红花2个G6PDH基因的克隆与表达特征分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-1123

摘要/Abstract

摘要：

目的葡萄糖-6-磷酸脱氢酶（G6PDH）在植物生长发育及逆境响应中起重要作用，旨在克隆红花（Carthamus tinctorius L.）G6PDH基因，解析其序列与表达特征，并探究其在非生物胁迫响应中的作用，为红花抗逆育种提供候选基因。方法基于红花基因组数据鉴定并克隆G6PDH基因，进行生物信息学分析；利用RT-qPCR技术分析其组织表达模式及对激素（MeJA、ABA、SA）和干旱、盐、碱胁迫的响应；同时，采用紫外分光光度法测定非生物胁迫下G6PDH的酶活性变化。结果从红花中鉴定并克隆了2个G6PDH家族成员（CtG6PDH1与CtG6PDH2），分别定位于1号和3号染色体上。亚细胞定位和系统进化分析表明，CtG6PDH1为胞质型，定位于细胞质；CtG6PDH2为质体P1型，定位于叶绿体。启动子区域含有光响应、植物激素响应和胁迫响应相关顺式作用元件。组织表达显示，CtG6PDH1在根中表达较高，CtG6PDH2在叶片中表达量最高。干旱及盐碱胁迫下，CtG6PDH1显著上调表达，而CtG6PDH2的表达受多种激素诱导。酶活性检测进一步证实，干旱与盐碱胁迫后胞质型G6PDH的活性显著升高。结论在红花中鉴定并克隆到2个G6PDHs基因家族成员，分别定位于细胞质和叶绿体。其启动子区域含有参与生长发育、激素响应及胁迫响应的调控元件。胞质型CtG6PDH1在红花响应干旱、盐、碱等非生物胁迫过程中发挥关键作用。

关键词: 葡萄糖-6-磷酸脱氢酶, 克隆, 亚细胞定位, 表达分析, G6PDH酶活性, 红花

Abstract:

Objective Glucose-6-phosphate dehydrogenase (G6PDH) plays a crucial role in plant growth, development, and stress responses. This study aimed to clone the G6PDH genes from safflower (Carthamus tinctorius L.), analyze their sequences and expression patterns, and investigate their involvement in abiotic stress responses. These findings will provide valuable candidate genes for stress-resistant safflower breeding. Method Based on the genome data of safflower, G6PDH genes were identified and cloned, followed by bioinformatic analysis. Using RT-qPCR, the tissue-specific expression patterns of CtG6PDH genes and their responses to hormones (MeJA, ABA, and SA) and drought, salt and alkali stress were analyzed. Additionally, the activities of glucose-6-phosphate dehydrogenase (G6PDH) under these abiotic stresses were determined by ultraviolet spectrophotometry. Result Two G6PDH family members (CtG6PDH1 and CtG6PDH2) were identified and cloned from the safflower genome, located on chromosomes 1 and 3, respectively. Subcellular localization and phylogenetic analysis revealed that CtG6PDH1 is a cytosolic isoform localized in the cytoplasm, while CtG6PDH2 belongs to the plastidial P1-type and is located in chloroplasts. Their promoter regions contain multiple cis-acting elements associated with light response, phytohormone signaling, and stress responses. Tissue expression analysis showed that CtG6PDH1 exhibits higher expression in roots, while CtG6PDH2 demonstrated the highest expression in leaves. Under drought and saline-alkaline stress, the cytosolic CtG6PDH1 gene was markedly upregulated, while CtG6PDH2 expression exhibited induction by multiple phytohormones. Further enzymatic assays confirmed that cytosolic G6PDH activity was significantly enhanced under both drought and saline-alkali stress conditions. Conclusion Two G6PDH genes were identified and cloned from safflower, exhibiting distinct subcellular localizations (cytosol and chloroplast). Promoter analysis revealed putative cis-regulatory elements associated with growth, hormone responses, and stress adaptation. RT-qPCR and enzyme activity analyses demonstrated that the cytosolic CtG6PDH1 plays a major role in safflower’s response to abiotic stresses, including drought, salt, and alkali.

Key words: glucose-6-phosphate dehydrogenase (G6PDH), clone, subcellular localization, expression analysis, G6PDH activity, safflower

秦娟, 卢蔚雯, 李佳徽, 李晨, 李斌, 王治业. 红花2个G6PDH基因的克隆与表达特征分析[J]. 生物技术通报, 2026, 42(8): 217-228.

QIN Juan, LU Wei-Wen, LI Jia-Hui, LI Chen, LI Bin, WANG Zhi-ye. Cloning and Expression Analysis of Two G6PDH Genes from Safflower[J]. Biotechnology Bulletin, 2026, 42(8): 217-228.

图/表 9

图1 红花G6PDHs基因的克隆与染色体定位

Fig. 1 Cloning and chromosome localization of G6PDHs gene in safflower (Carthamus tinctorius L.)

图2 CtG6PDH蛋白磷酸化位点、二级结构及三级结构的预测

Fig. 2 Prediction of the phosphorylated site, secondary structure and tertiary structures of CtG6PDH protein

图3 不同植物G6PDH蛋白的系统进化树

Fig. 3 Phylogenetic tree of G6PDH proteins in different plants

图4 CtG6PDH保守结构域及多序列比对分析

Fig. 4 Analysis of conserved domains and multiple sequence alignment of CtG6PDH

图5 CtG6PDHs的启动子顺式作用元件分析

Fig. 5 Analysis of promoter cis-acting elements of the CtG6PDHs gene

图6 CtG6PDH1与CtG6PDH2的亚细胞定位

Fig. 6 Subcellular localization of CtG6PDH1 and CtG6PDH2

图7 CtG6PDHs基因在不同组织中的表达模式

Fig. 7 Expression patterns of CtG6PDHs genes in different tissues

图8 CtG6PDHs在不同胁迫处理及激素诱导下的表达模式不同小写字母表示在P<0.05水平上差异显著

Fig. 8 CtG6PDHs expression patterns under different stress treatments and hormone inductionDifferent lowercase letters indicate the significant difference at P<0.05 level

图9 不同胁迫处理下G6PDH酶活性测定* P<0.05， ** P<0.01

Fig. 9 Determination of G6PDH enzyme activity under different stress treatments

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