分子生物技术在贝类鉴定和分类上的应用

18320059()JOURNALOFNINGBOUNIVERSITY(NSEE)Vo.l18No.3Sept.2005:100125132(2005)0320404206,(,315211):2004-12-14.:(1981-),,,,.:))),,DNA,,.,PCR(SimpleSequenceRepeat-AnchoredPCRAmplification,SSR-PCR)DNA(RandomAmplifiedPolymorphicDNA,RAPD).DNA.,,,.:;;;:Q753:AApplicationofMolecularBiologyTechnologyinDiscriminationandTaxonomyofMolluscaZHANGAi2ju,YOUZhong2jie(FacultyofLifeScienceandBiotechnology,NingboUniversity,Ningbo315211,China)Abstract:Therearesomedrawbackswhenusingtraditionalmethodstodealwiththemollusca.taxonomy,suchasmorphology,anatomy,physiology.Asanassistantmeans,molecularbiologytechnologycanexploretaxonomybasedonDNAandsettleproblemsthatcannotbesolvedbymorphology.Thus,itcanbeusedforsystematicclassifica2tionofspecies.Atpresen,ttherearemanybiologicaltechnologies,suchasallozymeRFLPRAPDSSR2PCR,etc.ThispaperfocusesontheappliedsituationofsomemolecularbiologicaltechnologiesinthediscriminationandtaxonomyofMolluscaintwoaspects,.ie,proteintechnologyandDNAtechnology.Althoughtherearesomein2consistencesbetweenresultsfrommorphologyandbiologytechnology,theimprovementsoftechnologiesandaccu2mulationoftheoriesandpracticesmakeusbelivethattheprospectofusingmolecularmethordsandmorphologyto2getherwillbemuchpromising.Keywords:molecularbiologytechnology;Mollusca;discrimination;taxonomyCLCnumber:Q753Documentcode:A(384~322).,,,.,,.2070,,DNA,,,,..,,PCR(SimpleSequenceRepeat2AnchoredPCRAmplifi2cation,SSR-PCR)DNA(Ran2domamplifiedpolymorphicDNA,RAPD),.1,2,.Davis[1],,,,,.2080,SDS.,.[2](PAGE)(Solengouldii,Conrad)(Solengradis,Dunke)(Sinon2ovaculaconstricta,Lamarck)(Cultellusattenu2atus,Dunder),,.[3](Parafossaruluseximius)(Parafossarulusstriatulus)(Arocinmalongicornis)(Stenothyraglabra),(Bellamyapurificata)(Semisulcospiracancellata),,,.,.,.[4][5]A-,(),,2.[6](EST)(MDH),[7],Davis[8].,[9],25(Triculacristella),,.,,,.,[10,11]..4053:2DNA,DNAPCR.PCR(Polymer2aseChainReaction,PCR),DNA,DNA,.,.2.1DNA.DNA,,,.DNA.16SrRNACOI,;18S~28SITSDNA,,.ITS.2.1.1ChristinaMSpolsky[12]3b,Davis[8]3.Davis[13]2HydrobiodPomatiop2sidae221(COI),COI.Monica[14]3(12S,16SCOI),,(Aplysiidae).BarguesMD[15]13166ITS-2,ITS-2.Annamaria[16]patellacaerulea,patellaaspera,pa2tellarusticamtDNACOI,,.,.Emberton[17]Rosenerg[18]rRNA,rRNA,.Schilthuizen[19]5ITSI,.2.1.2.Banks[20]16SrDNA,2(CrassostreagigasC.sikamea).An2derson[21]ITS,(Sac2costreacommercialis,IredaleetRoughley)(S.glomer2ate,Gould).Que[22]ITS,,ITS.Canapa[23]mtDNA16SrRNA,.[24]16SrRNAPCR,16SrRNA,16SrRNA.Marco[25]162312S16SrRNA,.[26](Crassostreasp.)(Amussiumpleurondect2es,Linnaeus)rRNAITS,ITS.Adriana[27](Veneroidea)82918SrRNA,,Myoida,2.[28]8(Pinctadamartensii)ITSI,ITSI,,ITS-1,406()2005.2.2PCR2RFLPPCRDNA,RFLP.RFLP,PCR-RFLPDNA,.Caldeira[29]PCR-RFLP:Biomphalariastraminea,B.intermedia,B.kuhni2ana,B.peregrina.2,,DNA,.2.3RAPDDNAPCRDNA.RAPD,DNA,,RAPD,.RAPD,.RAPDPCR,,,.Juliettelangand[30]RAPD(Bulinus)3.Abdel2hamid[31],,,RAPD.RAPD.[32](Crassostreatalienwhanensis)(Crassostreaplicatula)3,3(Crssostrea),,.[33]RAPD(Pinctadachemmitri),RAPD,2,2.2.4(microsatellitemarkers)2~6.,PCR,DNA,.Jones[34].Inmaculada[35](Ostreaedulis,O.stenti2na,Crassostreaangulata,C.gigas,C.gasar,C.ariaken2sis,C.virginicaC.sikamea)DNA,O.edulisO.stentina,C.angulataC.gigas..,Bulinustruncates,Bu2linusterrestris,MelanoidestuberculatBiomphalariaglabrata.DNA,,DNA,.2.5PCR(SSR2PCR)PCRDNA.SSR2PCR,,.RAPD,,,,.3,,,,,,.,,,,,,4073:.,,,,,.:[1]DavisGM,LindsayGK.Discelectrophoreticanalysisofmolluscanindividualsandpopulations[J].Malacolo2gia,1967,5:88-111.[2],,.[J].(),2000,18(3):40~43.[3],.[J].(),2002,27(1):200~204.[4],,.[J].,1980,2(3):206~208[5].[M].:,1990.[6],,.[J].,1999,12(1):52~55.[7],,,.[J].1989,24(1):7~9.[8]DavisGM,ZhangY,GuoYH,eta.lPopulationge2neticsandsystematicstatusofOncomelaniahupensis(Gastropoda:Pomatiaopsidae)throughoutChina[J].Malacologia,1995,37(1):133-156.[9],,,.5[J].,1995,16(3):189~192.[10].[J].,1992,27(4):15~17.[11],,,.[J].,2003,22(2):173~179.[12]ChistinaMS,DavisGM,ZhangY.Sequencingmeth2odologyandphylogeneticanalysis:cytochromebgenesequencerevealssignificantdiversityinChinesepopula2tionsofOncomelania(Gastropoda:Pomatiopsidae)[J].Malacologia,1996,38(1-2):213-221.[13]DavisGM,WilkeT,SpolskyC,eta.lCytochromeoxi2dase12basedphylogeneticrelationshipsamongthePoma2tiopsidade,Hydrobiidae,RissoidaeandTruncatellidae(Gastropoda:Canogastropoda:Rissoacea)[J].Mala2cologia,1998;40(1-2):251-266.[14]MonicaMedina,PatrickJWalsh.MolecularSystematicsofthe=rderAnaspideabasedonmitochondrialDNAse2quences(12S,16S,andCOI)[J].MolecularPhyloge2neticsandEvolution,2000,15(1):41-58.[15]BarguesMD,VigoM,HorakP,eta.lEuropeanLym2naeidae(Mollusca:Gastropoda),intermediatehostsoftrematodiases,basedonnuclearribosomalDNAITS22sequences[J].Infection,GeneticsandEvolution,2001(1):85-107.[16]AnnamariaMauro,MarcoArculeo,NicoloParrinello.MorphologicalandmoleculartoolsinidentifyingtheMediterraneanLimpetsPatellacaerulea,PatellaasperaandPatellarustica[J].ExperimentalMarineBiologyandEcology,2003,295:131-143.[17]EmbertonKN,KuncioGS,DavisGM,eta.lCom2parisonofrecentclassificationsofstylommatophoranland2snailfamilies,andevaluationoflarge2r