摘要： 为探讨甜瓜的遗传转化技术,以甜瓜品种河套蜜瓜腋芽生长点做外植体,利用庆大霉素基因作为筛选基因,对影响农杆菌转化的菌液浓度、转化时间、共培养时间进行了正交试验。通过优化转化条件,建立了农杆菌介导的甜瓜腋芽生长点遗传转化体系。结果表明,采用培养20 d的无菌苗,以腋芽作为外植体,预培养1 d;用OD₆₀₀=0.7的菌液转化10 min后,黑暗中共培养2 d,然后用40 mg/L庆大霉素进行筛选;移栽后在叶片上喷洒20 mg/L庆大霉素筛选转基因植株,效果最好。通过对庆大霉素抗性苗进行PCR检测,初步验证外源基因已经整合进甜瓜基因组,转化率达到76%,幼苗移栽成活率为76.3%。

关键词: 甜瓜, 植物生长点, 遗传转化, 农杆菌

Abstract: A high-frequency transformation system has been established by Agrobacterium tumefaciens transfor- mation technology and using meristems of axillary buds as transformation receptor. External factors that affect the ef- ficiency of genetic transformation on the base of gentamicin were studied and optimized, such as the bacterial con- centration and the times of transformation and co-cultivation. The major results were as follows: seedling was grown for 20 days, then axillary buds excised and stabbed were infected with Agrobacterium tumefaciens which OD600 was 0. 7 for 10 min. Then after co-cultivation for 2 days, transformed plants were selected by 20 mg/L and 40 mg/L gen- tamicin on the leaves and rooting respectively. It was originally proved by the PCR that expression cassettes had been transferred into the genome. The positive transformation rate was 76% by the analysis of PCR. And the living rate of the transplanted transgenic plants was 76. 3% .

Key words: Melon, Plant apical meristem, Genetic transformation, Agrobacterium tumefaciens

中图分类号: 

• S652