48个油橄榄品种的遗传多样性及聚类分析(英文)

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1、12162013opaeaL．frOmtheolivecultivarre-electrophoresisunder4V／cmwithStatisticsandgeneticanalysissourcenurserygardenofYongren0．5XTBEaseIectrODhOresjsbufer．ThepresenceandmolecularweightofCountyinChuxiongYiAutonomousElectrophoresisresultswereobservedamplifiedbandsinelectropherogramsPrefecture

2、ofYunnanProvince，whichandphotographedwiththeultravioletwasdeterminedaccordingtoDNAprovidedbasisforstandardizedman-gelimagingsystem。Marker：electrophoresisresultswithagementandfurtherdevelopment。uti-Table1Origins，sourcesandmain‘usagesof48OleaeuyopaeaL．cultivarsIizationandbreedingoftheseOlea

3、euyopaeaL．germplasmresources．MaterialsandMethodsMaterialsTotally480／eaeuyopaeaL．culti—varsplantedintheolivecultivarre-sourcenurserygardenofYongrenCountyinChuxiongYiAutonomousPrefectureofYunnanProvincewerecollectedasexperimentalmaterials(Table1)。including35introducedculti-varsfNo．1—35)and1

4、3nativecultivarsfNo．36—48)．TenderOleaeuyopaeaL．1eaveswerecollected．putintoziplockbagcontainingsilicagel，broughtbacktotheIaboratory．andpreservedjn一20oCrefrigeratorbeforeextractionofge-nomicDNA．MethodsGenomicDNAextractionandquali-tydetectionGenomicDNAof48OleaeuyopaeaL．cultivarswasex—tracted

5、usingplantgenomicDNAex—tractionkitfTiangen)．ExtractedDNAwasdetectedby0．8％agarosegelelectrophoresisinaccordancewiththepreviousliteraturet徊．ISSR-PCRamplificationThetotaIlSSR-PCRreactionvolumewas2OuI，containing1XTa口Buffer，3．5mmOI／LMg“。0．4mmOl／LdNTPs．1．0iJmol／Lprimers．1．0UOfqDNApoly—meraseand

6、20ngofDNAtempiate．ThelSSR．PCRamplificationwasstartedwithinitiaIdenaturationat94oCfor5min，followedby40cyclesofde—nalurationat94℃for30S．annealingat50-55oCfor30S，andextensionat72℃for120S：theamplificationwascompletedbyholdingthereactionmixtureat72℃for10mintoallowcompleteextensionofPCRproducts

7、．PCRproductswerestoredat4oC．Samplesof48OleaeuyopaeaL．cul-tivarswereamplifiedusing11screenedprimerswithhighdefinition，stabilityandpolymorphism(synthe-sizedbyBeijingAugctBiotechnologyCO．，Ltd．)．SequencesandannealingtemperaturesofeachprimerwereshowninTable2．PCRproductsw