葡萄糖氧化酶基因转化棉花和抗性愈伤组织的获得

Obtained Callus Resistant to Kanamycin from Cotton Transformation by Agrobacterium tumefaciens with Go Gene

用葡萄糖氧化酶基因转化棉花 ,研究了影响转化和抗性愈伤组织获得的因素。结果表明 ,基因型和培养基构成是影响葡萄糖氧化酶基因转化棉花获得抗性愈伤组织的最关键因素 ,在本试验所采用的 8个基因型中 ,中棉所 1 3中 940 9(中棉所3 5)和中棉所 1 9较易诱导获得抗性愈伤组织 ,而珂字 2 0 1、中棉所 2 7等则较难 ;用于棉花遗传转化的感染时间以 7～ 1 0 min为宜 ,共培养时间以48h为宜 ;下胚轴比子叶易诱导获得抗性愈伤组织。在诱导愈伤组织的继代培养中 ,加入高浓度的卡那霉素是获得转化细胞的关键 ;

The paper studied the factors which influenced the cotton transformation by Agrobacterium tumerfaciens, and obtained callus resistance containing Glucose oxidase(Go) gene. The results indicated that the genotypes and medium were two key factors that influenced the cotton transformation. CCRI13, Zhong 9409 and CCRI19 were easier to obtain resistance callu while; Coker201 and CCRI27 were more difficult. The option of influenced time was 7~10 min, and the option of co culture time was 48 hours for cotton transformation by Agrobacterium tumefaciens. Hypocotyl was easier to obtain resistance callus than cotyledon. It was the key factor of adding high concentration kanamycin for obtaining transformed callus. Liquid medium was easier to obtaine resistance callus than solid medium.