山新杨谷胱甘肽S-转移酶PdbGST基因的克隆与胁迫表达分析  

Analysis of Cloning,Identification and Expression Pattern of Glutathione S-transferases Gene in Poplar

作　　者：平晓帆 遇文婧 黄颖 PING Xiaofan;YU Wenjing;HUANG Ying(Forestry Protection Institute of Heilongjiang Province,Heilongjiang Harbin 150081)

机构地区：[1]黑龙江省森林保护研究所,黑龙江哈尔滨150081

出　　处：《林业科技》2024年第3期33-40,共8页Forestry Science & Technology

基　　金：黑龙江省省属科研院所科研业务费项目(CZKYF2022-1-C048,LKSB2024-2,LKSB2023-4,2572020BA08);中央高校基本科研业务费专项资金项目(2572020BA08)。

摘　　要：在植物中,谷胱甘肽S-转移酶是一类多功能酶,能够在其响应非生物胁迫中发挥作用。为研究该基因在杨树中的功能,本研究克隆了山新杨谷胱甘肽S-转移酶基因Pdb GST及其启动子序列,对其进行生物信息学分析和表达特征分析。结果显示,Pdb GST基因的开放阅读框(ORF)全长666 bp,编码221个氨基酸,其形成的蛋白质相对分子质量为25.57 k Da,为稳定亲水性酸性蛋白,定位于细胞质中。系统进化分析表明,山新杨Pdb GST蛋白与山新杨中的XP_034926648蛋白的同源性最高。对克隆获取的2 000 bp启动子序列进行分析,结构显示该启动子序列中,含有多种响应非生物胁迫的顺式作用元件。荧光定量PCR分析显示Pdb GST基因在山新杨根部表达量最高,其次在山新杨叶部也有较高表达量,相反在山新杨顶芽中表达量最低。此外,对Pdb GST基因响应非生物胁迫分析显示,山新杨根部Pdb GST基因受诱导后即出现持续且大量的表达。In plant,glutathione S-transferase(GST) refers to a multifunctional enzymes that plays an important role in abiotic stress modulation pathways.To investigate the function of GST in poplar trees,a PdbGST gene and its 2 000 bp promoter squence were cloned and analyzed in Populus davidiana ×P.bolleana,and the expression pattern of it were also analyzed by RT-qPCR.The results showed that the open reading frame(ORF) length of PdbGST is 666 bp,which encodes 221 amino acids with the molecular weight is 25.57 kDa.The PdbGST is a stable hydrophilic acidic protein located in the cytoplasm.Systematic evolutionary analysis reveals that PdbGST in Populus davidiana × P.bolleana has a close relationship with XP_034926648 in P.alba.The analysis of cis-acting elements in promoter sequence of PdbGST showed that,this sequence contains multiple cis-acting elements related to plant response to abiotic stress.RT-qPCR showed that PdbGST had the highest expression level in the roots of Populus davidiana×P.bolleana,followed by a higher expression level in the leaves,and the lowest expression level in the top buds.In addition,analysis of the response of PdbGST in response to abiotic stress showed that PdbGST in the roots of Populus davidiana×P.bolleana exhibited sustained and extensive expression after induction.

关 键 词：山新杨 谷胱甘肽S-转移酶 非生物胁迫 表达模式 

分 类 号：S762.8[农业科学—森林保护学]