Zhang H, Liu W, Wan L, et al. Functional analyses of ethylene response factor JERF3 with the aim of improving tolerance to drought and osmotic stress in transgenic rice. Transg Res, 2010, 19(5): 809818.

\[47\]Ondzighi C A, Christopher D A, Cho E J,et al. Arabidopsis protein disulfide isomerase5 inhibits cysteine proteases during trafficking to vacuoles before programmed cell death of the endothelium in developing seeds. Plant Cell, 2008, 20(8): 22052220.

\[48\]陈珍, 江琼, 朱诚. 植物中的蛋白质二硫键异构酶及其类蛋白. 植物生理学报, 2013, 49(8): 715721.

\[49\]吴厚雄, 肖辉海, 李必湖. 植物热激蛋白的研究进展. 生物技术通报, 2003(4): 69.

\[50\]秦佳, 杨金莹, 伊淑莹, 等. 热激蛋白对细胞凋亡的调节作用. 生命科学, 2007, 19(2): 159163.

\[51\]Qi Y, Wang H, Zou Y,et al. Overexpression of mitochondrial heat shock protein 70 suppresses programmed cell death in rice. FEBS Let, 2011, 585(1): 231239.

\[52\]Ito M, Yamamoto Y, Kim C S, et al. Heat shock protein 70 is required for tabtoxinineβlactaminduced cell death in Nicotiana benthamiana. J Plant Physiol, 2014, 171(2): 173178.

\[53\]Kanzaki H, Saitoh H, Ito A,et al. Cytosolic HSP90 and HSP70 are essential components of INF1mediated hypersensitive response and nonhost resistance to Pseudomonas cichorii in Nicotiana benthamiana. Mol Plant Pathol, 2003, 4(5): 383391.

\[54\]Lu W, Tang X, Huo Y,et al. Identification and characterization of fructose 1,6bisphosphate aldolase genes in Arabidopsis reveal a gene family with diverse responses to abiotic stresses. Gene, 2012, 503(1): 6574.

\[55\]Chen M, Mishra S, Heckathorn S A,et al. Proteomic analysis of Arabidopsis thaliana leaves in response to acute boron deficiency and toxicity reveals effects on photosynthesis, carbohydrate metabolism, and protein synthesis. J Plant physiol, 2014, 171(3): 235242.

\[56\]Kirsch M, De Groot H. NAD(P)H, a directly operating antioxidant. FASEB J, 2001, 15(9): 15691574.

\[57\]Nakamura H.Thioredoxin and its related molecules. Ant red sign, 2005, 7(5/6): 823828.

\[58\]Leterrier M, Barroso J B, Valderrama R,et al. NADPdependent isocitrate dehydrogenase from Arabidopsis roots contributes in the mechanism of defence against the nitrooxidative stress induced by salinity. Sci World J, 2012, 2012: 694740.

\[59\]Martinoia E, Rentsch D. Malate compartmentationresponses to a complex metabolism.Ann Rev Plant Biol, 1994, 45(1): 447467.

\[60\]Wang Q F, Zhao Y, Yi Q, et al. Overexpression of malate dehydrogenase in transgenic tobacco leaves: Enhanced malate synthesis and augmented Alresistance. Acta Physiol Plant, 2010, 32(6): 12091220.

\[61\]魏爱丽, 王志敏, 翟志席, 等. 土壤干旱对小麦旗叶和穗器官C4光合酶活性的影响. 中国农业科学, 2003, 36(5): 508512.

\[62\]Fu Z, Zhang Z, Liu Z, et al. The effects of abiotic stresses on the NADPdependent malic enzyme in the leaves of the hexaploid wheat. Biol Plant, 2011, 55(1): 196200.

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