影响根癌农杆菌介导水稻转化的因素分析-植物研究.pdf

21 　3 　　　　　　　　　　　　　　　　　　　　　　 2001　7 Vol.21　No.3　　　　　　　　　　　BULLETIN OF BOTANICAL RESEARCH July, 　2001 尹鸿瑛　安韩冰　安利佳 (, 　116012) 　　根癌农杆菌与来自水稻成熟种子盾片的愈伤组织共培养, 将GUS 基因导入水稻愈伤组 织, 并获 了转基因植株。通过比较影响根癌农杆菌转化频率的各种因素, 表明激素配比为2, 4 -D 1mg/L 、TD 0.5mg/L、NAA 1mg/L 时, 可以大大促进籼稻愈伤组织的分化能力;酚类化合物 的加入使农杆菌的转化频率提高8.9%～23.5%;共培养时农杆菌的稀释方式及适当调整潮霉素 (hygB)的使用浓度影响到农杆菌的转化频率。 　水稻愈伤组织;植株再生;根癌农杆菌;转化频率 ANALYSISOFFACTORSAFFECTINGTHE EFFICIENCYOF AGROBACTERIUM -MEDIATED RICE TRANSFORMATION YIN Hong-ying 　AN Han-bing 　AN Li-jia (Bioenqineering Department, Dalian University of Technology , Dalian, 116012) Abstract　The rice calli were infected and transformed by A.tumefaciens strain LBA4404 harboring Ti plasmid pTOK233.After continuous hygB selection, stable hygromycin-resistant, GUS-posi- tive calli and plants were recovered.Some putative transgenic R0 plants were conducted with molecu- lar assays, the results indicated that foreign genes had been stably integrated in the genome of trans- genic rice.The results suggested that the ingredients of holmen of 2, 4-D 1mg/L、TD 0.5mg/L、 NAA 1mg/L were efficient for Agrobacterium-mediated transformation of rice, besides, the addi- tion of AS increased the efficiency of transformation by 8.9%～23.5%.At the same time, we found that both the dilution of agrobacteria and the adjusting of the concentration of hygB affected the effi- ciency of transformation. Keywords　rice calli;transgenic rice;A .tumefaciens;frequency of transformation 　　1988 , [2] 。 , ( 、PEG )( , 。 )。, , , , 、 , , 。 。 1　 [1] , 1.1　、 :(1