辣椒泛素结合酶变体类似蛋白基因克隆及其与氮吸收利用的相关性分析  

Cloning of E2 Variant Like Protein Gene in Pepper and Its Correlation With Nitrogen Absorption and Utilization

作　　者：王春萍[1] 张世才[2] 李怡斐[2] 杨小苗 段敏杰 黄启中[2] 黄任中[2] 吴红[1] WANG Chunping;ZHANG Shicai;LI Yifei;YANG Xiaomiao;DUAN Minjie;HUANG Qizhong;HUANG Renzhong;WU Hong(Chongqing Key Laboratory of Adversity Agriculture,Biotechnology Research Institute,Chongqing Academy of Agricultural Sciences,Chongqing 401329;Vegetable and Flower Research Institute,Chongqing Academy of Agricultural Sciences,Chongqing 401329)

机构地区：[1]重庆市农业科学院生物技术研究所逆境农业研究重庆市市级重点实验室,重庆401329 [2]重庆市农业科学院蔬菜花卉研究所,重庆401329

出　　处：《核农学报》2023年第3期442-448,共7页Journal of Nuclear Agricultural Sciences

基　　金：现代农业产业技术体系专项资金(CARS-24-A-04);重庆市技术创新与应用发展专项项目(cstc2020jscx-msxmX0054);川渝联合实施重点研发项目(cstc2021jscx-cylhX0002)。

摘　　要：UEVs蛋白是泛素结合酶E2的变体,可通过与E2形成复合物催化底物泛素化。为探究UEVs蛋白在氮吸收利用中的作用,本研究参考辣椒低氮胁迫相关转录组数据,利用反转录PCR方法从辣椒叶片中克隆了泛素结合酶变体类似蛋白基因,命名为CaUev1D-L,对其进行生物信息和系统进化分析,并对转CaUev1D-L基因烟草进行低氮胁迫后的表型和生理特性分析。结果表明,CaUev1D-L开放阅读框长402 bp,编码133个氨基酸,具有E2结合酶UBC结构域,比辣椒、烟草、番茄和马铃薯等茄科作物的Uev1D蛋白氨基酸序列末端少14个氨基酸,与Uev1D蛋白的UBC结构域存在6个氨基酸的差异,与辣椒等作物的Uev1D蛋白不在同一进化分支。将CaUev1D-L转入烟草,在正常氮素水平下,转基因植株的茎叶干重显著小于野生型植株;在低氮水平下,转基因烟草植株的茎叶干重和根干重均显著大于野生型植株。本研究结果为氮吸收利用分子机制研究和相关基因的发掘利用提供了参考。UEVs protein is a variant of ubiquitin binding enzyme E2,which can catalyze substrate ubiquitination by forming a complex with E2.In order to explore the role of UEVs protein in nitrogen absorption and utilization,the gene of ubiquitin binding enzyme variant like protein named CaUev1D-L,was cloned from pepper leaves by reverse transcription PCR based on the transcriptome data related to low nitrogen stress in pepper.The biological information and phylogenetic analysis were carried out,and the phenotype and physiology of transgenic tobacco after low nitrogen stress were analyzed.The results showed that CaUev1D-L has an open reading frame of 402 bp,encodes 133 amino acids and has the UBC domain of E2 conjugating enzyme.The amino acid sequence of Uev1D protein is 14 amino acids less than that of Solanaceae crops such as pepper,tobacco,tomato and potato,and there are 6 amino acids differential with the UBC domain of Uev1D protein.It is not in the same evolutionary branch as Uev1D protein of pepper and other crops.When CaUev1D-L was transferred into tobacco,the dry weight of transgenic plants shoot was significantly lower than that of wild-type plants under normal nitrogen level;at low nitrogen level,the dry weight of shoot and root of transgenic tobacco plants were significantly greater than those of wild-type plants.This study can provide a reference for the study of molecular mechanism of nitrogen absorption and utilization and the exploration and utilization of related genes.

关 键 词：辣椒 泛素化结合酶 UEVs 氮代谢 

分 类 号：Q943.2[生物学—植物学]