含双病原物诱导启动子无选择标记转基因烟草的获得-论文.pdf

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1、植物病理学报ACTAPHYTOPATHOLOGICAS／N／CA44(4)：433—437(2014)doi：10．13926／j．cnki．apps．2014．04．013含双病原物诱导启动子无选择标记转基因烟草的获得孔维文，王丹丹，刘爱新(扬州大学园艺与植物保护学院，扬州225009；山东农业大学植物保护学院，泰安271018)Acquirementofmarker—-freetransgenictobaccoplantscarryingtwopathogen·-indu·-ciblepromotersKONGWei—wen，WANGDan．dan．LIUAi—xin。(Schoo

2、iofHorticultureandPlantProtection，YangzhouUniversity，Yangzhou225009，China；。DepartmentofPlantProtection，ShandongAgriculturalUniversity，Tai’an271018，China)Abstract：Transgenicplantsarecontroversialfortheiredibleandenvironmentalsecurity．Marker-freetransgenicplantsCanbeproducedbytheconstructionandtra

3、nsformationofplantexpressionvectorscarryingtwinT—DNAs．TheconstructionofplantexpressionvectorsharboringtwinT--DNAsandtwopathogen--induciblepromoterswaspreviouslyreported．Thesevectorplasmidswereintroducedintotobaccoplantsandthetransgenictobaccoplantswereobtained．InthispaperwereportthatTltransgenic

4、tobaccoseedlingswereproducedthroughclassicalgenet-icsapproach．Analysisoftheseedlingsdemonstratedthatsomeofthemweremarker—freelines．ThesegregationofexogenousgenesinT1transgenictobaccoseedlingswastestedbyusingtwomethods．Firstly，theabilityofre—sistancetokanamycinwasanalyzedinTltransgenictobaccoseed

5、lingsfrom14transgenicplantlines．ItwasfoundthatthesegregationratioofⅣPnIgenesmetwellwimMendel’Slawin13transgenictobaccolines．Soitwasdeducedthat^7P7][Igenewasasasinglecopyintegratedintooneofthehomologouschromosomes．ThentheNPTIIgenesanduidAgenesof130Tltransgenicseedlingsweredetectedfromtheabove13to

6、baccolines．TheresultsshowedthatuidAgeneswereonlydetectedin20．77％oftheseedlings．NPTIIgenesweresolelydetectedin22．31％oftheseedlings．butbothexogenousgeneswerein53．85％oftheseedlings．Thesegregationratioofthetwogeneswasconsistentwiththelawofindependentassortment(9：3：3：1)．Theseresultssuggestedthatthese

7、lectivemarkergenehadnolinkagewiththereportergeneandtheyweresegregatedindependentlyintheT1transgenictobaccoplants．Thismethod，ascomparedwithtraditionalbackcross，isconfn'medamoreeasyandrapidwaytoeliminatetheantibioticresistance