《植物生理学报》 2021, 57(6): 1261-1270

橡胶草ERF3基因的克隆、亚细胞定位及表达分析

张云川，曹天澳，雷骥良，闫洁^*

石河子大学生命科学学院, 新疆石河子832000

通信作者：闫洁；E-mail: 1653842328@qq.com

摘　要：

AP2/ERF是植物中广泛存在的一类特有转录因子, 在植物的次生代谢产物生物合成中发挥着重要 的调控作用。本研究采用PCR技术从橡胶草中克隆得到一个AP2/ERF亚家族基因TkERF3, 用生物信息学 方法分析了TkERF3的结构、性质和进化关系; 利用实时荧光定量PCR检测该基因在不同组织和植物生 长调节物质乙烯利、茉莉酸处理下的表达情况; 同时运用洋葱瞬时表达技术对该基因进行亚细胞定位分 析。结果显示, TkERF3基因全长CDS序列为651 bp, 编码216个氨基酸, 蛋白质分子质量为25.14 kDa, 理论 等电点(pI) 5.46。系统进化分析表明TkERF3蛋白与朝鲜蓟ERF蛋白亲缘关系最近。亚细胞定位显示 TkERF3蛋白位于细胞核中。qRT-PCR分析发现TkERF3基因主要在根中表达; 并且在植物生长调节物质 乙烯利、茉莉酸处理下均表现为显著上调表达, 推测该基因可能参与到乙烯和茉莉酸激素介导调控的天 然橡胶合成信号转导途径。

关键词：橡胶草; TkERF3; 生物信息学分析; 亚细胞定位; 表达分析

资助：国家自然科学基金(31860070)

Cloning, subcellular localization and expression analysis of TkERF3 in  Taraxacum kok-saghyz

ZHANG Yunchuan, CAO Tian’ao, LEI Jilian, YAN Jie^*

College of Life Sciences, Shihezi University, Shihezi, Xinjiang 832000, China

Corresponding author: YAN Jie; E-mail: 1653842328@qq.com

Abstract:

AP2/ERF is a unique type of transcription factor that exists widely in plants, and it plays an important regulatory role in the biosynthesis of plant secondary metabolites. In this study, an AP2/ERF subfamily gene TkERF3 was cloned from Taraxacum kok-saghyz by PCR technology, and the structure, properties and evolutionary relationship of TkERF3 were analyzed by bioinformatics methods; real-time  fluorescent quantitative PCR was used to detect the gene expression in different tissues and plant growth  regulator ethephon and jasmonic acid treatment; at the same time, the onion transient expression technology was used to analyze the subcellular location of the gene. The results showed that the TkERF3 gene is  651 bp in length, encoding 217 amino acids, the molecular weight of the protein is 25.14 kDa, and the theoretical isoelectric point (pI) is 5.15. Phylogenetic analysis showed that TkERF3 protein is closely related  to artichoke ERF protein. The subcellular localization showed that TkERF3 protein was located in the nucleus. RT-qPCR analysis found that the expression level of TkERF3 gene in roots was higher than that in  other tissues; The TkERF3 gene was significantly up-regulated under the treatment of ethephon and jasmonic acid, indicating that this gene may be involved in the signal transduction pathway of natural rubber  synthesis mediated by ethylene and jasmonic acid hormone.

Key words: Taraxacum kok-saghyz; TkERF3; bioinformatics analysis; subcellular localization; expression analysis