【作者】 徐鹤林； 陆春贵； 余文贵； 杨荣昌； 曹晓凤； 朱玉贤； 冻章良；

【Author】 Xu Helin Lu Chungui Yu Wengui Yang Rongxhang Cao Xiaofeng Zhu Yuxian Chen Zhangliang( Insilitute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014; College of Life Sciences, Peking University, Beijing 100871)

【机构】 江苏省农业科学院蔬菜研究所； 北京大学生命科学学院；

【摘要】 ACC合成酶cDNA被反向插入载体PMON316,通过三亲交配,将表达质粒转移到农杆菌中,采用叶盘法转化番茄子叶.经卡那霉素筛选及分子杂交检测,表明,反义ACC合成酶基因已在mRNA水平表达.对转基因番茄果实的成熟生理指标测定结果表明,转基因番茄成熟受到严重抑制,乙烯的峰值为正常番茄的25%～30%,PG活性有所下降,果实在室温下可贮藏1个多月,品质和其他性状与正常番茄相近.更多还原

【Abstract】 1 - aminocyclopropane - 1 - caboxylate synthetase (ACC synthetase) is a key regulatory enzyme in the biosynthetic pathway of the plant hormone ethylene. A full length cDNA encoding this enzyme was cloned from tomato fruits. Two copies of this gene were lig-ated head - to - tail and inserted into the plant transformation vector PEACC18 in an anti-sense orientation. The antisense ACC gene with the 35s promoter was transferred into leaf discs of lycopersion esculentum cultivar FA7 and FA8 using Agrobacterium tumefaciens. These leaf discs were placed on MS medium containing kanamycin and carbenicillin for callus induction. They were transferred step by step to different differentiation media, and numerous plants regenerated eventually. Leaves of these transgenic tomato plants were used for assaying the gene expression. Results of Southern blot showed that the antisense ACC gene was stably integrated into the genome of transgenic plants. Physiological analyses showed that the expression of the antisense ACC gene inhibited fruit ripening significantly. The peak of enthylene production was 30% ~ 25% that of normal fruits, and the climacteric was delayed by a week with a mean storage life of 30 - 50 days. The shelf - life of the transgenic fruit was doubled compared with that of the parental line.更多还原