类黄酮3′,5′羟基化酶(Flavonoid-3′,5-′hydroxylase,F3′5′H)是花色苷代谢途径中的一个关键酶,能使花色素合成趋向于形成蓝色的飞燕草色素,从而使花色向蓝紫色偏移。本研究从蓝紫色矮牵牛(Petunia hybrida)花瓣中克隆了编码F3′5’H的蓝色基因Hf1,并通过PCR技术获得百合花特异表达启动子(PchsA),将百合PchsA与Hf1基因融合,构建了百合花特异表达启动子调控的Hf1基因植物表达载体,通过农杆菌介导的叶盘法转化粉红色矮牵牛。抗性筛选和PCR检测鉴定转基因植株,结果表明,成功地获得了转基因阳性植株。
Flavonoid-3′,5′-hydroxylase(F3′5′H) is the key enzyme in the biosynthesis of anthocyanins,which can divert the production of delphinidin pigments.The total RNA was isolated from the blue flower of Petunia hybrida.In the research the blue gene named as Hf1 encoding F3′5′H were isolated from the blue-violet petunia(P.hybrida).And the flower-specific promoter in lily named PchsA,was also cloned by PCR.The plant expression vector of Hf1 driven by the flower-specific promoter PchsA was constructed,which transformed pink petunia using Agrobacterium-mediated transformation of petunia leaf discs.Transgenic petunia has been obtained by hygromycin resistance screening and PCR detection.