藜麦G6PDH基因家族的鉴定及CqG6PDH1耐盐性分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-1388

摘要/Abstract

摘要：

目的基于藜麦基因组和转录组数据，鉴定藜麦G6PDH基因家族成员，并探究候选基因CqG6PDH1在耐盐方面的功能，为藜麦耐盐机制的研究提供理论依据和基因资源。方法根据拟南芥AtG6PDH和水稻OsG6PDH基因家族成员的蛋白序列和G6PDH对应的隐马尔可夫文件PF02781和PF00479，对CqG6PDH基因家族成员进行鉴定。利用TBtools、MEGA、MEME和WoLF PSORT等工具分析基因家族成员的理化性质、进化关系、基因结构、保守基序、染色体定位、亚细胞定位和表达模式。运用RT-qPCR分析CqG6PDH1在不同组织和盐胁迫下的表达模式，通过转化拟南芥对其耐盐性进行功能验证。结果鉴定获得6个CqG6PDH基因家族成员，编码蛋白的氨基酸数目为519-722，预测蛋白分子量为59.30-79.58 kD，理论等电点为5.24-8.69。CqG6PDHs在第4、5、11、12和14染色体上均有分布，预测其编码的蛋白定位于细胞质和叶绿体。RNA-Seq数据和RT-qPCR表明，CqG6PDH1响应盐胁迫。转基因结果显示，分别用175和250 mmol/L NaCl处理转基因拟南芥种子和幼苗，过表达CqG6PDH1拟南芥的萌发率和存活率均显著提高，失水速率和丙二醛含量降低，SOD和POD活性及细胞膜稳定性增强。结论从藜麦基因组中鉴定到6个CqG6PDH基因家族成员，其中，过表达CqG6PDH1增强转基因拟南芥的耐盐性，其在应答盐胁迫的调控过程中具有重要作用，是改良作物耐盐性的重要候选基因。

关键词: 藜麦, G6PDH基因家族, 生物信息学分析, CqG6PDH1, 耐盐性

Abstract:

Objective Based on the information of genome and transcriptome, the members of the G6PDH gene family in quinoa were identified through bioinformatics methods. The function of the candidate gene G6PDH1 in salt tolerance was investigated. This study provides theoretical basis and gene resources for researching the salinity tolerance mechanisms in quinoa (Chenopodium quinoa Willd.). Method Based on the protein sequences of AtG6PDHs in Arabidopsis thaliana and OsG6PDHs in Oryza sativa, and the reported hidden markov model (HMM) files PF02781 and PF00479, the CqG6PDH gene family members were identified. Various bioinformatics tools such as TBtools, MEGA, MEME and WoLF PSORT were used to analyze the protein physicochemical properties, evolutionary relationships, gene structures, conserved motifs, chromosomal distribution, subcellular localization, and expression pattern of CqG6PDH gene family members. Additionally, RT-qPCR was used to analyze the expression pattern of CqG6PDH1 in different tissues and under salt stress, and A. thaliana was used to transform for validating the function of CqG6PDH1 in salt tolerance. Result Six CqG6PDH gene family members were identified, encoding proteins with 519-722 amino acids, with predicted molecular weights ranging from 59.30 to 79.58 kD and theoretical isoelectric points ranging from 5.24 to 8.69. CqG6PDHs were distributed on chromosomes 4, 5, 11, 12, and 14, and it is predicted that their encoded proteins were localized in the cytoplasm and chloroplast. RNA-Seq data and RT-qPCR indicated that CqG6PDH1 responded to salt stress. The seeds and seedlings of transgenic Arabidopsis were treated with 175 and 250 mmol/L NaCl, respectively, the germination rate and the survival rate of Arabidopsis overexpressing CqG6PDH1 significantly increased, accompanied by low water loss rate and malondialdehyde (MDA) content, as well as enhanced SOD activity, POD activity and cell membrane stability. Conclusion Six CqG6PDH genes are identified in the quinoa and over-expressing CqG6PDH1 in Arabidopsis resulted in enhanced tolerance to salt. CqG6PDH1 is an important candidate gene for improving crop tolerance to salt and plays an important role in the regulatory process of responding to salt stress.

Key words: quinoa, G6PDH gene family, bioinformatics analysis, CqG6PDH1, salinity tolerance

王亦学, 刘佳慧, 许帆, 胡鑫, 王育川, 侯丽媛, 吴慎杰, 郝曜山, 董艳辉. 藜麦G6PDH基因家族的鉴定及CqG6PDH1耐盐性分析[J]. 生物技术通报, 2026, 42(10): 98-107.

WANG Yi-xue, LIU Jia-hui, XU Fan, HU Xin, WANG Yu-chuan, HOU Li-yuan, WU Shen-jie, HAO Yao-shan, DONG Yan-hui. Identification of G6PDH Gene Family in Chenopodium quinoa and Functional Analysis of CqG6PDH1 in Salt Tolerance[J]. Biotechnology Bulletin, 2026, 42(10): 98-107.

图/表 8

表1 藜麦G6PDH基因家族成员信息

Table 1 Information about G6PDH gene family members of quinoa (Chenopodium quinoa Willd.)

基因 Gene	基因ID Gene ID	染色体位置 Chromosome distribution	长度 Length （aa）	分子量 MW （kD）	等电点pI	不稳定系数 Instability index	总平均亲水性 GRAVY	亚细胞定位 Subcellular location
CqG6PDH1	AUR62018096-RA	Chr.04，36190960-36202858	519	59.42	6.71	42.68	-0.419	细胞质Cytoplasm
CqG6PDH2	AUR62006786-RA	Chr.05，76135361-76150678	722	79.58	7.24	43.99	-0.266	叶绿体Chloroplast
CqG6PDH3	AUR62031998-RA	Chr.11，28856918-28875861	578	65.58	5.24	43.16	-0.537	细胞质Cytoplasm
CqG6PDH4	AUR62000441-RA	Chr.12，4929849-4939869	652	72.92	6.18	47.43	-0.129	叶绿体Chloroplast
CqG6PDH5	AUR62005655-RA	Chr.14，55157363-55161143	569	64.48	8.04	43.28	-0.380	叶绿体Chloroplast
CqG6PDH6	AUR62029100-RA	Chr.00，39600961-39607326	527	59.30	8.69	46.13	-0.237	叶绿体Chloroplast

图1 藜麦、拟南芥和水稻G6PDH基因家族的系统发育进化树

Fig. 1 Phylogenetic tree of G6PDH gene family in Chenopodium quinoa, Arabidopsis thaliana and Oryza sativa

图2 藜麦G6PDH基因家族的基因结构（A）和保守基序（B）

Fig. 2 Gene structures (A) and conserved domains (B) of G6PDH gene family in C. quinoa

图3 藜麦G6PDH基因家族在不同组织（A）和盐胁迫（B）的表达热图

Fig. 3 Expression heat map of CqG6PDH gene familyin different tissues (A) and salt stress (B) in C. quinoa

图4 藜麦CqG6PDH1的克隆（A）及组织（B）和盐胁迫（C）表达分析* P<0.05， ** P<0.01. The same below

Fig. 4 Gene cloning of CqG6PDH1 in quinoa (A), and expression analysis in different tissues (B) and salt stress (C)

图5 CqG6PDH1与其他物种的氨基酸序列比对XP_057515678.1：苋菜；NP_001241264.1：大豆；NP_001330361.1：拟南芥；XP_011098824.1：芝麻；NP_001275397.1：马铃薯；XP_002534490.2：蓖麻

Fig. 5 Amino acid sequence alignment of CqG6PDH1 with other speciesXP_057515678.1: Amaranthus tricolor; NP_001241264.1: Glycine max; NP_001330361.1: Arabidopsis thaliana; XP_011098824.1: Sesamum indicum; NP_001275397.1: Solanum tuberosum; XP_002534490.2: Ricinus communis

图6 过表达CqG6PDH1拟南芥种子在NaCl处理下的表型分析A：萌发表型；B：CqG6PDH1的表达分析；C：萌发率统计

Fig. 6 Phenotype analysis of Arabidopsis seeds with over-expressing CqG6PDH1 under NaCl treatmentA: Germination phenotype. B: Expression analysis of CqG6PDH1. C: Statistics of germination rate

图7 过表达CqG6PDH1拟南芥幼苗在NaCl处理下的表型（A）及生理指标测定（B-G）

Fig. 7 Phenotypes (A) of Arabidopsis seedlings with over-expressing CqG6PDH1 under NaCl treatment and determination of physiological indexes (B-G)

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