5个湖泊河川沙塘鳢粒体细胞色素b基因的遗传变异

J．LakeSci．2015274693-699DOI10．18307/2015．04182015byJournalofLakeSciences5Odontobutispotamophilab*1121111＊＊121001723130015DNAbPCＲ、1141bp．cytbA+T55．8%G+C44．2%806115870．969±0．0120．20081±0．00742．0．1370．424．Fst0．531．cytbUPGMA5．DNAbGeneticvariationanalysisofOdontobutispotamophilafromfivelakesbasedonMitochon-drialDNAcytbXUYu1ZHONGLiqiang12LIXiaoxuan1ZHUXihe1WANGChonghua1＆SHIYangbai11FreshwaterFisheriesＲesearchInstituteofJiangsuProvinceNanjing210017P．Ｒ．China2KeyLaboratoryofFishHealthandNutritionofZhejiangProvinceZhejiangInstituteofFreshwaterFisheriesHuzhou313001P．Ｒ．ChinaAbstractThemtDNAcytbgeneofOdontobutispotamophilafromfivelakeswasamplifiedandsequenced．ＲesultshowedthattheaveragecontentofA+T55．8%wasalittlehigherthanthatofG+C44．2%inthealignedsequencesof1141bp．806varia-blesitesweredetectedand87haplotypeswererecovered．Thehaplotypesdiversityandnucleotidediversitywere0．969±0．012and0．20081±0．00742respectively．ThegeneticdistancebetweenLakeTaihupopulationandLakeDazonghupopulationdis-playedthelowestupto0．137whilethatbetweenLakeChaohupopulationandLakeDazonghupopulationwasthehighestto0．424．Thefixationindicesofanalysisofmolecularvarianceamongpopulationswas0．531ThephylogenetictreeofcytbbuiltwithMEGA4．1showedthatfivepopulationswereclusteredintocladesbasedongeneticdistance．Therewaslittlegeneralflowbe-tweenO．potamophilapopulationsfromfivelakes．KeywordsOdontobutispotamophilaMitochondrialDNAcytbgeneticvariationOdontobutispotamophilaPerciformes、Gobioidei、Eleotridae、Odontobutis、1．、、、、2．．＊＊＊BE2011331、D2013-1ZJK2013132．2014-05-122014-11-06．1986～[e-mail][e-mail]．8-9cytb．1012S16SrＲNA．11ISSＲ22．12D-loop5．、、、、5mtDNAcytb55．11．1Odontobutispotamophila5、14℃．1．2DNADNASambrook13．DNA、UnicoUV-4802H1%．DNA100ng/μl－20℃．1．3PCＲcytbL14724H1591514L147245'-GACTTGAAAAAC-CACCGTTG-3'H159155'-CTCCGATCTCCGGATTACAAGAC-3'．PCＲ50μlDNA50ng2×PCＲMix25μlTaq2．5UdNTPs10μmolMgCl20．1mmol、10μmol/L1μl．94℃2min94℃45s60℃1min72℃1min3572℃10min．PCＲ．1．4DNABioEdit7．0．515MEGA4．116．DnaSP17、h、KπArlequin3．0118Kimura2-Parameters5AMOVAFst．22．1Bioedit5115mtDNAcytb1141bpGenBankKM924174～KM924288MEGA4．1115T28．1%、C28．8%、A27．7%、G15．4%A+T55．8%G+C44．2%．DnaSP80670．64%single-5Odontobutispotamophilab695tonsites163parsimonyinformativesites643344CG．11587．5h0．969±0．012π0．20081±0．007421．5hπhπ．15Tab．1ParametersummaryofhaplotypesandgeneticdiversityoffivepopulationsofO．potamophilahKπ26230．982±0．02033391．900．08573±0．0112021211．000±0．015425151．280．14788±0．0250320160．947±0．044466180．490．17472±0．0164029240．983±0．017576110．630．10679±0．025421960．468±0．14037148．160．04531±0．02485115870．969±0．012806182．940．20081±0．007422．2115mtDNAArlequinKimura2-Parameters5Fst2．0．4240．1370．237＞0．189＞0．143＞0．092＞0．056．5Fst0．5315P＜0．05．25Kimura2-ParametersFstTab．2TheaverageKimura-2-ParametersdistanceandFstamongfiveO．potamophilapopulations＊＊＊0．6580．3530．6060．7770．424＊＊＊0．2130．5330．1020．2500．276＊＊＊0．4560．2770．2950．3500．331＊＊＊0．6840．3650．1370．2060．327＊＊＊＊＊＊＊＊＊．ArlequinAMOVA5．AMOVA5．53．15%46．85%3．Gst0．07743Nm2．98115Gst0．47076Nm0．28．35Tab．3AnalysisofmolecularvarianceamongfivepopulationsofO．potamophila/%47658．40080．7143853．151107826．90371．1536746．8511415485．303151．868051002．3115mtDNAcytbMEGA4．15UPGMA1．5、696J．LakeSci．2015274、．．15UPGMAFig．1DendrogramoffivepopulationsofO．potamophilabyUPGMAmethod2．4Fu19Fs5Fs＞0Fs＜0．Fs=1．122P＞0．055、、Fs＞0Fs＜0．33．1、．5cytbT28．1%、C28．8%、A27．7%、G15．4%GGcytb20DNA21．A+T55．8%G+C44．2%9cytb．5Odontobutispotamophilab697511587mtDNA．22、23、24hπh=0．969±0．012π=0．20081±0．00742Grant252hπ．h、π26．5hπ、．．3．2AMOVA53．15%46．85%．Fst．27．0．137～0．42428-29AMOVA．5Fst0．531530．Fst．Nm2．98Gst0．07743115Nm0．28Gst0．470765．31．11ISSＲ22．1012S16SrＲNA12S、16SrＲNA12S、16SrＲNA、．5UPGMA5．mtDNAcytbSSＲ、AFLP、SNP．、、．、20082010．mtDNAcytb、．5、．、．698J．LakeSci．201527441．．19932152-61．2．、、．19965275-82．3BrownWM．EvolutionofanimalmitochondrialDNA．InNeiMKoehnＲKeds．Evolutionofgenesandproteins．Sinau-erSun-derlandMA198362-88．4SacconeCPesoleGSbisaE．ThemainregulatoryregionofmammalianmitochondrialDNAstructure-functionmodelandevolutionarypattern．JournalofMolecularEvolution199133183-91．5．b．2005322145-154．6．DNAb．2006304380-385．7．b．2006184425-430．8．OdontobutispotamophilabCytb．2010222-26．9．bCytb．201019288-290．10．12S16SrＲNA．2012401248-51．11．ISSＲ．2010382606-608．12．D-loop5．20133511-7．13SambrookJＲussellDW．MolercularcloningAlaboratorymamual3rdedition．NewYorkColdSpringHarkborLa-broatoryPress2002463-469．14XiaoWHZhangYPLiuHZ．MolecularsystematicsofXenocyprinaeTeleosteiCyprinidaeTaxonomybiogeographyandcoevolutionofaspecialgrouprestrictedinEastAsia．MolecularPhylogeneticsandEvolution2001182163-173．15HallT．BioEditv．7．0．5．Biologicalsequencealignmenteditorforwindows．IbisTherapeuticsadivisionofIsispharma-ceuticals．http//．mbio．nesu．edu/bioefit．html2005．16TamuraKDudleyJNeiMetal．MEGA4MolecularevolutionarygeneticsanalysisMEGAsoftwareversion4．0．Mo-lecularBiologyandEvolution20072481596-1599．17ＲozasJAnchez-DelBarrioJCEsseguerXMetal．DnaSPDNApolymorphismanalysesbythecoalescentandothermethods．Bioinformatics2003