Transcriptome analysis.docx

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1、Tmnscrip1.onMu11i1.yh光拄花生EM中索的代妣径的输艮盘分析目的，I找到与，索合成相关的不因2attHua分析皿合成相网3明PKS基因在其中的作用InstructionWW第段：C6句)花生疮痂出凡客花生抡诟病(即加*“川MMWm(cBitaiKniir1.c1.Jcnkin)111940Bi1.iiucour1.泞次在阳关的巴西发现之后,日本根州宽山等I处9和阿根廷(GionhAH,1985)等国家也相维报道,并在各花生产区流行成灾近年来花?fc疮卷病在中国广东.广西、福建、山东和江字等主R花生产区相继发生他书.造成血枳OM产.危害产W.礁：段：花生疮痂桁曲同类桁吼式演相关

2、研究)花生疮琬病第是引欣桐再发生的物麻粉,盯性阶段为麴)堂腔牌,痂粪腔留本的生理特性Materia1.%a!nv(hn测量海表令最及f1.ifft的方法RNAPrCPn11ionftRNA的提双以及CDNA文库构建ToiaJRNAwasextractedfmfreshsc1.cttia1.ponderg喀TR1.zo11RC昨cn(试剂公司)accordingtoxnanutc1.urvr*sns1.rudxns.AMif1.ktcn1.atxun1.ofIu1.a1.1.!kIhanIOggwithOD2W,OD2SOva1.uesofI.Hto2.2Wused.Thewimp1.ewaxP

3、uriRcdandImaIcdwithFNaseinordertorr11vcPrO(CinandDNAContaminunts.Samp1.efort11incriponcana1.ysiswaspreparedasperI1.1.umiiumanufacturersinstructionsWhCrCmagticbeadswitho1.igodT)wereusedtoi%o1.a(eJxi1.yadeny1.a1.ednRNApc1.y(AHRNA)fromUieIuU1.RNA.Hugnien1.atiDnbufferCimxistingnfdiva1.entca1.in115was;nk

4、kxitxintcr1.inj:ERNAtoshandEBbut1.erforendrePantfiOnandUi1.infA.respective1.y.Puri1.1.cdCDMAfragmentswereconnectedwithWJUCrKinWadaptersa11eHforKRanp1.ifica1.n.AgI1.em2100BxQ1.UdyZeraidApp1.iedBkyaenH1.SICPomrPiURcaI-IutKPCRSy%11WefVUMfduqun1.iIicahmiMdqua1.1.ka1.!ofthesamp1.eIibran.forqua1.itycontro

5、1.PairedYndCDNA1.ib11r,w5constmctcdandMx1.I1.cIHrCdusing防序公司)Transvriptunwaa1.yxi转录fn测序以*39为材料，以谓控方素产生的主要环境闪索光照作为处理手区，分别以累暗处理和光照处理的碗姝作为林聚班的试发殂和对照机,DH1.aprocesi!数1处理方法ToavoidnegativeimpactonWbM:qucn1.bioinfma1.csana1.ysis,11wreadswerefi1.teredbytheremova1.ofraidswithaduptcrs.unknownnuc1.eo1.idcs1.arg

6、erthan10%.and1.owqua1.ityreadswheremorethanha1.fofthebasesqua1.itiesa1.essIhanfive(Q5).Basecompositionandqua1.itydis1.ributimWenrkIerniincdasqua1.itycn(1.I1.ieub1.awdc1.eantvadsvre1.1.enapfd1.IhegenometiiIgc11ct1.1.)cn%ax%cmb!y1.unnu1.a1.im功算生物侑息学分析Fora1.ternativesp1.icingana1.ysis,junctionsiteswere

7、detectedbyTopHutwithdefau1.tparamctc11i.JuiKiionsiteswhichtvidcinfrnaiiononIXHHk1.afieSandcmbiutonofdHcrencexonsinaIraiiMrnp1.Dr1.beunegeneWereutcdIUdistinguishtheIypeofaJIerm1.nv、p1.ingeentinc1.udingexondipping,intr11reniMn,a1.tcma1.ivc5sp1.icesite,andIkCgtiVC3sp1.icesite.Fornove1.transcriptspredic

8、tion,genemode1.sfoundinirue里CniCregions200basePaini(bp)uwayfromstreamordouiMreangenes)wcnwiththereferenceRCnnmCof1.rhinck5)andtheirfunctiona1.c1.assificationwasnnnoutcdbyGeneOnto1.ogy(GO).CI1.ftICrofOftho1.ogousGfOUPS.andKyotoEncyc1.opediaofGCnCandGCnOme9(KEGG1.PathWny.Conf1.rma1.kmfunicncexpression

9、usisrcH-ti11H*(unnthatiMrreversetranscriptionPCR使用实时荧光定址IjCR砒定韭因表达AIU1.1.yUofgcne%invo1.vedinmt，MKinrym*UM)4i5m对PKSAUMyHj分析光感应基因分析Kewit(hHictcrhticsc,riptknoftheproductionOCtoxinnnU)OrV光调控卜炳烧的点发的产生及产胞的特性描述RNAMH1.UenCin*aHM*Mt11wntRNA测序评估：测序的数1分析.RNAsamp1.ecIrac1.odfrnm1.rix0,Mi%ckm1.iumwaxXiibjcc1.c

10、dtohh-thr(Hghpu1.IHuminiisequencinginordertogainanoverviewofthefingusIranMJripcome.OVCr5GbofruwreadswereobtainedfromeachCDNA1.ibrary.TheSCqUenCi1.喀assessmentrevea1.edagoodsequencingqua1.ityadahgn11e1.tvsu1.1.Inkta1.52.933.332vadsWi1.h90bpinIeiig1.iiwereobtainedafterfi1.(enngIhe11wrcak.TheCICanreadwe

11、remappedtothegenomeAndgcno*cenccxof1.rtiinoccnxi$(Tab1.e1).Themappingresu1.tsprovidennovera1.1.asscssmcnof(besequencing.Acnandgenerespective1.yandabout6X.0and74.0%ofIberetptx1.ivemappedreudsIbepeftx1.mu1.chIyiUXXJ1.IiiiMiia1.ch).UIan0.5%IIK1.pPedivadsaremu1.hp1.iDna1.chwhi1.emajorityauuena1.ehwherthereadsWtnIrmappedtounique(on1.yone)positionsofthegenomeorgene.TherundomncssofmRNAfragmentationduringexperimenta1.settingwaseva1.uatedWi1.dtherandomdisibutinofreadsinreferencecncs.Apoorfadomncssoffragm