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1、西北農(nóng)林科技大學(xué)碩士學(xué)位論文蘋果黑星病菌遺傳多樣性的SSR分析姓名:董艷玲申請學(xué)位級別:碩士專業(yè):植物病理學(xué)指導(dǎo)教師:楊家榮20050601GENETICDIVERSITYOFVenturiainaequalisOFAPPLEWITHSSRANALYSISABSTRACTApplescab,causedbyVenturiainaequalis(Cooke)Wint,isoneofthemostseriousdiseasesofapple

2、andornamentalcrabappleBothleavesandfruitsCallbeinfectedInfectedleavesmaydropearlyresultinginweakeningtrees,withpoorfruitproductionInfectedfruitsareblemishedandoftenseverelydeformedInChinaapplescab,whichlimitistoonlyOccur

3、inafewlocalsites,ispointedasquarantineobjective,butinrecentyears,thisdiseaseisexpandingtOlargerareasandseverelythreateningtoappleindustryinChinaItwasanewdiseasefoundedfirstlyin1997inXinping,Liquan,Yangling,andXunyiinShaa

4、nxiprovinceThemainresultsobtainedbymolecularanalysiswithSSRthchniqueareasfollows:1ThegenomicDNAofVenturiainaequalisofapplewasextractedbyuseingSquashBlotmethod,whichcarlreducetheharmoftoxicitytoexperimenters,chemicalreage

5、ntsandneedinexpensiveapparatus,ThemainadvantageofthistechniquecangreatlyspeedupexperimentandissimpleandeasytooperationManysamplescanbecrushedontoasinglemembranepiece,andasingleoperatorCananalysehundredsofsamplesinonedayI

6、tcangreatlycutshortofthetimeofextractinggenomicDNAfromVenturiainaequalisofappleUsuallytheconventionalmethodtookmorethanonemonthtoextractgenomicDNAfrommyceliumgrowninculturemedium,butbyusingthismethodonlytookseveraldays2W

7、ehavesetuptheoptimizingSSRreactionsystemofVenturiainaequalis:each25ItLamplificationreactionsolutionWascomprised10xBuffer(MgCl220mmol/L)25ItL,dNTPl00gmoL/L,primers05lamol/L,TaqDNApolymerase15U,templateDNA2mg/L,ddH20195雎L。

8、Amplicationprogramis:2minat93(2,30Sat57“(2:then40cyclesthatconsistedof1minat72℃。30Sat93℃,30Sat57℃;followedby10minextensionat72℃TheresultofPCRamplificationshowedthatthePCRsystemhadgoodrepeatandstability3Wehaveanalyzedtheg

9、eneticdiversityof160isolatesoriginedfromUK,IndiaandSharmxiprovinceofChinaResultsshowedthatisolatesodginedfromChinaUKandmostofisolatesoriginedfromIndiahadcloserelationship,butsomeIndia’SisolatesRDl,G1—10,RI1,G28,G12andRI1

10、2variedinsomedgree,andtherewerenorelationship、vi也thosefromShannxiprovinceofChinaandUKResultsalsoindicaredthatthereWasnelatetionshipbetweenthegeographicdistanceanddiffemetapplecultivarsThisstudyearlhelpUStohavebeRerunders

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