沙柳SpsDREB8基因的克隆与表达分析  

Cloning and Expression Analysis of SpsDREB8 Gene in Salix psammophila

作　　者：王雷[1] 张鑫 李安玉 薄高峰 郝璞 刘璐[1] 王文功 阿拉腾苏和 杨海峰[1] Wang Lei;Zhang Xin;Li Anyu;Bo Gaofeng;Hao Pu;Liu Lu;Wang Wengong;Alateng Suhe;Yang Haifeng(Forestry College of Inner Mongolia Agricultural University,Hohhot,010018)

机构地区：[1]内蒙古农业大学林学院,呼和浩特010018

出　　处：《分子植物育种》2022年第4期1145-1152,共8页Molecular Plant Breeding

基　　金：国家科技重大专项课题(2018ZX08020002-005-005);国家自然科学基金项目(31660216);内蒙古自治区应用技术研究与开发项目(2019GG004)共同资助。

摘　　要：脱水响应元件结合蛋白(DREB)是植物响应非生物胁迫应答的重要转录因子。本研究从沙柳(Salix psammophila)中克隆获得SpsDREB8基因,并对其进行AP2结构域多序列比对、构建进化树和预测蛋白结构等生物信息学分析,采用RT-qPCR方法对该基因在不同组织及逆境下的表达模式进行分析。结果表明,本研究成功克隆获得SpsDREB8基因,该基因全长864 bp,编码287个氨基酸,预测蛋白分子量为31615.84 Da,等电点为5.21,为亲水性蛋白,无信号肽区域,具有AP2保守结构域,属于AP2家族成员;基于系统进化树分析表明,该基因属于AP2/ERF转录因子家族DREB亚组A-2组,与杨柳科植物进化关系较近;实时荧光定量PCR结果显示,SpsDREB8基因在沙柳根、茎、叶、花中都有表达,其中在嫩茎中的表达量最高,根中的表达量最低;在高温、低温和NaCl处理下,SpsDREB8基因在1、2、4 h表达显著高于对照组;干旱处理24 h后,SpsDREB8基因表达显著上调。研究表明沙柳SpsDREB8可能在响应非生物胁迫过程中发挥重要作用。该研究结果为进一步探索沙柳抗逆分子机制提供了理论基础。Dehydration response element binding protein(DREB)is an important transcription factor for plants to respond to abiotic stress.In this study,we cloned the SpsDREB8 gene from Salix psammophila,and performed bioinformatics analysis such as AP2 domain multiple sequence alignment,phylogenetic tree construction and protein structure prediction and used RT-qPCR method to analyze the expression pattern of the gene in different tissues and under adversity.The results showed that this study successfully cloned the SpsDREB8 gene.The SpsDREB8 was 864 bp in length,encoding 287 amino acids.The putative protein Sps DREB8 was 31615.84 Da in molecular weight and with a theoretical isoelectric point of 5.21.It was a hydrophilic protein with no signal peptide.It has an AP2 conserved domain and belongs to the AP2 family.Based on phylogenetic tree analysis,this gene belongs to AP2/ERF transcription factor family DREB subgroup A-2 group,and has a close evolutionary relationship with Salicaceae plants.Real-time quantitative polymerase chain reaction results showed that SpsDREB8 could be detected in roots,stems,leaves and flowers of S.psammophila,among which the highest expression was in young stems and the lowest expression was in roots;under high temperature,low temperature and NaCl treatments,the expression of SpsDREB8 gene at 1,2 and 4 h was significantly higher than that of the control group;after 24 h of drought treatment,the expression of SpsDREB8 gene was significantly up-regulated.It shows that SpsDREB8 may play an important role in the process of S.psammophila coping with abiotic stress.The results of this study provided the foundation for further exploration of the molecular mechanism of S.psammophila.

关 键 词：沙柳(Salix psammophila) SpsDREB8 基因克隆 非生物胁迫 表达分析 

分 类 号：S793.9[农业科学—林木遗传育种]