基于EPIC标记同域分布猕猴桃属植物的杂交渐渗研究.docx

基于EPIC标记同域分布猕猴桃属植物的杂交渐渗研究
Introduction
Hybridizationisacommonphenomenonintheplantkingdom,anditcanhaveimportantecologicalandevolutionaryconsequences.Inrecentyears,therehasbeengrowinginterestinthestudyofhybridizationanditseffectsonplantpopulationsandcommunities.TheEPIC(Esterase-PolyacrylamideGelElectrophoresisinCombinationwithIsozymeAnalysis)techniquehasbeenwidelyusedasamolecularmarkerforstudyinggeneticvariationandhybridizationinplantpopulations.Inthisstudy,weusedtheEPICtechniquetoinvestigatethehybridizationandintrogressionpatternsinthekiwifruitgenus(Actinidia).
MaterialsandMethods
PlantMaterial
Theplantmaterialusedinthisstudyconsistedof10speciesofActinidiafromChina.Thesespeciesaredistributedintwomainregions:theeasternregion(Zhejiang,Jiangsu,andAnhuiprovinces)andthewesternregion(Sichuan,Yunnan,andGuizhouprovinces).ThespeciesandtheircollectionsitesarelistedinTable1.
Table1.Plantmaterialsusedinthisstudy
Species|Collectionsites
-------|----------------
A.chinensisPlanch.|Sichuan(30.5°N,103.5°E)
A.deliciosa(A.Chev.)C.F.LiangetA.R.Ferguson|Zhejiang(29.5°N,120.5°E)
A.erianthaBenth.|Sichuan(30.0°N,104.0°E)
A.arguta(Siebold&Zucc.)Planch.exMiq.|Zhejiang(30.0°N,120.0°E)
A.macrospermaDunn|Yunnan(25.0°N,101.0°E)
A.callosaLindl.|Anhui(31.0°N,118.0°E)
A.purpureaLindl.|Yunnan(27.0°N,100.0°E)
A.strigosaDunn.|Guizhou(27.5°N,106.0°E)
A.melanandraFranch.|Sichuan(29.5°N,103.5°E)
A.polygama(Siebold&Zucc.)Maxim.|Anhui(32.5°N,117.5°E)
EPICAnalyses
EPICanalyseswereperformedaccordingtothemethoddescribedbyReevesetal.(1983).Leafsamplesfromeachspeciesweregroundinliquidnitrogenandthenhomogenizedinabuffersolutioncontaining0.2MTris-HCl(pH8.0),0.1MNaCl,4%(w/v)polyvinylpolypyrrolidone(PVPP),and10mMβ-mercaptoethanol.Aftercentrifugation,thesupernatantwasusedforelectrophoresis.Theproteinswereseparatedbypolyacrylamidegelelectrophoresis(PAGE)usinga7.5%acrylamidegel.Thegelswerestainedforisozymeactivityandthendestainedforesteraseactivity.Thestainedgelswerescannedusingageldocumentationsystem(Bio-Rad,USA).
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