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1、甘蔗ROC16的转基因研究GenetransfertosugarcaneROC16摘要甘蔗(SaccharumofficenarumL.)是我国最主要的糖类经济作物,其生产率的大小将直接影响到蔗农的收入和国民经济的稳定发展。但是在实际的甘蔗生产过程中,各种病虫害的侵害将大大降低甘蔗的产量和品质。在众多的病虫害中,常见的甘蔗害虫是甘蔗螟虫(PiatraeaSaacharalisF.)也是最主要的虫害。本研究在于将修饰的豇豆胰蛋白酶抑制剂基因(sck)及苏云金杆菌毒蛋白基因(Bt)导入甘蔗ROC16种
2、质中。探索获得了农杆菌介导ROC16转基因研究的相关科学数据:诱导甘蔗ROC16愈伤组织的最佳2,4-D浓度为2.5mg/L(诱导率为91.7%)介于ROC16愈伤组织会因2,4-D浓度过高而不易诱导芽分化,在实际的愈伤诱导中,采用了1mg/L(诱导率为72.9%)。采用2mg/L6-BA+1mg/LKT+200mg/Lcar进行芽的高浓度培养基中诱导刺激分化15天,再转入0.5mg/L6-BA+0.25mg/LKT+200mg/Lcar的低浓度培养基中15天,最后转入1mg/L6-BA+0.5m
3、g/LKT+200mg/Lcar培养基中持续分化,可解决ROC16在常规浓度下(1mg/L6-BA+0.5mg/LKT+200mg/Lcar)难以诱导芽分化问题。关键词:甘蔗;ROC16;农杆菌;转基因;sck基因;Bt基因ABSTRACTThesugarcaneisthemostmaincarbohydrateindustrialcropinmycountry,Itsproductivitywilldirectlyaffectthesizeofnationaleconomicincome/sta
4、bledevelopment,Butinactualsugarcaneproductionprocess,variousinsectviolationswillgreatlyreducethesugarcaneyieldandquality,Inmanyoftheinsectpest,commoncanecaneisstemborerandoneofthemostimportantpests.ThisresearchistomodifycowpeatrypsininhibitorSCKgenean
5、dgeneROC16sugarcaneBtgermplasm.TheexplorationobtainedtheagriculturalbacillustoliebetweenleadstheROC16transgeneresearchtherelatedscientificdata:InductionsugarcaneROC16injuriestheorganizationthebest2,4-Ddensityis2.5mg/L(inductivityis91.7%),Roc16betweenc
6、allusbecause2,4-dhighconcentrationandeasilyinducedbudpointsactualinjuriesintheinduction,hasused1mg/L(inductivityis72.9%).Using2mg/L6-BA+1mg/LKT/caron200mg+highconcentrationsofbuddifferentiationmediuminducedstimulusfor15days,Againinto0.5mg/L6-BA+0.25mg
7、/LKT200mg+/caroflowconcentrationmediumfor15days,Finally1mg/Lto6-BA+0.5mg/LKT200mg+/carmediumcontinuousdifferentiation.ROC16cansolveinconventionalconcentration(1mg/L6-BA+0.5mg/LKT/car)200mg+toinducebuddifferentiationquestion.ButafterROC16genetictransfo
8、rmationtoprovidethescientificbasis.Keywords:Sugarcane,ROC16,Agrobacterium-mediated,transgenic,sckgene,Btgene目录1前言11.1甘蔗概况11.2甘蔗遗传转化国内外研究进展12材料与方法32.1材料32.1.1植物材料32.1.2菌种和质粒32.1.3试剂32.1.4主要仪器设备32.2方法32.2.1技术路线32.2.2cpti基因的修饰42.2.3植物表达载体的构建42.2.