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1、中国水稻科学(ChinJRiceSci),2016,30(3):256-264http://www.ricesci.cn256DOI:10.16819/j.1001G7216.2016.5190转Cry30Fa1基因抗褐飞虱水稻的获得及鉴定王海鹏黄晓西梁越洋朱军张翠霞王秀梅贡常委郑爱萍邓其明李双成王玲霞李平王世全∗(四川农业大学水稻研究所,四川温江611130;∗通讯联系人,EGmail:sqwangscau@163.com)DevelopmentandIdentificationofInsectResistantTransgenicRicewithCry30
2、Fa1GeneWANGHaiGpeng,HUANGXiaoGxi,LIANGYueGyang,ZHUJun,ZHANGCuiGxia,WANGXiuGmei,GONGChangGwei,ZHENGAiGping,DENGQiGming,LIShuangGcheng,WANGLingGxia,LIPing,WANGShiGquan∗(RiceResearchInstitute,SichuanAgriculturalUniversity,Wenjiang611130,China;∗Correspondingauthor,EGmail:sqwangscau@163c
3、.om)WANGHaipeng,HUANGXiaoxi,LIANGYueyang,etal.DevelopmentandidentificationofinsectresistanttransgenicricewithCry30Fa1gene.ChinJRiceSci,2016,30(3):256G264.Abstract:ModifiedCry30Fa1,aBacillusthuringiensis(Bt)geneencodingahighGefficiencyinsecticidalproteintothebrownplanthopper(BPH),was
4、introducedintoShuhui818(R818)byAgrobacteriumGmediatedgenetictransformationand46transgenelineswereobtained.WeconfirmedthevalidtransformationbyRTGqRCRandWesternblotattranscriptionallevelandtranslationallevel,andfixedthestableexpressionCry30Fa1inhomozygosis.Theclassicalpedigreebreeding
5、methodswerealsousedtoselecttheimportantagronomictraitsintransgeniclines.Furthermore,weevaluatedtheresistancetoBPHofdifferentlinesingrowthchamberandfield,andfoundthattheR818GCry30FaldisplayedhigherresistancethanuntransformedR818inseedlingstage.WealsoobservedthereducedpopulationofBPHf
6、eedingthetransgeniclines,suggestingthatthetransformedCry30Fa1enhancedresistibilitytoBPHinrice.WecultivatednewBTproteintransgenicricerestorerlineR818GCry30Fal,whichmayprovidenewBPHresistancematerialtothreelinehybridricebreedingandenrichBPHresistancegermplasmresourcesinrice.Keywords:b
7、rownplanthopper;AgrobacteriumGmediatedmethod;insectresistance;Cry30Fa1王海鹏,黄晓西,梁越洋,等.转Cry30Fa1基因抗褐飞虱水稻的获得及鉴定.中国水稻科学,2016,30(3):256G264.摘要:将编码高效杀褐飞虱蛋白的苏云金芽胞杆菌基因Cry30Fa1密码子改造后,通过农杆菌介导法转入蜀恢818(R818),并最终获得46个转基因植株.通过定量PCR及WesternBlot鉴定了Cry30Fa1在转录水平和蛋白水平的表达,并通过分子检测固定稳定表达的抗性基因,并结合传统育种系谱法选
8、择具有优良农艺性状株系.对选育的株系在