黄瓜CsERF025L转录因子的克隆及表达分析  

Cloning and Expression Analysis of CsERF025L Transcription Factor in Cucumber

作　　者：肖雅茹 贾婷婷 罗丹 武喆[1] 李丽霞 XIAO Ya-ru;JIA Ting-ting;LUO Dan;WU Zhe;LI Li-xia(College of Horticulture,Shanxi Agricultural University,Taigu 030801)

机构地区：[1]山西农业大学园艺学院,太谷030801

出　　处：《生物技术通报》2024年第4期159-166,共8页Biotechnology Bulletin

基　　金：山西农业大学科技创新基金项目(2018YJ05)。

摘　　要：【目的】ERF转录因子是AP2/ERF基因家族的一个亚族,在植物生长发育和抵抗外界胁迫方面具有重要作用,克隆CsERF025-like(CsERF025L)并分析其表达模式,为黄瓜CsERF025L功能研究奠定基础。【方法】通过RT-PCR从黄瓜茎尖中克隆CsERF025L,对CsERF025L进行蛋白理化性质、亚细胞定位、互作蛋白等生物信息学分析,采用实时荧光定量PCR分析CsERF025L在黄瓜中的表达模式。【结果】CsERF025L编码序列为510 bp,编码169个氨基酸,具有一个AP2/ERF结构域,属于AP2/ERF家族的ERF亚家族。CsERF025L蛋白是一种不稳定的疏水性蛋白,位于细胞核,蛋白CsERF025L与P450基因家族成员、BEE3转录因子、DIR1蛋白和MATE家族成员存在互作。CsERF025L启动子含有光响应、激素响应、应激响应等元件。基因CsERF025L在黄瓜种子时期表达量最高,显著高于叶、茎和根。比较CsERF025L在不同叶序黄瓜中的表达,发现对生叶序黄瓜cop1茎尖中CsERF025L表达量均显著高于互生叶序黄瓜R1。【结论】CsERF025L是一个位于细胞核的ERF转录因子,可能在黄瓜对生叶序性状的形成与调控中具有潜在功能。【Objective】ERF transcription factor is a subfamily of AP2/ERF gene family,which plays a key role in plant growth and development and resistance to external stress.Cloning CsERF025-like(CsERF025L)and analyzing its expression pattern may lay a foundation for studying the function of CsERF025L.【Method】CsERF025L from cucumber(Cucumis sativus L.)stem tip was cloned by RT-PCR,and bioinformatics analysis of CsERF025L protein physicochemical properties,subcellular localization and interacting proteins was performed.Realtime fluorescence quantitative PCR was used to analyze the expression characteristics of CsERF025L in cucumber.【Result】The encoding sequence of CsERF025L was 510 bp,encoding 169 amino acids,with an AP2/ERF domain,belonging to the ERF subfamily of the AP2/ERF family.CsERF025L protein was an unstable hydrophobic protein located in the nucleus.CsERF025L protein interacts with members of P450 gene family,BEE3 transcription factor,DIR1 protein and MATE family members.CsERF025L promoter contained light response,hormone response,stress response and other elements.The expressions of CsERF025L in the cucumber seeds was the highest,which was significantly higher than that in leaves,stems and roots.The expression patterns of CsERF025L in the cucumber with different phyllotaxis types were compared,and the expressions of CsERF025L in the stem tip of opposite phyllotaxis cop1 were significantly higher than those of cucumber alternate phyllotaxis R1.【Conclusion】The findings of this study suggest that CsERF025L is a nuclear-localized ERF transcription factor,potentially involved in the formation and regulation of cucumber alternate phyllotaxis.

关 键 词：黄瓜 CsERF025L转录因子 生物信息学 表达分析 叶序 

分 类 号：Q943.2[生物学—植物学] S642.2[农业科学—蔬菜学]