芸薹根肿菌侵染早期甘蓝型油菜转录组分析
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摘要
根肿病是油菜最为严重的病害之一。为探索根肿病菌和油菜互作中的抗性分子机制,采用RNA-seq技术对根肿病早期侵染中抗病品系ZHE-226(R)和感病油菜10159(S)进行转录组分析。与感病材料S相比,在0、12、48和72h 4个接种时间点,抗性材料R中共有809个基因上调表达,1082个基因下调表达。模式识别受体、几丁质酶、R基因、WRKY、水杨酸和茉莉酸等抗性相关(PRR)基因在抗病材料R和感病材料S中诱导表达并表现出不同的表达模式。抗病材料R中PRR相关基因主要表现为下调表达,R基因上调表达,水杨酸表现为前期下调后期上调表达,茉莉酸、几丁质酶和WRKY因子在R和S中均诱导表达。结果表明PRR介导的PTI(病原相关分子模式触发的免疫反应)在感病材料S中显著诱导,在抗病材料R中作用不明显; R基因介导的ETI(效应蛋白触发的免疫反应)和水杨酸介导的抗病信号途径对根肿病抗性起重要作用。
Clubroot is one of the most serious disease in oilseed rape.To explore the resistant molecular mechanism of the interaction between Plasmodiophora brassicae and Brassica napus during early infection,gene differential expression analysis of the clubroot-resistant rapeseed line ZHE-226(R) and clubroot-susceptible line10159(S) were performed by RNA-seq.A total of 809 and 1082 genes were commonly up-and down-regulated at 0,12,48,72 h post-inoculation(hpi) time points in R line.Pattern recognition receptors(PRRs),chitinase,R gene,WRKY transcription factor,salicylic acid(SA) and jasmonic acid(JA) were involved in the early response to clubroot infection in lines of R and S,but showed different expression patterns.Compared to S line,most of the PRRs genes were down-regulated,R genes were up-regulated and SA genes were down-regulated in earlier stage but up-regulated at 72 hpi in R line.Chitinase,WRKY and JA related genes were both induced in R and S lines.These results showed that PTI-mediated by PRRs was significantly induced in S line,but played a small role in R line.ETI-mediated by R gene and resistance signal pathway-mediated by SA played great role in clubroot resistance.This research provided a better understanding of the resistance genes of B.napus in response to P.brassicae,and assistance for clubroot-resistant breeding.
Clubroot is one of the most serious disease in oilseed rape.To explore the resistant molecular mechanism of the interaction between Plasmodiophora brassicae and Brassica napus during early infection,gene differential expression analysis of the clubroot-resistant rapeseed line ZHE-226(R) and clubroot-susceptible line10159(S) were performed by RNA-seq.A total of 809 and 1082 genes were commonly up-and down-regulated at 0,12,48,72 h post-inoculation(hpi) time points in R line.Pattern recognition receptors(PRRs),chitinase,R gene,WRKY transcription factor,salicylic acid(SA) and jasmonic acid(JA) were involved in the early response to clubroot infection in lines of R and S,but showed different expression patterns.Compared to S line,most of the PRRs genes were down-regulated,R genes were up-regulated and SA genes were down-regulated in earlier stage but up-regulated at 72 hpi in R line.Chitinase,WRKY and JA related genes were both induced in R and S lines.These results showed that PTI-mediated by PRRs was significantly induced in S line,but played a small role in R line.ETI-mediated by R gene and resistance signal pathway-mediated by SA played great role in clubroot resistance.This research provided a better understanding of the resistance genes of B.napus in response to P.brassicae,and assistance for clubroot-resistant breeding.
引文
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[9] Feng J,Hwang S F,Strelkov S E. Studies into primary and secondary infection processes by Plasmodiophora brassicae on canola[J]. Plant Pathol,2013,62(1):177-183.
[10] Deora A,Gossen B D,McDonald M R. Infection and development of Plasmodiophora brassicae in resistant and susceptible canola cultivars[J]. Can J Plant Pathol,2012,34(2):239-247.
[11] Kageyama K,Asano T. Life cycle of Plasmodiophora brassicae[J]. J Plant Growth Regul,2009,28(3):203-211.
[12] Devos S,Vissenberg K,Verbelen J P,et al. Infection of Chinese cabbage by Plasmodiophora brassicae leads to a stimulation of plant growth:impacts on cell wall metabolism and hormone balance[J]. New Phytol,2004,166(1):241-250.
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[14] Sun L Q,Zhu L F,Xu L,et al. Cotton cytochrome P450 CYP82D regulates systemic cell death by modulating the octadecanoid pathway[J]. Nat Commun,2014,5:5372.
[15]陈英,谭碧玥,黄敏仁.植物天然免疫系统研究进展[J].南京林业大学学报(自然科学版),2012,36(1):129-136.
[16] Zhao Y,Bi K,Gao Z X,et al. Transcriptome analysis of Arabidopsis thaliana in response to Plasmodiophora brassicae during early infection[J]. Front Microbiol,2017,8:673.
[17] Chen J J,Pang W X,Chen B,et al. Transcriptome analysis of Brassica rapa near-isogenic lines carrying clubroot-resistant and–susceptible alleles in response to Plasmodiophora brassicae during early infection[J].Front Plant Sci,2016,6:1183.
[18] Siemens J,Keller I,Sarx J,et al. Transcriptome analysis of Arabidopsis clubroots indicate a key role for cytokinins in disease development[J]. Mol Plant-Microbe Interactions,2006,19(5):480-494.
[19] Schuller A,Kehr J,Ludwig-Müller J. Laser microdissection coupled to transcriptional profiling of Arabidopsis roots inoculated by Plasmodiophora brassicae indicates a role for brassinosteroids in clubroot formation[J]. Plant Cell Physiol,2014,55(2):392-411.
[20] Agarwal A,Kaul V,Faggian R,et al. Analysis of global host gene expression during the primary phase of the Arabidopsis thaliana-Plasmodiophora brassicae interaction[J]. Funct Plant Biol,2011,38(6):462.
[21] Jubault M,Hamon C,Gravot A,et al. Differential regulation of root arginine catabolism and polyamine metabolism in clubroot-susceptible and partially resistant Arabidopsis genotypes[J]. Plant Physiol,2008,146(4):2008-2019.
[22] Chu M G,Song T,Falk K C,et al. Fine mapping of Rcr1 and analyses of its effect on transcriptome patterns during infection by Plasmodiophora brassicae[J]. BMC Genom,2014,15(1):1166.
[23]战宗祥,江莹芬,朱紫媛,等.与位点Pb Ba8. 1紧密连锁分子标记的开发及甘蓝型油菜根肿病抗性育种[J].中国油料作物学报,2015,37(6):766-771.
[24] Chen J J,Jing J,Zhan Z X,et al. Identification of novel QTLs for isolate-specific partial resistance to Plasmodiophora brassicae in Brassica rapa[J]. PLo S One,2013,8(12):e85307.
[25]杨佩文,李家瑞,杨勤忠,等.十字花科蔬菜根肿病菌休眠孢子的分离与检测[J].云南农业大学学报,2002,17(3):301-302,306.
[26] Love M I,Huber W,Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J]. Genome Biol,2014,15(12):550.
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[28] The Gene Ontology Consortium. Expansion of the gene ontology knowledgebase and resources[J]. Nucleic Acids Res,2017,45:331-338.
[29] Kanehisa M. The KEGG resource for deciphering the genome[J]. Nucleic Acids Res,2004,32(90001):277-280.
[30] Chen C,Xia R,Chen H,et al. TBtools,a Toolkit for biologists integrating various HTS-data handling tools with a user-friendly interface[J]. bioRxiv,2018:289660.
[31] Cao T S,Srivastava S,Rahman M H,et al. Proteome-level changes in the roots of Brassica napus as a result of Plasmodiophora brassicae infection[J]. Plant Sci,2008,174(1):97-115.
[32] Hatakeyama K,Suwabe K,Tomita R N,et al. Identification and characterization of Crr1a,a gene for resistance to clubroot disease(Plasmodiophora brassicae Woronin)in Brassica rapa L[J]. PLo S One,2013,8(1):e54745.
[33] Zhang T,Zhao Z,Zhang C Y,et al. Fine genetic and physical mapping of the CRb gene conferring resistance to clubroot disease in Brassica rapa[J]. Mol Breeding,2014,34(3):1173-1183.
[34] Monaghan J,Zipfel C. Plant pattern recognition receptor complexes at the plasma membrane[J]. Curr Opin Plant Biol,2012,15(4):349-357.
[35] Li C Y,Deng G M,Yang J,et al. Transcriptome profiling of resistant and susceptible Cavendish banana roots following inoculation with Fusarium oxysporum f. sp.cubense tropical race 4[J]. BMC Genom,2012,13(1):374.
[36]糜艳霞,任慧,张常,等.几丁质酶的研究进展[J].生命科学研究,2015,19(5):437-443.
[37]田义,康国栋,张彩霞,等.几丁质触发植物免疫的研究现状与展望[J].中国农业科学,2013,46(15):3115-3124.
[38]范伟,李雪姣,关明俐,等.水稻几丁质酶基因的转录与表达特征[J].作物学报,2014,40(4):571-580.
[39]张志忠,吴菁华,吕柳新,等.植物几丁质酶及其应用研究进展[J].福建农业大学学报,2005,34(4):494-499.
[40]张志忠,吴菁华,吕柳新,等.植物几丁质酶及其应用研究进展[J].福建农林大学学报(自然科学版),2005,34(4):494-499.
[41]贾翠玲,侯和胜.植物WRKY转录因子的结构特点及其在植物防卫反应中的作用[J].天津农业科学,2010,16(2):21-26.
[42]陈思雀,翁群清,曹红瑞,等. WRKY转录因子在生物和非生物胁迫中的功能和调控机理的研究进展[J].农业生物技术学报,2017,25(4):668-682.
[43] Deslandes L,Olivier J,Theulieres F,et al. Resistance to Ralstonia solanacearum in Arabidopsis thaliana is conferred by the recessive RRS1-R gene,a member of a novel family of resistance genes[J]. Proc Natl Acad Sci,2002,99(4):2404-2409.
[44] Knoth C,Ringler J,Dangl J L,et al. Arabidopsis WRKY70 is required for FullRPP4-mediated disease resistance and basal defense against Hyaloperonospora parasitica[J]. Mol Plant-Microbe Interactions,2007,20(2):120-128.
[45] Kim K C,Fan B,Chen Z. Pathogen-induced Arabidopsis WRKY7 is a transcriptional repressor and enhances plant susceptibility to Pseudomonas syringae[J].Plant Physiol,2006,142(3):1180-1192.
[46] Eulgem T,Somssich I E. Networks of WRKY transcription factors in defense signaling[J]. Curr Opin Plant Biol,2007,10(4):366-371.
[47]房卫平,谢德意,李志芳,等. NBS-LRR类抗病蛋白介导的植物抗病应答分子机制[J].分子植物育种,2015,13(2):469-474.
[48] Zhang X L,Liu Y M,Fang Z Y,et al. Comparative transcriptome analysis between broccoli(Brassica oleracea var. italica)and wild cabbage(Brassica macrocarpa Guss.)in response to Plasmodiophora brassicae during different infection stages[J]. Front Plant Sci,2016,7:1929.
[49] Irani S,Trost B,Waldner M,et al. Transcriptome analysis of response to Plasmodiophora brassicae infection in the Arabidopsis shoot and root[J]. BMC Genom,2018,19:23.
[50] Glazebrook J. Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens[J]. Annu Rev Phytopathol,2005,43(1):205-227.
[2] Piao Z Y,Ramchiary N,Lim Y P. Genetics of clubroot resistance in Brassica species[J]. J Plant Growth Regul,2009,28(3):252-264.
[3] Schwelm A,Fogelqvist J,Knaust A,et al. The Plasmodiophora brassicae genome reveals insights in its life cycle and ancestry of chitin synthases[J]. Sci Rep,2015,5:11153.
[4] Burki F,Kudryavtsev A,Matz M V,et al. Evolution of Rhizaria:new insights from phylogenomic analysis of uncultivated protists[J]. BMC Evol Biol,2010,10(1):377.
[5]王靖,黄云,胡晓玲,等.油菜根肿病症状、病原形态及产量损失研究[J].中国油料作物学报,2008,30(1):112-115.
[6]季海雯,任莉,陈坤荣,等.油菜根肿病病原主要生理小种和品种抗病性鉴定[J].中国油料作物学报,2013,35(3):301-306.
[7] Kowata-Dresch L S,May-De Mio L L. Clubroot management of highly infested soils[J]. Crop Prot,2012,35:47-52.
[8] Donald C,Porter I. Integrated control of clubroot[J]. J Plant Growth Regul,2009,28(3):289-303.
[9] Feng J,Hwang S F,Strelkov S E. Studies into primary and secondary infection processes by Plasmodiophora brassicae on canola[J]. Plant Pathol,2013,62(1):177-183.
[10] Deora A,Gossen B D,McDonald M R. Infection and development of Plasmodiophora brassicae in resistant and susceptible canola cultivars[J]. Can J Plant Pathol,2012,34(2):239-247.
[11] Kageyama K,Asano T. Life cycle of Plasmodiophora brassicae[J]. J Plant Growth Regul,2009,28(3):203-211.
[12] Devos S,Vissenberg K,Verbelen J P,et al. Infection of Chinese cabbage by Plasmodiophora brassicae leads to a stimulation of plant growth:impacts on cell wall metabolism and hormone balance[J]. New Phytol,2004,166(1):241-250.
[13] Jones J D G,Dangl J L. The plant immune system[J].Nature,2006,444(7117):323-329.
[14] Sun L Q,Zhu L F,Xu L,et al. Cotton cytochrome P450 CYP82D regulates systemic cell death by modulating the octadecanoid pathway[J]. Nat Commun,2014,5:5372.
[15]陈英,谭碧玥,黄敏仁.植物天然免疫系统研究进展[J].南京林业大学学报(自然科学版),2012,36(1):129-136.
[16] Zhao Y,Bi K,Gao Z X,et al. Transcriptome analysis of Arabidopsis thaliana in response to Plasmodiophora brassicae during early infection[J]. Front Microbiol,2017,8:673.
[17] Chen J J,Pang W X,Chen B,et al. Transcriptome analysis of Brassica rapa near-isogenic lines carrying clubroot-resistant and–susceptible alleles in response to Plasmodiophora brassicae during early infection[J].Front Plant Sci,2016,6:1183.
[18] Siemens J,Keller I,Sarx J,et al. Transcriptome analysis of Arabidopsis clubroots indicate a key role for cytokinins in disease development[J]. Mol Plant-Microbe Interactions,2006,19(5):480-494.
[19] Schuller A,Kehr J,Ludwig-Müller J. Laser microdissection coupled to transcriptional profiling of Arabidopsis roots inoculated by Plasmodiophora brassicae indicates a role for brassinosteroids in clubroot formation[J]. Plant Cell Physiol,2014,55(2):392-411.
[20] Agarwal A,Kaul V,Faggian R,et al. Analysis of global host gene expression during the primary phase of the Arabidopsis thaliana-Plasmodiophora brassicae interaction[J]. Funct Plant Biol,2011,38(6):462.
[21] Jubault M,Hamon C,Gravot A,et al. Differential regulation of root arginine catabolism and polyamine metabolism in clubroot-susceptible and partially resistant Arabidopsis genotypes[J]. Plant Physiol,2008,146(4):2008-2019.
[22] Chu M G,Song T,Falk K C,et al. Fine mapping of Rcr1 and analyses of its effect on transcriptome patterns during infection by Plasmodiophora brassicae[J]. BMC Genom,2014,15(1):1166.
[23]战宗祥,江莹芬,朱紫媛,等.与位点Pb Ba8. 1紧密连锁分子标记的开发及甘蓝型油菜根肿病抗性育种[J].中国油料作物学报,2015,37(6):766-771.
[24] Chen J J,Jing J,Zhan Z X,et al. Identification of novel QTLs for isolate-specific partial resistance to Plasmodiophora brassicae in Brassica rapa[J]. PLo S One,2013,8(12):e85307.
[25]杨佩文,李家瑞,杨勤忠,等.十字花科蔬菜根肿病菌休眠孢子的分离与检测[J].云南农业大学学报,2002,17(3):301-302,306.
[26] Love M I,Huber W,Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J]. Genome Biol,2014,15(12):550.
[27] Ashburner M,Ball C A,Blake J A,et al. Gene Ontology:tool for the unification of biology[J]. Nat Genet,2000,25(1):25-29.
[28] The Gene Ontology Consortium. Expansion of the gene ontology knowledgebase and resources[J]. Nucleic Acids Res,2017,45:331-338.
[29] Kanehisa M. The KEGG resource for deciphering the genome[J]. Nucleic Acids Res,2004,32(90001):277-280.
[30] Chen C,Xia R,Chen H,et al. TBtools,a Toolkit for biologists integrating various HTS-data handling tools with a user-friendly interface[J]. bioRxiv,2018:289660.
[31] Cao T S,Srivastava S,Rahman M H,et al. Proteome-level changes in the roots of Brassica napus as a result of Plasmodiophora brassicae infection[J]. Plant Sci,2008,174(1):97-115.
[32] Hatakeyama K,Suwabe K,Tomita R N,et al. Identification and characterization of Crr1a,a gene for resistance to clubroot disease(Plasmodiophora brassicae Woronin)in Brassica rapa L[J]. PLo S One,2013,8(1):e54745.
[33] Zhang T,Zhao Z,Zhang C Y,et al. Fine genetic and physical mapping of the CRb gene conferring resistance to clubroot disease in Brassica rapa[J]. Mol Breeding,2014,34(3):1173-1183.
[34] Monaghan J,Zipfel C. Plant pattern recognition receptor complexes at the plasma membrane[J]. Curr Opin Plant Biol,2012,15(4):349-357.
[35] Li C Y,Deng G M,Yang J,et al. Transcriptome profiling of resistant and susceptible Cavendish banana roots following inoculation with Fusarium oxysporum f. sp.cubense tropical race 4[J]. BMC Genom,2012,13(1):374.
[36]糜艳霞,任慧,张常,等.几丁质酶的研究进展[J].生命科学研究,2015,19(5):437-443.
[37]田义,康国栋,张彩霞,等.几丁质触发植物免疫的研究现状与展望[J].中国农业科学,2013,46(15):3115-3124.
[38]范伟,李雪姣,关明俐,等.水稻几丁质酶基因的转录与表达特征[J].作物学报,2014,40(4):571-580.
[39]张志忠,吴菁华,吕柳新,等.植物几丁质酶及其应用研究进展[J].福建农业大学学报,2005,34(4):494-499.
[40]张志忠,吴菁华,吕柳新,等.植物几丁质酶及其应用研究进展[J].福建农林大学学报(自然科学版),2005,34(4):494-499.
[41]贾翠玲,侯和胜.植物WRKY转录因子的结构特点及其在植物防卫反应中的作用[J].天津农业科学,2010,16(2):21-26.
[42]陈思雀,翁群清,曹红瑞,等. WRKY转录因子在生物和非生物胁迫中的功能和调控机理的研究进展[J].农业生物技术学报,2017,25(4):668-682.
[43] Deslandes L,Olivier J,Theulieres F,et al. Resistance to Ralstonia solanacearum in Arabidopsis thaliana is conferred by the recessive RRS1-R gene,a member of a novel family of resistance genes[J]. Proc Natl Acad Sci,2002,99(4):2404-2409.
[44] Knoth C,Ringler J,Dangl J L,et al. Arabidopsis WRKY70 is required for FullRPP4-mediated disease resistance and basal defense against Hyaloperonospora parasitica[J]. Mol Plant-Microbe Interactions,2007,20(2):120-128.
[45] Kim K C,Fan B,Chen Z. Pathogen-induced Arabidopsis WRKY7 is a transcriptional repressor and enhances plant susceptibility to Pseudomonas syringae[J].Plant Physiol,2006,142(3):1180-1192.
[46] Eulgem T,Somssich I E. Networks of WRKY transcription factors in defense signaling[J]. Curr Opin Plant Biol,2007,10(4):366-371.
[47]房卫平,谢德意,李志芳,等. NBS-LRR类抗病蛋白介导的植物抗病应答分子机制[J].分子植物育种,2015,13(2):469-474.
[48] Zhang X L,Liu Y M,Fang Z Y,et al. Comparative transcriptome analysis between broccoli(Brassica oleracea var. italica)and wild cabbage(Brassica macrocarpa Guss.)in response to Plasmodiophora brassicae during different infection stages[J]. Front Plant Sci,2016,7:1929.
[49] Irani S,Trost B,Waldner M,et al. Transcriptome analysis of response to Plasmodiophora brassicae infection in the Arabidopsis shoot and root[J]. BMC Genom,2018,19:23.
[50] Glazebrook J. Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens[J]. Annu Rev Phytopathol,2005,43(1):205-227.