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1、作物学报ACTAAGRONOMICASINICA2016,42(4):513524http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2016.00513人工合成甘蓝型油菜及其亲本的甲基化变异模式分析*谢涛戎浩蒋金金孔月琴冉丽萍吴健王幼平扬州大学生物科学与技术学院,江苏扬州225009摘要:甘蓝型油菜作为多倍体起源和发生的历史较短,遗传背景较为狭窄,人工合成甘蓝型油菜可作为植物多倍化研究的优选模型,本文以人工
2、合成的甘蓝型油菜为材料,通过HPLC分析发现白菜型油菜和甘蓝的甲基化率分别为8.33%和15.88%,2个杂种株系的全基因组甲基化水平介于双亲之间,分别为10.29%和12.83%。MSAP分析发现杂种F1代及其亲本的甲基化水平存在明显差异(白菜型油菜<杂种F1<甘蓝),杂种F1代的甲基化变异(23.71%)中来自A、C基因组的变异分别占6.60%和10.16%。MSAP差异性条带的序列分析发现多倍化过程中与甲基化变化相关的基因参与了多种生物学过程,且差异甲基化基因在人工合成甘蓝型油菜及其亲本间的表达差异与甲基化修饰模式是一致的。本研究为了解甘蓝型油菜多倍化过程中发
3、生的表观变异奠定了基础。关键词:甘蓝型油菜;人工合成种;多倍化;甲基化AnalysisofDNAMethylationPatternsinResynthesizedBrassicanapusandDiploidParents*XIETao,RONGHao,JIANGJin-Jin,KONGYue-Qin,RANLi-Ping,WUJian,andWANGYou-PingCollegeofBioscienceandBiotechnology,YangzhouUniversity,Yangzhou225009,ChinaAbstract:Thegeneticbackgr
4、oundofBrassicanapus,oneofthemostimportantoilcropsinChina,isrelativelynarrowduetotheshorthistoryofitspolyploidorigin.ResynthesizedB.napusprovidesanoptimalmodelforresearchesonplantpolyploidization.Inthepresentstudy,wecomparedtheDNAmethylationlevelsinsynthesizedB.napus(F1generation)anddip
5、loidparentsusinghigh-performanceliquidchromatography(HPLC)andDNAmethylation-sensitiveamplificationpolymorphism(MSAP)analysis.HPLCanalysisindicatedmethylationratesof8.33%and15.88%inB.rapaandB.oleracea,respectively.Whilethemethylationrateoftwohybridswas10.29%and12.83%,whichwerein-between
6、oftheparents’values.MSAPanalysisprovedthedifferentmethylationlevelsinF1generationanddiploids,withthelowestandhighestmethylationlevelsidentifiedinB.rapaandB.ole-racea,respectively.VarianceoftheDNAmethylationinF1was23.71%,amongwhich6.60%and10.16%wereinheritedfromAandCgenome,respectively.
7、SequenceanalysisofMSAPpolymorphicfragmentsindicatedgenesinvolvedinmultiplemolecularfunctionswerechangedduringpolyploidization.Expressionanalysisofthesegenesagreedtothecorrespondingmethylationchanges.ThisstudyprovidespreliminarybasisforunderstandingepigeneticvariationsduringB.napuspol
