中药材鳖甲的位点特异性PCR鉴定研究

PCR,,a(,210097):DNA12SrRNA,,1,DNA,PCR,10,3,DNA12SrRNA:;;PCR,:R282174013:A:02532670(2001)08073603AuthenticationofTCMCarapaxTrionycisbyallele-specificdiagnosticpolymeraschainreactionLIUZhong2quan,WANGYi2quan,ZHOUKai2ya(InstituteofGeneticResources,NanjingNormalUniversity,NangjingJiangsu210097,China)Abstract:ObjectTodevelopaconvenientandpracticalmethodfortheidentificationofCarapaxTrionycis1MethodsBasedonthesequencevariationsof12SrRNAgenebetweenPelodiscussinensisandothersoftshellturtles,apairofallele2specificprimerswasdesignedtodistinguishP.sinensisfromotherspeciesofTrionychidae.DNAwereextractedandanplifiedandCarapaxTrionyciscouldbeidentifiedaccu2ratelybypolymerasechainreaction(PCR)usingtheprimers1ResultsTensamplesofturtleshellfromdifferentsourceswereindentifiedbytheallele2specificPCRwiththeprimers1TheresultindicatedthatthreesamplesweresubstitutesofCarapaxTrionycis,consilientwiththeresultfromDNAsequenceanaly2sis1Themitochondrial12SrRNAgenefragmentofP.maculatusandafakedimitationhadalsobeense-quenced1ConclusionTheprimerscouldbeusedaskeycomponentsinCarapaxTrionycisidentificationkit1Keywords:CarapaxTrionycis;allele2specificprimer;diagnosticpolymerasechainreaction(PCR),1995TrionyxsinensisWiegmann[1](TrionyxsinensisPelodiscussinensis[2]),,,Peleasteindachneri(Siebenrock)Lissemyspunctata(Bonnaterre)Pe2lochelysbibroni(Owen)[3,4],,,[3],,DNA500bpCytb[5];DNA,110bp12SrRNA,,2DNA[6],DNA,,12SrRNA,,,,1637ChineseTraditionalandHerbalDrugs2001328a:2000211210:(No139870913)(98KJB360010)333:(19632),,,,PlantaMedica10©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.,DNA,PCR,1111:44,,Pelochelysmacu2latus(Heude),,110,4(JP12JP4);6(JP52JP10)14Pelodiscussinensis[2]Pi4,PaleasteindachneriPt2PelochelysmaculatusPm1LissemyspunctataLpGenbank112DNA:DNA,SDSö,,DNA[7,12]113PCR:PCR[7]ABI310BigDyeDNAPEAppliedBiosys2tems,114:ClustalWDNA,MEGADNA,IT2L02IT2H02(),170bp12SrRNAL1091H1478115PCR:PCREB,DGS27600(UVP,Inc1)2211:L1091H1478DNA,GenBank,AF321281,(JP2)[8],GenBank(U81337),405bp,1616192%32163%JP2(Pi)914%;(Pt)8162%;(Pm)3213%;(Lp)16138%(2)2512SrRNAö()()(%)OTU123451Pi19ö864ö5529ö2223ö132Pt619267ö5735ö1622ö113Pm311483216358ö4964ö564Lp14141141413111034ö245JP2914081623213516138212PCRA2L1091H1478,55B2IT2L02IT2H02,55N2M2Marker,DL2200011PCR21211PCR:,55,DNAL1091H1478DNA,400bpDNA(12A),(Pi)170bpDNA,(Pt)(Pm)(12B)21212PCR:55,,10(2),,1,510,24(2)C2L1091H1478,55D2IT2L02IT2H02,55P2N2M2Marker,DL2200013PCR310PCR,4,3[6](JP2),GenBankBLAST,,,,737ChineseTraditionalandHerbalDrugs2001328©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.,6,,[9],,44JP2,10,IT2L02IT2H02Tm,IT2L0272,IT2H0268,,DNA,PCR,,[1012],:[1][M]1199511[2],,1[M]111111:,19981[3],,,1[J]1,1996,21(9):51825221[4]1[J]1,1998,11(1):321[5],,,1DNA[J]1,1996,31(6):47224761[6],,,1[J]1,1998,29(1):282301[7],,1DNA[J]1,2000,31(5):34323451[8],,112SrRNA[J]1,1999,45(3):26022671[9],,,1[M]1:,19981[10],,,1CytbPCR[J]1,1998,33(12):94129471[11]WangYQ,ZhouKY,XuLS,etal1Authenticationofananimalcrudedrug,Zaocys,bydiagnosticPCR[J]1BiolPharmBull,2000,23(5):58525881[12],,,1PCR[J]1,1999,34(12):94129451,,,a(,710062):,,,,MS,BA012mgöL+NAA110115mgöL;BA1mgöL+NAA011012mgöL+GA368mgöL+CH300mgöL;BA1mgöL+NAA011mgöL+GA32mgöL;IBA011mgöL+NAA015mgöL:;;;:R282113:B:02532670(2001)08073804EstablishmentofplantletregenerationsystemofRubusidaeusXIAOYa2ping,WANGZhe2zhi,ZHANGZhi2qin,LIUQuan2hong(CollegeofLifeScience,ShanxiNormalUniversity,XianShanxi710062,China)Abstract:ObjectToestablishaplantletregenerationsystemofRubusidaeusL1forthepurposetoobtainalargenumberofhighqualityseedlinginashorttime1MethodsStemapexandpartofthestemwereusedastheexplantandtheoptimalculturemediaandconditionswereselectedbyorthogonaldesign1ResultsAnoptimumculturemediumfortheinductionofcallus,adventitiousbudandrootwasobtainedwhichcanbecarriedoutinthelaboratorywithcomparativeeaseandgoodrepeatability1ConclusionAbasicmedium+BA012mgöL+NAA110115mgöLwasmostsuitablefortheinductionofcallus;a837ChineseTraditionalandHerbalDrugs2001328a:2001202208:(19562),,19812,3,12,2,20©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.