KY42表达可能可W增强水稻对稻癌病的抗性。
[0043] 2.遗传转化植株的共分离分析
[0044] 为了进一步验证上述推测，对在抗性增强的两个遗传转化植株值153RM9-6和 D153RM24-2)的T3家系进行基因表达量和抗性共分离分析。接种完稻癌病菌后，同时取样 抽取RNA，采用上述qRT-PCR分析方法检测植株中WRKY42基因的表达量。结果显示遗传转 化植株的W服Y42基因表达量与抗性共分离（图7);与对照牡丹江8相比所有抗性显著增强 的阳性转基因株系中的W服Y42基因的表达被抑制，而抗性与对照相比无显著变化的阴性 转基因植株中的W服Y42基因表达无显著变化。进一步证明遗传转化植株的抗性增强是因 为抑制WRKY42基因表达所至。送些研究结果也说明WRKY42基因的编码产物在水稻抗稻癌 病反应中发挥负调控因子的作用。抑制表达OsW服Y42基因，可增强水稻对稻癌病的抗性。 [004引参考文献
[0046] 曾亚，了新华，沈祥陵，李香花，王石平（2008)水稻抗病基因介导的抗白叶枯病反 应中蛋白质表达谱的比较分析。中国水稻科学22:230-238。
[0047]CaoY,DingX,CaiM,ZhaoJ,LinY,LiX,XuC,WangS(2007)Theexpression patternofaricediseaseresistancegeneXa3/Xa26isdifferentiallyregulated bythegeneticbackgroundsanddevelopmentalstagesthatinfluenceits function.Genetics177:523-533.
[0048]QienH,WangS,XingY,XuC,HayesPM,ZhangQ(2003)Comparativeanalysesof genomiclocationsandracespecificitiesoflociforquantitativeresistanceto Pyriculariagriseainriceandbarley.Proc.Natl.Acad.Sci.USA100:2544-2549.
[0049]ChuZ,OuyangY,ZhangJ,YangH,Wang8(2004)Genome-wideanalysisof defense-responsivegenesinbacterialblightresistanceofricemediatedbythe recessive民genexal3.Mol.Genet.Genomics271:111-120.
[0050]JonesJD,DanglJ"L(2006)Hieplantimmunesystem.Na化re444:323-329.
[0051]EulgemT,RushtonPJ,RobatzekS,Somssich1己（2000)化6W服Ys啤erfamilyof planttranscriptionfactors.TrendsPlantSci.5:199-206.
[0052]JiangJ,WangS口002)Identificationofa118libDNAfragmentcontaining thelocusofblastresistancegenePi-2 (t)inrice.Mol.Genet.Genomics 268:249-252.
[0053]KouY,WangS(2010)Broad-spectrumanddurability:understandingof quantitativediseaseresistance.Curr.Opin.PlantBiol. 13:181-185.
[0054]LiH,WangS(2013)Diseaseresistance.In:PlantGeneticsand Genomics:CropsandModelsVol. 5:GeneticsandGenomicsof民ice.EditedbyQifa Zhangand民odA.Wing.Springer,Heidelberg,p. 161-175.
[0055]LinYJ,ZhangQ(2005)Optimisingthetissuecultureconditionsforhigh efficiencytransformationofindicarice.PlantCell民ep. 23:540-547.
[0056]McGinnisK,ChandlerV，ConeK,KaepplerH,KaepplerS,KerschenA,Pikaard C,民ichardsE,SidorenkoL,SmithT,SpringerN，WulanT(2005)Transgene-inducedRNA interferenceasatoolforplantfunctionalgenomics.MethodsEnzymol. 392:1-24.
[0057]QiuD，XiaoJ，DingX，XiongM，CaiM，CaoY，LiX，XuC，Wang8(2007) OsWRKYlSmediatesricediseaseresistancebyregulatingdefense-related genesinsalicylate-andjasmonate-dependentsignaling.Mol.Plant-Microbe Interact. 20:492-499.
[0058]QiuD,XiaoJ,XieW,QiengH,LiX，Wang8(2009)E邱loringtranscriptional signalingmediatedbyOsWRKYlS,apotentialregulatorofmultiplephysiological processesinrice.BMCPlantBiol. 9:74.
[0059]SchenkPM,KazanK,WilsonI，AndersonJP,RichmondT,Somerville SC,MannersJM(2000)CoordinatedplantdefenseresponsesinArabidopsisrevealed bymicroarrayanalysis.Proc.Natl.Acad.Sci.USA97:11655-11660.
[0060]SmithNA,Sin組SP,WangMB,StoutjesdijkPA,GreenAG,WaterhousePM(2000) Geneexpression-Totalsilencingbyintron-splicedhairpin民NAs.Nature 407:319-320.
[0061]SunX，CaoY,YangZ,XuC,LiX，WangS,Slang Q(2004)Xa26,ageneconferring resistancetoXanthomonasoryzaepv.oryzaeinrice,encodingaL民民receptor kinase-likeprotein.PlantJ. 37:517-527.
[0062]ThommaBP,NurnbergerT，JoostenMH(2011)OfPAMPsandeffectors:the blurredPTI-ETIdichotomy.PlantCell23:4-15.
[0063]Wen化QiuZ,WangS(2003)Tlireetypesofdefense-responsivegenesare involvedinresistancetobacterialblightandfungalblastdiseasesinrice. Mol.Genet.Genomics269:331-339.
[0064]WilsonTA,Gough醒（1988)Hi組voltageE.colielectro-transformation withDNAfollowingligation.Nucl.Acids民es. 16:11820.
[006引 化anB,ShenX，LiX，XuC,WangS(2007)Mitogen-activatedproteinkinase 0sMPK6negativelyregulatesricediseaseresistancetobacterialpathogens.Planta226:953-960.
[0066]ZhangH,Wang8(2013)民iceversusXanthomonasoryzaepv.oryzae:aunique pathosystem.Curr.Opin.PlantBiol. 16:188-195.
[0067]ZhouB,PengKM,ChuZH,WangSP,ZhangQF(2002)化6defense-responsive genesshowingenhancedandrepressedexpressionafterpathogeninfectionin rice(OryzasativaL.).ScienceinChinaSeriesC-LifeSciences45:449-467.
【主权项】
1.核苷酸序列如序列表SEQIDNO:1中第1-946位碱基所示的WRKY42基因在增强水 稻对稻瘟病抗性中的应用。
【专利摘要】本发明涉及植物基因工程技术领域。具体涉水稻抗病相关基因WRKY42及在水稻改良中的应用。本发明包含水稻抗病相关基因WRKY42的功能验证。WRKY42基因表达量显著降低的遗传转化水稻对稻瘟病菌的抗性明显增强，证明WRKY42基因在水稻抗稻瘟病反应中是负调控因子，可以通过抑制WRKY42基因的表达改良水稻的抗病性。
【IPC分类】A01H5/00, C12N15/29, C12N15/82
【公开号】CN105368842
【申请号】CN201410438268
【发明人】王石平, 成洪涛
【申请人】华中农业大学
【公开日】2016年3月2日
【申请日】2014年8月29日