根癌农杆菌介导的外源基因转化植物萌动种胚的研究

STUDIES ON TRANSFORMATION OF SEED-GERMINATING EMBRYOES OF PLANTS BY AGROBACTERIUM-MEDIATED FOREIGN GENES

本文建立的新型根癌农杆菌转化程序是以微伤处理的黄瓜、番茄受体态种胚为实验材料,探讨了萌动种胚作为农杆菌TiT-DNA转化受体的可能性;确定了转化体系中筛选条件及受体态种胚的形态学特征和生理学指标;短期内获得了转基因植株,并从生理生化及分子水平上证实了外源基因的转移、整合与表达。本文为农杆菌Ti质粒载体介导的外源基因向植物中的转移提供了一个简便易行的受体系统。

A new procedure for the Agrobacterium-mediated transformation was established using the seed-germinating mature embryoes of both Cucumis sativus and Lycopersicum esculentum. The susceptible seedgerminating embryoes were slightly wounded before they were cocultivated with the strain C 58 Cl Rif^r Agrobacterium tumefaciens containing the pGV 3850::neo 1103 Ti plasmid. The selective conditions for transformed as well as the morphological characteristics and physiological indexes of seed embryoes in recipient state were ascertained according to the effect of different kanamycin concentrations upon them. Nopaline synthase and neomycin phosphotransfcrase Ⅱ activities were assayed in the transgenic plants obtained within shortterm. It was also confirmed that the foreign genes had been integrated into the genome of transformed plants utilizing southern blot method. This paper offered a advantageous recipient system for transfer of foreign genes into plant mediated by Agrobacterium Ti plasmid.