百脉根ACC氧化酶基因LcACO1的克隆及序列分析  

Cloning and Sequence Analysis of 1-Aminocyclopropane-1-Carboxylic Acid Oxidase Gene LcACO1 from Lotus corniculatus

作　　者：马兰[1] Ma Lan(Xi＇an University, School of Biological and Environmental Engineering, Xi＇an, 710100)

机构地区：[1]西安文理学院,生物与环境工程学院,西安710065

出　　处：《分子植物育种》2017年第2期447-453,共7页Molecular Plant Breeding

基　　金：西安市科技计划项目(CXY1443WL23);陕西省教育厅科研项目(2015DC031)共同资助

摘　　要：植物激素乙烯参与植物生长发育多个生理过程的调控。ACC氧化酶是植物乙烯合成途径中的关键合成酶。本研究依据其它物种ACC氧化酶序列信息设计简并引物,并结合RT-PCR和快速扩增c DNA末端技术从牧草百脉根中首次克隆到ACC氧化酶基因的全长c DNA序列,命名为LcACO1。LcACO1全长c DNA为1 012 bp,具有一个924 bp ORF可编码307个氨基酸残基。生物信息学分析表明,LcACO1是含有Fe2+依赖的加氧酶结构域的稳定亲水蛋白,没有信号肽和跨膜结构域,亚细胞定位于细胞质。与其它植物中的ACO蛋白进行同源性比对显示,LcACO1蛋白与鹰嘴豆中ACO蛋白的一致性高达90%。实时荧光定量PCR表明,LcACO1基因在百脉根不同组织器官中差异表达,根中表达量最高,推测其主要在根的生长发育调节方面发挥作用。Ethylene, one plant phytohormone, plays important roles in the various aspects of plant growth and development. ACC oxidase（ACO） is the key enzyme in the ethylene biosynthetic pathway in high plant. According to the conserved sequences of ACO from other plants, degenerate primers were designed and the full-length c DNA of a novel ACO gene from Lotus corniculatus were first obtained with RT-PCR and rapid amplification of c DNA ends（RACE）, which was named as LcACO1. LcACO1 is 1 012 bp in length with a 924 bp ORF, which encodes a predicted polypeptide of 307 amino acids. Bioinformatics analysis showed that LcACO1 without signal peptide and transmembrane domain was a stable hydrophilic protein and located in the cytoplasm, and contains a Fe（II）-dependent oxygenase domain. The results of homology analysis showed that LcACO1 protein shared a 90%identities with Cicer arietinum. Quantitative RT-PCR analysis revealed that LcACO1 expressed variously in different organs and the expression level in root was the highest, which indicated that LcACO1 maybe play roles on root development in L.corniculatus.

关 键 词：百脉根 ACC氧化酶 基因克隆 生物信息学分析 表达模式 

分 类 号：S541.6[农业科学—作物学] Q943.2[生物学—植物学]