水稻细菌性条斑病菌中分泌蛋白质组表达谱的研究
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摘要
水稻细菌性条斑病菌(Xamthomonas.oryzae pv.oryzicola,Xoc)是水稻的主要病原菌之一,其引发的水稻细菌性条斑病可造成水稻严重减产。本研究以Xoc GX01作为实验菌株,通过细菌培养、分泌蛋白提取、纯化获得总分泌蛋白,将样品酶解后通过LC-MS/MS技术鉴定了分泌蛋白质表达谱,共鉴定蛋白质661个,同时利用生物信息学技术对鉴定蛋白进行基因本体(gene ontology,GO)生物学分类,对生物功能、生物进程以及细胞组件进行了说明。为阐明植物病原菌的作用机理以及植物-病原细菌互作提供了理论基础。
Xamthomonas.oryzae pv.oryzicola(Xoc) is one of the main pathogens,of rice which can cause severe reduction of rice yield.In this study,Xoc GX01 was used as the experimental strain.We got the total secreted proteins by bacterial culture,secreted protein extraction and purification.Then,the secreted protein expression spectrum was identified by LC-MS/MS after enzymatic hydrolysis of the sample,and 661 proteins were identified.Meanwhile,bioinformatics technology was used to carry on the gene ontology classification for identified proteins,and to explain the biological function,biological process and cellular component.This result might provide the theoretical basis for explaining the acting mechanism of the plant pathogenic bacteria and plant-pathogenic bacteria interactions.
Xamthomonas.oryzae pv.oryzicola(Xoc) is one of the main pathogens,of rice which can cause severe reduction of rice yield.In this study,Xoc GX01 was used as the experimental strain.We got the total secreted proteins by bacterial culture,secreted protein extraction and purification.Then,the secreted protein expression spectrum was identified by LC-MS/MS after enzymatic hydrolysis of the sample,and 661 proteins were identified.Meanwhile,bioinformatics technology was used to carry on the gene ontology classification for identified proteins,and to explain the biological function,biological process and cellular component.This result might provide the theoretical basis for explaining the acting mechanism of the plant pathogenic bacteria and plant-pathogenic bacteria interactions.
引文
Andrade A.E.,Silva L.P.,Pereira J.L.,Noronha E.F.,Reis F.B.,Bloch C.,dos Santos M.F.,Domont G.B.,Franco O.L.,and Mehta A.,2008,In vivo proteome analysis of Xanthomonas campestris pv.campestris in the interaction with the host plant Brassica oleracea,FEMS Microbiol Let,281(2):167-174
Koevar N.,Grazio S.F.,and Komel R.,2014,Two-dimensional gel electrophoresis of gastric tissue in an alkaline p H range,Proteomics,14(2-3):311-321
Kim S.G.,Wang Y.,Lee K.H.,Park Z.Y.,Park J.,Wu J.,Kwon S.J.,Lee Y.H.,Agrawal G.K.,Rakwal R.,Kim S.T.,and Kang K.Y.,2013,In-depth insight into in vivo apoplastic secretome of rice-Magnaporthe oryzae interaction,Proteomics,78:58-71
Krause C.,Richter S.,Knoll C.,and Jrgens G.,2013,Plant secretome-from cellular process to biological activity,Biochim Biophys Acta,1834(11):2429-2441
Li B.Q.,Wang W.H.,and Zong Y.Y.,2012,Exploring pathogenic mechanisms of Botrytis cinerea secretome under different ambient p H based on comparative proteomic analysis,J.Proteome Res,11(8):4249-4260
Qian G.L.,Zhou Y.J.,Zhao Y.C.,Song Z.W.,Wang S.Y.,Fan J.Q.,Hu B.S.,Venturi V.,and Liu F.Q.,2013,Proteomic analysis reveals novel extracellular virulence-associated proteins and functions regulated by the diffusible signal factor(DSF)in Xanthomonas oryzae pv.oryzicola,J.Proteome Res,12:3327-3341
Shah P.,Gutierrez-Sanchez G.,and Orlando R.,2009,A proteomic study of pectin-degrading enzymes secreted by Botrytis cinerea grown in liquid culture,Proteomics,9(11):3126-3135
Stephanowitz H.,Lange S.,Lang D.,Freund C.,and Krause E.,2012,Improved two-dimensional reversed phasereversed phase LC-MS/MS approach for identification of peptide-protein interactions,J.Proteome Res.,11:1175-1183
Wang J.Z.,Shi L.M.,He X.P.,Lu L.D.,Li X.P.,and Chen B.S.,2016,Comparative secretome analysis reveals perturbation of host secretion pathways by a hypovirus,Sci.Rep.,6:34308
Wang Y.,Wu J.,Park Z.Y.,Kim S.G.,Rakwal R.,Agrawal G.K.,Kim S.T.,and Kang K.Y.,2011,Comparative secretome investigation of Magnaporthe oryzae proteins responsive to nitrogen starvation,Journal of Proteome Research,10(7):3136-3148
Xiao Y.S.,Wei X.X.,Gao H.P.,Cen Z.L.,Huang P.,Wu Y.,Wang R.,and He Y.Q.,2011,Characterization of a Xanthomonas oryzae pv.oryzicola strain and the establishment of its genetic manipulation system,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),30(33):1211-1217(肖永胜,韦雪雪,郜惠苹,牛祥娜,岑贞陆,黄萍,吴玥,汪嵘,何勇强,2011,一株水稻细菌性条斑病菌的鉴定与遗传操作系统的建立,基因组学与应用生物学,30(33):1211-1217)
Zhao Y.C.,Qian G.L.,Yin F.Q.,Fan J.Q.,Zhai Z.W.,Liu C.H.,Hu B.S.,and Liu F.Q.,2011,Proteomic analysis of the regulatory function of DSF-dependent quorum sensing in Xanthomonas oryzae pv.Oryzicola,Microbial Pathogenesis 50:48-55
Zhu P.C.,Qin H.P.,Fu S.,Cen W.L.,Zou H.F.,He Y.Q.,and Jiang W.,2015,The Research of Xamthomonas.oryzae pv.oryzicola protein expression spectrum based on LC-MS-MStechnology,Jiyinzuxue Yu Yingyong Shegnwuxue(Genomics and Applied Biology),34(6):1239-1244(朱平川,覃宏平,付珊,岑卫健,邹海凡,何勇强,姜伟,2015,基于LC-MS-MS技术建立水稻细菌性条斑病菌的蛋白质表达谱的研究,基因组学与应用生物学,34(6):1239-1244)
Zou H.S.,Yuan L.,Guo W.,Li Y.R.,Che Y.Z.,Zou L.F.,and Chen G.Y.,2011,Construction of a Tn5-tagged mutant library of Xanthomonas oryzae pv.oryzicola as an invaluable resource for functional genomics,62(3):908-916
Koevar N.,Grazio S.F.,and Komel R.,2014,Two-dimensional gel electrophoresis of gastric tissue in an alkaline p H range,Proteomics,14(2-3):311-321
Kim S.G.,Wang Y.,Lee K.H.,Park Z.Y.,Park J.,Wu J.,Kwon S.J.,Lee Y.H.,Agrawal G.K.,Rakwal R.,Kim S.T.,and Kang K.Y.,2013,In-depth insight into in vivo apoplastic secretome of rice-Magnaporthe oryzae interaction,Proteomics,78:58-71
Krause C.,Richter S.,Knoll C.,and Jrgens G.,2013,Plant secretome-from cellular process to biological activity,Biochim Biophys Acta,1834(11):2429-2441
Li B.Q.,Wang W.H.,and Zong Y.Y.,2012,Exploring pathogenic mechanisms of Botrytis cinerea secretome under different ambient p H based on comparative proteomic analysis,J.Proteome Res,11(8):4249-4260
Qian G.L.,Zhou Y.J.,Zhao Y.C.,Song Z.W.,Wang S.Y.,Fan J.Q.,Hu B.S.,Venturi V.,and Liu F.Q.,2013,Proteomic analysis reveals novel extracellular virulence-associated proteins and functions regulated by the diffusible signal factor(DSF)in Xanthomonas oryzae pv.oryzicola,J.Proteome Res,12:3327-3341
Shah P.,Gutierrez-Sanchez G.,and Orlando R.,2009,A proteomic study of pectin-degrading enzymes secreted by Botrytis cinerea grown in liquid culture,Proteomics,9(11):3126-3135
Stephanowitz H.,Lange S.,Lang D.,Freund C.,and Krause E.,2012,Improved two-dimensional reversed phasereversed phase LC-MS/MS approach for identification of peptide-protein interactions,J.Proteome Res.,11:1175-1183
Wang J.Z.,Shi L.M.,He X.P.,Lu L.D.,Li X.P.,and Chen B.S.,2016,Comparative secretome analysis reveals perturbation of host secretion pathways by a hypovirus,Sci.Rep.,6:34308
Wang Y.,Wu J.,Park Z.Y.,Kim S.G.,Rakwal R.,Agrawal G.K.,Kim S.T.,and Kang K.Y.,2011,Comparative secretome investigation of Magnaporthe oryzae proteins responsive to nitrogen starvation,Journal of Proteome Research,10(7):3136-3148
Xiao Y.S.,Wei X.X.,Gao H.P.,Cen Z.L.,Huang P.,Wu Y.,Wang R.,and He Y.Q.,2011,Characterization of a Xanthomonas oryzae pv.oryzicola strain and the establishment of its genetic manipulation system,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),30(33):1211-1217(肖永胜,韦雪雪,郜惠苹,牛祥娜,岑贞陆,黄萍,吴玥,汪嵘,何勇强,2011,一株水稻细菌性条斑病菌的鉴定与遗传操作系统的建立,基因组学与应用生物学,30(33):1211-1217)
Zhao Y.C.,Qian G.L.,Yin F.Q.,Fan J.Q.,Zhai Z.W.,Liu C.H.,Hu B.S.,and Liu F.Q.,2011,Proteomic analysis of the regulatory function of DSF-dependent quorum sensing in Xanthomonas oryzae pv.Oryzicola,Microbial Pathogenesis 50:48-55
Zhu P.C.,Qin H.P.,Fu S.,Cen W.L.,Zou H.F.,He Y.Q.,and Jiang W.,2015,The Research of Xamthomonas.oryzae pv.oryzicola protein expression spectrum based on LC-MS-MStechnology,Jiyinzuxue Yu Yingyong Shegnwuxue(Genomics and Applied Biology),34(6):1239-1244(朱平川,覃宏平,付珊,岑卫健,邹海凡,何勇强,姜伟,2015,基于LC-MS-MS技术建立水稻细菌性条斑病菌的蛋白质表达谱的研究,基因组学与应用生物学,34(6):1239-1244)
Zou H.S.,Yuan L.,Guo W.,Li Y.R.,Che Y.Z.,Zou L.F.,and Chen G.Y.,2011,Construction of a Tn5-tagged mutant library of Xanthomonas oryzae pv.oryzicola as an invaluable resource for functional genomics,62(3):908-916