山新杨谷胱甘肽转移酶基因的生物信息学与表达模式分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0430

摘要/Abstract

摘要：

目的分析山新杨谷胱甘肽转移酶（GST）基因的表达模式，为深入研究木本植物GST基因的功能提供基础。方法克隆山新杨GST基因并进行生物信息学分析，通过荧光定量PCR技术分析GST基因在植物激素及非生物胁迫下的表达模式。结果克隆了3个PdbGST基因家族成员，分别命名为PdbGST1、PdbGST2和PdbGST3，cDNA全长分别为678、660和690 bp，编码氨基酸分别为225、219和229个，形成的蛋白质均为稳定亲水酸性蛋白，且均定位于细胞质。同时，这3个蛋白质均具有谷胱甘肽转移酶的典型结构，且其启动子区域具有能够响应生物或非生物胁迫以及植物激素的元件。此外，这3个基因的表达不同程度地受外源植物激素或非生物胁迫诱导。结论 3个PdbGST基因均能够不同程度地响应非生物胁迫或外源植物激素的诱导，可能参与到杨树对逆境胁迫的响应过程中。

关键词: 谷胱甘肽转移酶, 生物信息学分析, 基因表达模式, 山新杨

Abstract:

Objective To provide a basis for further study of the function of glutathione transferase (GST) gene in woody plants, the expression pattern of GST genes from Populus davidiana × P. bolleana were analyzed. Method The GST genes were cloned and analyzed by bioinformatics of Populus davidiana × P. bolleana. The expression pattern of GST genes under phytohormone and abiotic stresses were analyzed by real-time PCR. Result Three GST gene members from Populus davidiana × P. bolleana were cloned. The lengths of these genes were 678, 660 and 690 bp, encoding amino acids of 225, 219 and 229 aa, respectively. Meanwhile, all three proteins have the typical structure of glutathione transferase, and their promoter regions have elements that can respond to biotic or abiotic stresses as well as plant hormones. In addition, the expressions of these three genes were differently induced by exogenous phytohormones or abiotic stresses. Conclusion All the three PdbGSTs may respond to abiotic stresses or the induction of exogenous phytohormone, indicating that they may play roles in response to the stresses in poplar.

Key words: glutathione transferase, bioinformatics analysis, gene expression pattern, Populus davidiana × P. bolleana

黄颖, 遇文婧, 刘雪峰, 刁桂萍. 山新杨谷胱甘肽转移酶基因的生物信息学与表达模式分析[J]. 生物技术通报, 2025, 41(2): 248-256.

HUANG Ying, YU Wen-jing, LIU Xue-feng, DIAO Gui-ping. Bioinformatics and Expression Pattern Analysis of Glutathione S-transferase in Populus davidiana × P. bolleana[J]. Biotechnology Bulletin, 2025, 41(2): 248-256.

图/表 8

表1 本研究所用引物

Table 1 Primers used in this study

基因名称 Gene name	正向引物序列 Forward primer （5′-3′）	反向引物序列 Reverse primer （5′-3′）	用途 Purpose
PdbGST1	ATGGCAGAGGAAGTGAAGGTCTTTAG	TCATTTATGTGTTTCTTTTCTAAGAC	基因克隆
PdbGST2	ATGGCAGGAATAAAACTACTAGATTC	TCATTCAATCCCAAGCTTCTTCCTG	基因克隆
PdbGST3	ATGGCAAACACAGAGCTGGTGAAGC	TCAAGTCCGAAACCAGTTATGAGG	基因克隆
PdbGST1-RT	ACCAGAAAGCCAATGCTCGT	GCCTCCCCACCGAAGAATTT	荧光定量PCR
PdbGST2-RT	GGTCTGGAATGACAGGTCTCCTTTGC	GGGCTTGTCTCCAAGCTCTCCTTC	荧光定量PCR
PdbGST3-RT	CCACTGATGCCTCAAGATCCTTACG	CCAGGTAGCCAATGTTCTCTCCTCC	荧光定量PCR
PdbEF1-α	TGGGTCGTGTTGAAACTGGTGT	GGCAGGATCGTCCTTGGAGTTC	荧光定量PCR
Pdbactin	GCTGAGAGATTCCGTTGCCCTG	GGCGGTGATCTCCTTGCTCATT	荧光定量PCR

表2 PdbGST蛋白的理化性质

Table 2 Characteristics of PdbGSTs protein

基因名称

Gene name

基因长度

Length/bp

氨基酸长度

Protein/aa

蛋白质相对分子质量

Mw/kD

等电点

Isoelectric point

不稳定系数

Instability index （II）

亲疏水性

GRAVY

亚细胞定位

Localization

PdbGST1

PdbGST2

PdbGST3

图1 PdbGST蛋白的多序列比对KAG6769001、KAG6793805、KAG6757347：毛白杨；XP_034919935、XP_034900369、XP_034914570：山杨；XP_011004483、XP_011030495：胡杨；ANO39980、ANO40011、ANO40009：亚东杨；KAH8506839、KAH8495610：美洲黑杨；KAJ6992085、KAJ7014641、KAJ6981522：银中杨；XP_002307924、XP_006370194、XP_006378334：毛果杨

Fig. 1 Multiple alignment of PdbGST proteinKAG6769001, KAG6793805, KAG6757347: Populus tomentosa; XP_034919935, XP_034900369, XP_034914570: P. alba; XP_011004483, XP_011030495: P. euphratica; ANO39980, ANO40011, ANO40009: P. yatungensis; KAH8506839, KAH8495610: P. deltoides; KAJ6992085, KAJ7014641, KAJ6981522: P. alba×P. berolinensis; XP_002307924, XP_006370194, XP_006378334: P. trichocarpa

图2 PdbGST蛋白的进化树分析红星标注PdbGST蛋白

Fig. 2 Phylogenetic tree analysis of PdbGST in poplarRed stars indicate PdbGST protein

图3 PdbGST基因的启动子序列分析

Fig. 3 Analysis of promoter sequences of PdbGST

图4 PdbGST基因的组织特异性表达分析不同小写字母表示同一基因不同组织差异显著分析（P<0.05）

Fig. 4 Tissues expression analysis of PdbGSTs geneDifferent lowercase letters indicate significant differences between a gene in different tissues （P<0.05）

图5 PdbGST基因响应不同植物激素诱导的表达模式分析A：水杨酸（SA）；B：脱落酸（ABA）；C：1-氨基环丙基-1-羧酸（ACC）；不同小写字母表示同一基因不同处理时间的差异显著分析（P<0.05）。下同

Fig. 5 Expression profiles of PdbGSTs under different phytohormones inductionA: SA treatment; B: ABA treatment; C: ACC treatment. Different lowercase letters indicate significant differences between a gene during the treatment （P<0.05）. The same below

图6 PdbGST基因响应非生物胁迫的表达模式分析A：PEG6000；B：甘露醇；C：NaCl

Fig. 6 Expression profiles of PdbGSTs under abiotic stressesA: PEG6000; B: mannitol; C: NaCl

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