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

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