水仙胚性愈伤的获得及农杆菌介导GUS基因的遗传转化.pdf

33 4 ( ) ol. 33, No. 4 2009 7 Journal ofNanjing ForestryUniversity (NaturalScience Edition) July, 2009 G US 魏开发 (漳州师范学院生物科学与技术系, 福建 漳州 363000) : 用正交实验设计对前期影响水 胚性愈伤诱导和分化关联性大的因素进行综合分析, 结果显示,培养基类型激 素及处理方式不同添加物等对水 外植体胚性愈伤诱导及分化的影响是基于培养基的多因素协同效应水 适宜 胚性愈伤诱导培养基是:改良MB+ NAA(0. 2mg/L) + 6-BA( 2mg/L) + ABA (2mg/L)+ AgNO (5mg/L); 适宜的分化 3 培养基是:改良 N6+ 2, 4-D(0. 5mg/L) + 6-BA(2. 5mg/L) + NAA( 0. 2mg/L) + ABA( 2mg/L)实验中观察到水 体 细胞胚的发生, 在胚性愈伤诱导和分化体系优化基础上实现农杆菌介导 GUS 基因转化, 稳定表达抗性愈伤 GUS染色 显示最佳胚性愈伤诱导培养基能促进水 外植体感受态的形成,有利于外源基因的接受, PCR检测表明实验中获得 3 株转 GUS 基因阳性植株 : 水 ; 胚性愈伤; 农杆菌介导法; 遗传转化; G US 基因 : Q785 : A : 1000- 2006( 2009) 04- 0033- 05 Studies on som atic em bryogenesis of Chinese narcissu s and GUS gene genetic tran sform ation m ediated by A grobacterium tum efacien s WEIKai-fa ( Department of Biological Science and Technology, ZhangzhouN ormalUniversity, Zhangzhou 363000, China) Ab stract: Based on single-factor screening expermi ents ofembryogenic callus induction and differentiation ofChinese narcis- sus, comprehensive survey hasbeen carried outwith orthogonalexpermi entaldesign. The results showed thatembryonic callus induction and differentiation were under synergistic effects ofmutil-factors, including medium type, hormonal concentration and different supplements, the best embryogenic callus induction medium is: mi proved MB + NAA ( 0. 2 mg/L) + 6-BA (2mg/L) + ABA (2mg/L) + AgNO ( 5mg/L); the best differentiationmedium is: mi proved N6+ 2, 4-D( 0. 5mg/L) + 3 6-BA(2. 5mg/L) + NAA (