杉木分子細胞遺傳學研究.pdf

1、南京林業(yè)大學博士學位論文杉木分子細胞遺傳學研究姓名：徐進申請學位級別：博士專業(yè)：林木遺傳育種指導教師：施季森20060601MolecularCytogeneficStudyonChinesefir(Cunninghamialanceolata)Candidate：XuJinSupervisor：ProfessorShiJisen(DepartmentofforestrygeneticNanjingForestryUniversityN

2、anjin9210037)AbstractMolecularCytogeneticsisthedevelopmentdirectionofthemoderncytogeneticsChinesefir(Cunninghamialanceolata)，oneofthelocalandeconomicallyimportant仃eespeciesofChina，hasbeenstudiedincytologyandmolecularcyto

3、geneficslevelsbyusingfluorescencedyeing，chromosomebanding，chromosomemicrodissectionandmicrocloningandinsuithybridization(ISH)Theresultswillacceleratethetheoreticaldevelopmentofthemacroscopicalcytologyandthemicroscopicalm

4、olecularcytogeneticsofChinesefirThemitosismetaphasechromosomeswerestudiedinthispaperTheresultsshowthat6ofthe1lchromosomesofChinesefirwereeasilyidentifiedbyCbanding，CMAbandingismoresuitableforthefluorescencebandingofChine

5、sefirthanDAPIFluorescencebandingcanbedetectedinthechromosomesbyusingCMA，butnobandingbyDAPIThemeiosisruleofPMC(pollenmothercell)ofChinesefirwasstudiedbyconsecutivelysamplingandsquashingTheresultsshowthatthemeiosisphasesof

6、PMCofChinesefirhavehi曲correlationwiththecolortransformationofthemaleflowersItundergoesaseriesofchanges：filemot，emerald，green，brown，puceAccordingtotheobservationofmeiosisⅡtelophasetetradofChinesefir，themeiosisoftelxadissi

7、ngleplane，thedivisionmodeofcytoplastissimultaneousWefirstlyreportedthatthereweresomeabnormalcytologybehaviorsinthemeiosisofPMCQollenmothercell)：univalence，Inultivalencechromosomebridgeslaggingchromosomes，nonequivalentdiv

8、ision、micronucleusandetcThevigorofpolleniscommonlylowThisphenomenonhascorrelationwimthenon—equivalentdistributionofgermplasmduringmeiosisofPMC，whichresultsinthesterilityofmicrosporeWesuccessfullyisolatedthesinglechromoso

9、mewithasatelliteofChinesefir，andamplifiedthechromosomebyusingDOPPCRwi山afragmentfrom100bpto700bpThePCRproductswereligatedtopGEMTvector(Promega)ThequalityofthesinglechromosomelibrarywasscreenedbydothybridizationTheresultss

10、howedthattheproportionofsin#e／lowcopywas571％andthemi0&ighcopieswas429％WeanalyzedthemeiosisPMCandmitosisroottipsbyFISHwithribosome28SprimerTheresultsshowedthatthehybridizationsignalsweredetectedininterphaseandprophaseduri

11、ngthemitosisofroottipcellsandthemeiosisofPMCWefirstlyanalyzedthemeiosisPMCandmitosisroot卸sbyGISH麗ⅡltheprimersofJapancedarandMetasequoiagenomes，respectivelyTheresultsshowedthatthehybridizationsignalsweredetect05inmostchro