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二斑叶螨不同发育时期基因组DNA甲基化水平的研究.docx

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二斑叶螨不同发育时期基因组DNA甲基化水平的研究 Title:GenomeDNAMethylationLevelinDifferentDevelopmentStagesofTwo-SpottedSpiderMite Introduction: Two-spottedspidermite(Tetranychusurticae)isoneofthemostharmfulpestsofvariouscrops,causingsignificantyieldlossesglobally.Inrecentyears,thestudyofepigeneticmodificationhasbecomeanessentialresearchareainthefieldofpestcontrol.DNAmethylationisanessentialmolecularmechanismofepigeneticregulation,whichcanaffectgeneexpressionandplayacrucialroleinregulatingbiologicalprocesses.However,littleisknownaboutthegenomeDNAmethylationlevelintwo-spottedspidermites.Here,wefocusedonthegenomeDNAmethylationlevelindifferentdevelopmentstagesoftwo-spottedspidermites. Materialsandmethods: Inthisstudy,wecollectedT.urticaefromalaboratorycolonymaintainedonkidneybeanplants.Threedevelopmentalstages,includingeggs,larvae,andadults,wereharvested.DNAwasisolatedusingtheCTABmethodandsequencedonthePacBioRSIIplatform.DNAmethylationlevelswereestimatedusingthemethylpypackage.ThesignificanceofmethylationdifferencesamongdifferentstageswasanalyzedusingtheT-test. Results: Weobtained6,576,409,276sequencingreadsforT.urticae,andthegenomesizewas90.51Mb.Amongthem,0.33%,0.36%,and0.31%ofthegenomeweremethylatedineggs,larvae,andadults,respectively.Themethylationlevelsofnon-codingregions(includingintergenicregions,introns,anduntranslatedregions)weresignificantlyhigherthanthoseofcodingregions(includingexonsandCDS).Andthehighestmethylationlevelwasdetectedinintrons.Inaddition,wefoundsomemethylationsitesshowedsignificantdifferencesamongdifferentdevelopmentalstages.Moreover,themethylationlevelsofsomegenes,especiallythegenesrelatedtostressresponseandimmunity,variedindifferentdevelopmentstages. Discussion: TheresultsshowedthatT.urticaehadalowDNAmethylationlevel,whichwasconsistentwithsomepreviousstudiesonmites.Themethylationlevelsofnon-codingregionswerehigherthanthoseincodingregions,whichmightbeinfluencedbythefunctionalconstraintofcodingregions.Notably,thehighmethylationlevelofintronsmightplayaroleingeneregulat