三葉斑潛蠅實(shí)時(shí)熒光PCR檢測(cè)及地理種源關(guān)系研究.pdf

1、湖南農(nóng)業(yè)大學(xué)碩士學(xué)位論文三葉斑潛蠅實(shí)時(shí)熒光PCR檢測(cè)及地理種源關(guān)系研究姓名：馮賢申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：農(nóng)業(yè)昆蟲與害蟲防治指導(dǎo)教師：馬駿陳乃中20070601AbstractLiriomyzatrifolii(Burgess)(Diptera：Agromyzidae)isallimportantpestofvegetableandcut—flowercropsaroundtheworldLiriomyzatrifoliiinChinam

2、ainlandwasfirstreportedfromZhongshanofGua／tgdongprovincein2006WereportherethatthispesthasrecentlyexpandeditsdistributioninChina，alongwithahostplantrangeextensionandpopulationexplosionThemolecularidentifyingLiriomyzatrifo

3、lii(Burgess)basedonreal—timePCRwasestablishedin[hispaperUsingpartialDNAsequencesofmitochondrialCOIofLtrifoliioccurredinGuangdongandTaiwanprovincesinChinaJapan，Philippines，Israel，GermanyUS氣MexicoandHonduras，andrelatedspec

4、iesfromGenbank，the三trifolii—specificprobedistinguishingfromLsativaeBlanchardisdesignedItobtainedthesanleresultsinindividualsofdifferentstagesItisasentitive，fastandlesscostlyassaysystemcomparedtoothermolecularmethod，sucha

5、srandomamplifiedpolymorphicDNA—PCR，PCRrestrictionfragmentlengthpolymorphism，DNAsequencingandmultiplexpolymerasechainreactionThepartialCOlsequenceofmtDNAfrom14leafminerspopulationswereanalyzedTheprobesandprimersforL吮白liiw

6、eredesignedbasisonthealignsequenceoftheamplifiedCOImtDNAofLiriomyzasppbyusingtheCLUSTSAL181ofthesequencesoftwareTheprobewaslabeledtwofluorescentdyes，thatisareporterof6carboxyfluorescein0：AM)atthe5’endandaquencherof6carbo

7、xytetramethhylrhodamine(TAMRA)atthe3’endThereal—timePCRmethoddevelopedinidentitifyingLtrifoliiinthisstudywasin4hrs，Itisastableandpreciseidenti助ngmethodTheresultsshowedthatthemitochondrialcytochromeoxidaseIgenesequencedfr

8、om14populationscollectedfromZhuhai，Ningbo，Taiwan，ZhongshaninChinaandJapan，Israel，GermanyandUSAwereidenticalNogeneticvariationwasobservedbetweenpopulationeitherfromdifferentgeographicallocalitiesorfromdifferenthostplantsT

9、hephylogeneticanalysisindicatedthatChinapopulationwasgroupedintothetrifolii—Wclade，belongingtoanonpepperlineage，whichalsoincludesotherrecentlyintroducedAsianpopulationsKeywords：Liriomyzatrifolii，mtDNACOlgene，molecularide