蝴蝶兰花青素苷合成途径关键酶基因的克隆与表达

摘要

Four key enzyme genes (CHS ,DFR ,F3′5′H ,A NS) involved in anthocyanins synthesis were cloned from the flower petals of Phalaenopsis with homology sequence cloning .The length of the sequence was 302 bp ,275 bp ,285 bp and 285 bp , respectively . Sequence analysis showed that these protein shared high similarity to other proteins from Dendrobium ,Cymbidium et al ,ranging from 81% -95% ,67% -91% ,68% - 92% and 83% -89% ,respectively .The results of phylogenetic analysis were in agreement with those described in plant taxonomy .Results of Real-time fluorescence quantitative PCR analysis showed that the relative expression level of CHS and F3′5′H gene was the highest in the flower large bud and early flowering stages ,and then decreased in blooming .The expression level of CHS and F3′5′H gene in petals and lip was greater than in sepals ,the trace expression revealed in Leaves and roots .%采用 RT-PCR 技术从蝴蝶兰‘满天红’花瓣中克隆获得花青素苷生物合成途径中 CH S 、 DFR 、 F3′5′H和 A NS 等4个关键酶基因的保守序列，长度分别为302、275、285和285 bp 。序列分析表明：4个关键酶蛋白与石斛兰、大花蕙兰等其他植物来源的花青素苷生物合成相关蛋白均具有较高的同源性，分别为81％～95％、67％～91％、68％～92％和83％～89％。系统进化分析表明4个关键酶基因的系统进化基本上符合植物分类学分类。实时荧光定量 PCR 检测结果表明， CH S 和 F3′5′H 基因在大花蕾期及始花期的表达量最高，盛花期表达量降低；在花瓣、唇瓣中的表达量大于萼片，在叶片、根中仅有微量表达。