鹤望兰黄烷酮3-羟化酶基因SrF3H的克隆及表达分析

Cloning and Expression of Flavanone 3-hydroxylase from Strelitzia reginae Banks

类黄酮3-羟化酶是类黄酮生物合成途径中的关键酶,对花色的形成起重要作用。本研究克隆了鹤望兰类黄酮3-羟化酶基因SrF3H,该cDNA全长1 333 bp(GenBank收录号:KC412257),具有完整的开放阅读框(ORF),共1 122个碱基,编码374个氨基酸。与其他物种的氨基酸序列都有一定的同源性,其中与高杯花的F3H同源性最高,达到83%。系统进化树分析显示,鹤望兰SrF3H与百合蛋白亲缘关系较近。应用半定量PCR分析表明,SrF3H在蓝色花瓣中表达最高,而在鹤望兰不同花发育阶段差异不明显。研究结果不仅为植物育种提供了重要基因资源,也为剖析鹤望兰花色形成机理奠定基础。

Flavanone 3-hydroxylase was a key enzyme in flavoniod biosynthesis, which played an important role in flower color. In this research, the full-length cDNA sequence of F3H gene （SrF3H） was cloned from Strelitzia reginae Banks. The cDNA sequence was 1 333 bp and included an intact open reading frame of 1 122 bp, encoding a polypeptide of 374 amino acids. The amino acid was highly conserved compared with other F3H homologues and shared up to 83% homology with F3H from Nierembergia sp.. Phylogenetic analysis indicated that SrF3H was more related to F3H from Lilium hybrid division. The semi-quantitative PCR analysis indicated that SrF3H showed the highest transcript abundance in the blue petals, but there was little difference in flower development stages. This research not only supplied an important genetic resource for plant breeding, but also proved a basis for explo- ring the mechanism of flower color in Strelitzia reginae Banks.