绣球花花色相关基因HmF3H的克隆及其表达分析

花色是植物重要的观赏性状之一,而黄烷酮羟化酶是植物花色素物质积累的关键酶,对植物花色差异具有重要影响。本研究利用PCR技术从盛花期蓝色花绣球品种‘无尽夏’不孕花中克隆获得了Hm F3H基因的全长序列,该基因的ORF序列长度为1 095 bp,编码364个氨基酸,编码蛋白的理论分子量为41.08 k D,理论等电点为5.48,为亲水性蛋白,该蛋白属于PLNO2515超家族成员,具有2-酮戊二酸双加氧酶结构域。系统进化树分析表明,该蛋白与咖啡树和佛手瓜的亲缘关系最近。实时定量PCR分析表明,Hm F3H基因在绣球花不同组织器官中均有表达,红色绣球品种‘粉黛’盛花期不孕花比蓝色绣球品种‘无尽夏’盛花期不孕花中表达量高,说明Hm F3H基因对绣球花花色差异形成具有一定影响。本研究获得了绣球花Hm F3H基因序列及其表达情况,为探讨绣球花品种花色差异形成机制具有一定指导意义。

Cloning and Expression Analysis of HmF3H Gene Related to Flower Color of Hydrangea macrophylla

Flower color is one of the important ornamental traits of plants,and flavanone hydroxylase is a key enzyme in the accumulation of plant anthocyanidins,which has an important influence on plant color difference.In this study,the full-length sequence of Hm F3 H gene was cloned from the blue infertility flower of Hydrangea macrophylla variety’Endless Summer’by PCR.The ORF sequence length of this gene is 1095 bp,encoding 364 amino acids,the theoretical molecular weight of the encoded protein is 41.08 k D and the theoretical isoelectric point is 5.48.It is a hydrophilic protein belonging to the PLNO2515 superfamily and having a 2-ketoglutarate dioxygenase domain.Phylogenetic tree analysis showed that the protein was most closely related to Coffea arabica and Iochroma edule.Real-time quantitative PCR analysis showed that Hm F3 H gene was expressed in different tissues and organs of Hydrangea macrophylla,and the expression level of red flower Hydrangea macrophylla variety’Fendai’in infertile flowers at full-bloom stage was higher than that of blue flower variety’Endless Summer’.It is indicated that the Hm F3 H gene has a certain influence on the flower color of Hydrangea macrophylla.In this study,the Hm F3 H gene sequence and expression of Hydrangea macrophylla were obtained,which is of certain guiding significance for exploring the formation mechanism of flower color difference between Hydrangea macrophylla varieties.