观赏海棠McF3'H的克隆及不同品种间表达差异分析

摘要

为了探究Mc F3'H基因与花色苷之间的关系,以苹果属观赏海棠‘王族’叶片总RNA为模板,通过RACE扩增,获得类黄酮3'-羟化酶(Flavonoid 3'-hydroxylase,F3'H)基因序列,其基因编码区共1 536 bp,编码511个氨基酸,命名为McF3'H.McF3'H编码的氨基酸序列与其他物种的F3'H蛋白具有较高相似性.对3种不同叶色的观赏海棠品种‘火焰'、‘绚丽'和‘王族’叶片不同发育时期的McF3'H表达量、花青苷含量进行测定分析.结果表明McF3'H基因在3种不同叶色类型的观赏海棠品种叶片中均有表达,常色紫叶类‘王族’叶片McF3'H相对表达量明显高于新叶有色类‘绚丽'和绿色叶‘火焰’,McF3'H表达量与花色苷含量的变化规律较为一致.说明McF3 'H在苹果属观赏海棠叶片花青苷代谢及色泽形成过程中具有重要作用.%In order to study the relationship between McF3'H gene and anthocyanins, the total RNA from the leaves of Malus crabapple 'Royalty' was used as the template and the full gene sequence of F3'H (Flavonoid 3'-hydroxylase ) gene was cloned by rapid-amplification of cDNA end (RACE). The gene was named as McF3'H,containing an 1 536 bp open reading frame and encoding a protein of 511 amino acids. McF3 H protein was highly homologous to other species. In addition, McFZ H gene expression and anthocyanins content were detected in the leaves of 'Flame', 'Radiant' and 'Royalty' in different developmental stage. The results showed the expression level of McF3'H in'Royalty' is higher than in 'Flame' and 'Radiant'. The expression of McF3'H gene and anthocyanins content are almost consistent. Those results suggest that McF3'H may play an important role in crabapple anthocyanins metabolism and leaf color formation.