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时间:2020-06-03
《利用RNAi技术抗玉米矮花叶病效果研究.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、河南农业科学,2014,43(4):69—74JournalofHenanAgricuhuralSciences利用RNAi技术抗玉米矮花叶病效果研究张莹莹,吴连成,库利霞,张赛赛,安允权,崔新建,陈彦惠(河南农业大学农学院/河南粮食作物协同创新中心,河南郑州450002)摘要:以采用RNAi技术获得的HiⅡ、HiⅡA、HiⅡB和H99抗玉米矮花叶病转基因T和Tz代为材料,通过病毒接种、草丁膦涂抹、目的基因和标记基因PCR扩增,研究了利用RNAi原理构建的目的基因对玉米矮花叶病的抗性效果以及玉米转基因后代群体鉴定筛选技术。结果表明,
2、9O.00的转基因株系抗病性超过非转基因株系,30.00的转基因株系抗病株率超过了抗病对照黄早4,转基因株系的抗病性明显提高,说明利用RNAi技术选育抗矮花叶病转基因玉米株系是可行的。4种筛选鉴定方法比较结果表明,目的基因PCR和病毒接种鉴定方法最可靠,在转基因后代群体鉴定过程中,可在苗期用200mg/L的草丁膦筛选阳性植株基础上,再采用目的基因PCR鉴定,筛选出抗性转基因植株或株系,不仅可以减少鉴定的工作量,而且可以提高鉴定的准确性。关键词:RNAi技术;玉米;矮花叶病;转基因;鉴定方法中图分类号:$513文献标志码:A文章编号:
3、1004—3268(2014)04—0069一O6EffectofMaizeDwarfMosaicDiseaseResistancebyRNAiTechnologyZHANGYing~ying,WULian—cheng,KULi—xia,ZHANGSai—sai,ANYun—quan,CUIXin-jian,CHENYan—hui(CollaborativeInnovationCenterofHenanGrainCrops,AgronomyCollegeofHenanAgriculturalUniversity,Zhengzhou4
4、50002,China)Abstract:Inthepaper,fourT1orT2generationmaterialsoftransgenicmaizeresistanttodwarfmosaicdiseasewereobtainedwithRNAitechnology,whichwereusedtostudytheresistanteffectoftargetgeneconstructedbyRNAiprincipletomaizedwarfmosaicdiseaseandthetechniquestoidentifyands
5、creentransgenicprogenypopulationthroughinoculatingvirus。daubingPPTandPCRamplificationofthetargetgeneandmarkergene.Theresultsshowedthat90.00oftransgenicplantshadhigherresistancethannon—transgenicplants,and30.00oftransgenicplantswereevenbetterthanCK(Huangzao4).Theincreas
6、eofdiseaseresistanceintransgeniclinesindicatedthatitwasfeasibletobreedthetransgenicplantswithresistancetodwarfmosaicdiseasebyRNAitechnique.ThecomparisonamongthefouridentifyingandscreeningmethodsshowedthatPCRoftargetgeneandvirusinoculationwerethemostreliable.Duringtheid
7、entifyingandscreeningprocedureoftransgenicprogenies,thepositiveplantsshouldbefirstselectedatseedlingstagewith200mg/LofPPT.fromwhichtheresistanttransgenicplantsorlineswereconfirmedusingthetargetgenePCRidentificationinordertoreducetheworkloadofidentificationandimprovethe
8、accuracyofidentification.Keywords:RNAitechnology;maize;dwarfmosaicdisease;transgene;identificationmethods收稿日期:2013—11
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