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.ehwherth
12、ereadsWtnIrmappedtounique(on1.yone)positionsofthegenomeorgene.TherundomncssofmRNAfragmentationduringexperimenta1.settingwaseva1.uatedWi1.dtherandomdisibutinofreadsinreferencecncs.Apoorfadomncssoffragmcn3on1.eadstogenera1.ionofreadsfromspecificregion%ofUieorigina1.Uunscripbandai1.edingsubsequentana1.
13、ysis.InIhUMudy.IbCdh(rib,readsin1.hinoccrtis.Atota1.of88.2(9.475of10.742)“gee%werecovedby(1.ein;IpPCdread%f11theInsiscnpfDinekt(asetw1.wrvH3.0%ofthegenwXhoWedpcrtcctcoverageof90to100%.AboUt97.0%ofgenescoverageishigherthan50%,Atou1.ofH.64I91.2%)transcrip(o11cgcwIDoddSwerereassigneduponrcfmcnofgenestr
14、ucturesadakcmaiivcSP1.iCmgAS)ana1.ysis格从曲捱取的RNA样品进行高通所IIIumHU测序,以狭得人照转录组的提述.从你个CDNA文蚱抉得超这5Gb的班如读(,测序评估般示了乩Qj的测序Mi1.it和出准结果.在过泄Svtti.总共获得长度为90即的52.933332个读纭CTearread映射到该式倒留第因投和播区库对衣I).-MttWI户上mT我。1.rf*IM1.5#与测绘结果提供源序的总体评估.总共65.4%和46.1%的C1.earread铤绢分别陵射到典曲|州知密*41&些闪讥和压因，大的68.0和74.。的各个映射的读取得到完美匹配(没药不匹配
15、).少于0.5%的映射读取是多位双也配，而大多数足堆或配，犬中读取故映射到她囚班或塔囚的堆Y仪一个）.在参考基闪中读取的前机分布泮估丈验设置期何mRNA片段的Mi机性。碎片的1.机性较筮守致从Ki始转录物的特定区域产生读数并彬响就后的分析,在这项研允中，读i的分布是均匀的,具右良好的碎片随机性（图1A）.因此我明了良好的序列数据.图IB显示了此类真1中韭因1?施率（读故援盍的城因百分比）的分布.来自转来:加数据集的映射读数躅鱼/Ja共882%”0.742个）中的9.475个）IM,其中83Q%的基因显示完全IS菠率为90至100%.妁97Q%的M因理能率修于50%.在压因结构和选择性剪接（AS
16、分析的优化后,共转录了8.64）个（91.2）转录仪取因模型。BOtnri*BCMMGenome-basedfunctiona1.an*,JiW*1.M(rtrtW景用不同方式对转乘组进行功饯注释1.KEGC注Wi“Of)fKUrtMWXf4AMBOyourCJtro*MS)ftoytmpwnv*MM5w3rvJCMkSAUWV)*JAUOOU*JJOUOQXix*su*vmqMoq0uJcuruoMu09n9pcwRUDmu4RMOMowai9bCXwwyeMHPuiUCvMUOMUOKMteM、CVtfM0UMMMJV1fUfXmw9UTModuJM3IVWwWputJdbuv4muQm
17、9hWMoqHw(3UM9VMmV0OUfM)Qn2PWPtOi：JMHOQTHWC*U0jwUm0kjmj(HwpurmMUVNUY3RO93ZRgIWVm.0f,-pe0nvwpEDoaXBuOs!nbut11dOOanptbonC1.uiarComponentMocu1.arFunctionBGIOg1.Ca1.ProcsCOUoeOJOqEnN1.Vga*sodatcdENondarymetnboIi1.egincs与毒力相关的次f1.t代置若B92.Viruk*nceassciatvdsigna1.ingpathways毒力相关的他!号K5.（eteaxMciatcdithdc（oM
18、catim与作用相关的茶因4.Phto%e11xitivcgeneIPmtein）与光相关的基因（光W白）5.Gencsre1.atedtosporeproduction。产施相关的机囚,SCDreductase.Bn1.SODDNAhdIKnDNArpMrproteinrd50PMi、.narby1.x1.r*t*eu*cy1.trMa11a*pcc4rvare1.iedIoPETSptncnCaIIM”itMycyv1.ewewoeucpo*VhMdccdPbO4Ut皿betaIeOMMTiMHoadouMuncxAmmWCuIf1.ocvUti40pt(4eaFXO1.1.Suiuiar
19、to*ubMu*!ikesetmeprXcMtMNiribosomaipeJIDnhyhnBKftrasetype11eo51CoAhyvUMinihrIotafuindi1.f)odipoimivn1.haKettyIbwwfevweNApMk1.TnamaM，&h)dru.NADdepcudoaiCPnnetWjddt)9*fn!yproteinNAW*)ttdcdey9Mucro9etmc12cUm11aooM*fcrie8atno*oxc*ounoMecytfhaecUFaci1.itatorSupcHHniI)(oo*terMFSaf1.atoxinff1.tttIM1.nPMwFK
20、iIrttforSiRgiibMFStvwMpoderMFS11atidnranpcofxinfIMOtrinctcfetIraiMpoi1.et!m1.yfvctatnma9ne4mbampo11crAUUMkia41UMtdtMocUv1OMkxbMKbva1.MpoHI1.g0*tdxmtram1.MeMIbUUtt三!0rodMetec1.1.cyc1.ec1.po*ptutaseHicyiMehyxy1.ase17beUhygMrcHd(MdroJmUerUtedomaMia三oprotein1JK(MKyYCyiYVnfffrc*in)rttaMBdndx(NADPH)Bochc*
21、ia1.ATPMduwe9pk三cninnt-Mifvot*inkyQyacy1ddy8ofcg4miCTMtmatmI.mi1.OdMM1.uro1.oxdmep63uitdc沪UOOtoxdfcductuc.f1.ocbcAiINeCurXrC6ZmCfrn*etdooMinoa1.reetnC6tracr10afctocC6trantcofdxTPMDNAvttdRNADohTOtfivePKSsSPCCirka1.beXPreSSCdat(heHahtCOditiOn卷过以上坛IS信息的分析,确定H的施囚在虻环境下表达差异即显寿。谷项代谢通路Phyh)HcnCtiCandmainanjyxc%ffn1.PCIykrtideM,nthi5c%(PKS).明确PKS然因的种类三三:c三三M三三一三t三wI1.三1.三三三a二二二三三三(nePnfnaryNna1.ofsduiyingtheCnnmCofafun新1.PatbOgen1、toidentifyMXandarymet;IboIHrIhaiarcSCrVCdasViRIICnefactorssuchashostspecifictoxins.nonhastspecifictoxin(NHST)udme1.anin.JR.可行性分析图片.数据分析).璋素含量，地子St
