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当前位置: 首页 > 文章 > 农杆菌介导的宁麦13成熟胚遗传转化体系的构建 麦类作物学报 2012,32 (3) 415-420
Position: Home > Articles > Agrobacterium-mediated Transformation by Mature Embryo of Wheat Ningmai 13 Journal of Triticeae Crops 2012,32 (3) 415-420

农杆菌介导的宁麦13成熟胚遗传转化体系的构建

作  者：
蒋苏;马鸿翔;张旭;余桂红;孙晓波
单  位：
江苏省农业科学院江苏省农业生物学重点实验室
关键词：
小麦(Triticum aestivum);宁麦13;成熟胚;农杆菌;遗传转化
摘  要：
宁麦13是目前生产中应用面积较大的弱筋小麦品种,以此品种建立农杆菌介导的成熟胚遗传转化体系有助于对该品种的个别性状进行针对性改良。本研究利用分别携带不同质粒pCXK1301和pAt-MYB12u的农杆菌株系GV3101对宁麦13成熟胚愈伤组织进行转化,通过对预培养时间、共培养时间和干燥处理等参数的研究,建立了农杆菌介导的宁麦13成熟胚遗传转化体系。宁麦13成熟胚转化体系的条件为:利用预培养6d的宁麦13成熟胚愈伤组织作为转化受体,使用含pAtMYB12u质粒的农杆菌株系GV3101,侵染菌液浓度OD600=0.6,侵染时间60min,采用干燥共培养4～5d(23℃,暗培养),诱导恢复培养10d,分化培养每4week继代一次,分化培养8week(12h光周期,25℃,2 000lx)。试验侵染2 600个宁麦13愈伤组织,获得127株再生植株,经分子检测,其中9株证实为阳性转化植株。
译  名：
Agrobacterium-mediated Transformation by Mature Embryo of Wheat Ningmai 13
作  者：
JIANG Su,MA Hong-xiang,ZHANG Xu,YU Gui-hong,SUN Xiao-bo(Jiangsu Provincial Key Lab of Agribiology,Jiangsu Academy of Agricultural Science,Nanjing,Jiangsu 210014,China)
关键词：
Wheat(Triticum aestivum);Ningmai 13;Mature embryo;Agrobacterium tumefaciens;genetic transformation
摘  要：
Ningmai 13 is one of the main cultivated soft wheat varieties in Yangtze River area.To establish an effective system of genetic transformation in Ningmai 13 is useful for the further improvement of aimed characters in this variety.Wheat calli derived from mature embryos of Ningmai 13 were used as recipients to transfer plasmid pCXK1301 or plasmid pAtMYB12u by using the Agrobacterium strain GV3101.Preculture duration,co-culture duration and desiccation treated were optimized to set up the transformation system.The genetic transformation mediated by Agrobacterium tumefaciens for Ningmai 13 mature embryo was as follows: 6 d precultured mature embryo calli infected by Agrobacterium strain GV3101 containing plasmid pAtMYB12u,the concentration of Agrobacterium was OD600=0.6 and the infection time was 60 min,the co-culture condition was desiccation treated and the co-culture time was 4-5 d(23℃,dark culture).Afterwards,the infected recipient was inducted and recovered for 10 d,and regenerated for 8 weeks with sub-culturing every 4 weeks(12 h photoperiod,25℃,2 000 lx).2 600 mature embryo calli of Ningmai 13 were used for transformation and 127 regenerated plants were obtained,9 of them were transgenic plants validated by PCR.

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