甜瓜ACC氧化酶反义基因植物表达载体的构建及对烟草的转化

Construction of an antisense CM-ACO1 gene plant expression vector and its utilization in transformation of tobacco

应用反义基因技术抑制水果成熟过程中内源乙烯的合成,从而培育耐贮运品种是解决水果延熟保鲜难题的可行新方法。一个来自甜瓜的ACC氧化酶cDNA全序列被反向插入到植物表达载体pBI121中,从而构建了甜瓜ACC氧化酶反义基因植物表达载体。用该反义基因载体转化烟草获得转基因植株,并且转基因在转化植株的自交子代中发生孟德尔分离,由此验证了该载体构建的成功和可用性。此项研究为下一阶段用该反义基因载体转化甜瓜栽培品种做了充分必要准备。

Improving keeping quality of melon fruit is a difficult problem. Applying antisense gene strategy to reduce ethylene biosynthesis is a new feasible method to breeding melon cultivars which softening and senescence would be delayed. An ACC oxidase cDNA isolated from melon （Cucumis melo L.） fruit was constructed into a plant expression vector pBI121 in antisense orientation, and then transformed tobacco. The construction of the antisense gene vector is successful and usable, which was proved by the evidence that the antisense gene was transferred into tobacco and separated in self-pollinated progeny according to Mendel＇ s the law of segregation. These results gave a sufficient and essential preparation to transform melon main cultivars, for the purpose of increasing their keeping quality.