甘藍型油菜顯性核不育基因的AFLP標記及品種(系)的SSR指紋圖譜研究.pdf_第1頁
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1、中國農(nóng)業(yè)科學院碩士學位論文甘藍型油菜顯性核不育基因的AFLP標記及品種(系)的SSR指紋圖譜研究姓名:程勇申請學位級別:碩士專業(yè):作物遺傳育種指導教師:李云昌20050620AbstractRapeseedisoneofthemajoroilsecdcropsinChinaBreedersstillfacethecomlllOnandmaingoalofimprovingtheproductionlevelofrapeseedLieta

2、l(1985)hadproposedforthefirsttimethatthesterilityofdominantgeniemalesterility(GMS)followedadigenicmodeofinheritancewithepistaticinteraction,andestablishedtwolinesystemandthreelinesystemasGMSbreedingmodelsThishypothesisan

3、dmodelsmadeheterosisutilizationofGMStobeaprimarystrategyinBrassicanapusLbreedingThedominantGMSisanexcellentsysteminhybridbreedingforitsstableandcompletesterilityimpervioustoenvironment,vigomuslygrowthandeasytotransformst

4、erileandfertilegenesInthreelinesystem,homozygousGMStwotypelineisafoundationmaterialselectedfromhybridofrestorerandsterileplantofheterozygoustwotypelineTheselectiondependsonanalyzingofsegregationratebetweenfertileandsteri

5、lehybridsTomakethissystemawideruseitisnecessarytodevelopmolecularmakersofitssterilegenesandanalyzethediversityofparentallinestomakethemostoptimizeshybridsThepresentstudyincludes(1)breedingofahomozygousGMStwotypelinefrom且

6、napusheterozygoustwotypeline70056ABandarestorerline9548;(2)developingA砌(AmplifiedFragmentLengthPolymorphism)molecularmarkersofmalesterilitygenebasedOil70056ABsib—matingpopulationbyBSA(BulkedSegregantAnalysis)metIlod;(3)a

7、nalyzingtheSSRfmgefprintofparentallinesincludingrestorer9548inordertogainthegeneticdistancedataforbreedingpmgramMainresultsofthepresentstudiesareasfollowing:1ThegenotypeofrestorerlinewasidentifiedasmsmsR/瞬faftercrossedst

8、erileplantofheterozygonstwotype70056ABwithrestorer9548andinvestigatedthesterilityofF1,F(xiàn)2andF3populationsTheinvestigationresultsshowedthatF1plantswere100%fertile,F(xiàn)2populationsweretestedtofitthesegregationrateof1:1by。2test

9、andF3populationhadtheidealfertilitysegregationTheseresultswouldleadtosuccessfulselectionofhomozygousGMStwotypelineinthenearfuture2Fertilebulk(A)andsterilebulk@wereconstructedinthesib—matingpopulation70056AB(70056A70056B)

10、byBSAmethodandwereusedinthefoUowingoptimizationTheprotocolofAFLPmolecularmarkerapproachWasoptimizedinrestrictionendonucleasesdigestion,ligationanddilutionoftemplatesThetwo_stepmethod,ieseparatinlngthedigestionandligation

11、steps,WaseonfLrmedtobebetterforthedigestionactivityofrestrictionenzymesandresultinlegibleandrepeatablebandsThetemplatedilutionwasnotanimportantparameterintwostepmethodandhadlittleinfluencetoamplificationresultsWhereasino

12、nestepmethod,maindifferentialfragmentsappearedinthesamesampleonlywithdifferentdilutiongradesoftemplates,whichmisledtheprimerscreeningresultAnoptimizedAFLPprotocolwerethenconfirmedasfollowing:genomicDNAwereincubatedfor5hr

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