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时间:2020-04-24
《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》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
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
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