小兰屿蝴蝶兰YUCCA基因家族鉴定及在花朵中的表达分析

Identification of YUCCA Gene Family and Analysis of Its Expression in Flowers of Phalaenopsis equestris

生长素合成基因YUCCA在植物根、茎、叶的形成,花、果实、种子的发育等生命过程中起着重要作用。本试验针对小兰屿蝴蝶兰(Phalaenopsis equestris) YUCCA基因家族进行研究,鉴定得到8个PeYUCCAs家族成员。对其理化性质、系统进化树、保守结构域及保守基序、基因结构、蛋白三维结构等生物信息进行分析,结果表明:氨基酸长度为400~426 aa,蛋白质分子量为43.22~47.92 kD,等电点为6.43~9.47;GAGPSG(FAD-binding)、GCGNSG (NADPH-binding)为Pe YUCCAs的保守结构域;同时其保守motif为16条,9条为所有成员所共有,且其基因结构存在2~4个内含子;PeYUCCAs的蛋白三维结构主要由多个α-螺旋、β-折叠及无规则卷曲组成;在构建的系统进化树中,YUCCA序列分为四个进化枝,GroupⅠ进一步分为两个亚类群,且PeYUCCAs成员在每个进化枝中都有分布。此外,RT-qPCR试验结果表明:PeYUCCA3在花朵不同时期及不同部位均有特异性表达,其中大蕾花期、合蕊柱花部的表达量最高;而PeYUCCA1、PeYUCCA5仅在合蕊柱花部中有高表达。由此推测PeYUCCA1、PeYUCCA3、PeYUCCA5可能在花器官生长发育过程,尤其在雌雄蕊发育中发挥着重要作用。

The auxin synthesis gene YUCCA plays an irreplaceable role in plant life processes such as the formation of roots,stems,leaves,and the development of flowers,fruits,and seeds.In this experiment,the YUCCA gene family of Phalaenopsis equestris was studied,and eight members of the PeYUCCAs family were identified.The bioinformatics analysis of physicochemical properties,phylogenetic tree,conserved structural domains and conserved motifs,gene structures,three-dimensional structures of protein showed that the amino acid length was between 400 and 426 aa,the protein molecular weight ranged from 43.22 to 47.92 kD,and the isoelectric point was between 6.43 and 9.47;GAGPSG(FAD-binding)and GCGNSG(NADPH-binding)were the conserved structural domains of PeYUCCA family members.PeYUCCAs exhibited 16 conserved motifs,nine of which were shared across all members,with gene structures including 2 to 4 introns.The three-dimensional structures of PeYUCCAs'Peprotein were mainly composed of multiple alpha helices,beta folds,and random coils.In the phylogenetic tree,the YUCCAs sequence was divided into four clades,and GroupⅠwas further divided into two subgroups,and the PeYUCCAs members were distributed in each clade.RT-qPCR results showed that PeYUCCA3 was specifically expressed in different stages and different parts of flowers,with the highest expression observed at the mature bud stage and in the gynandrium.PeYUCCA1 and PeYUCCA5 were highly expressed in the Gynandrium.It wes speculated that PeYUCCA1,PeYUCCA3 and PeYUCCA5 may play an important role in the growth and development of flower organs,especially in pistil and stamen development.