Identification of G6PDH Gene Family in Chenopodium quinoa and Functional Analysis of CqG6PDH1 in Salt Tolerance

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

Abstract

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

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.

Figures/Tables 8

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

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

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

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

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

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

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

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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