Cloning and preliminary functional analysis of the root gene HgAKR6C of Halogeton glomeratus

doi:10.11686/cyxb2023076

Abstract

Abstract:

Aldo-keto reductase （AKR） is a conserved core domain that constitutes Shaker family K⁺ channel protein and plays a key role in plant response to abiotic stress. In this research， Halogeton glomeratus， a halophyte found in the northwest of China， was studied. Based on the results of initial transcriptomic investigation， the salt tolerance gene HgAKR6C was selected and cloned. The total length of the coding sequence （CDS） region of HgAKR6C gene was 951 bp， encoding 317 amino acids. Phylogenetic tree analysis showed that HgAKR6C is closely related to AtAKR6C1 gene in Arabidopsis thaliana. An evaluation of subcellular localization suggests that the gene is primarily localized in the cytoplasm and nucleus. A qRT-PCR analysis of the root gene HgAKR6C showed that the peak expression was reached at 24 h of salt treatment. Construction of yeast heterologous expression vectors to transform defective strains revealed that the HgAKR6C gene may be involved in Na⁺ efflux and mediates K⁺ uptake. In summary， HgAKR6C has the function of regulating salt tolerance in H.glomeratus. The regulatory mechanism of the salt tolerance gene HgAKR6C in the roots of H. glomeratus needs to be further investigated and verified.

Key words: Halogeton glomeratus, HgAKR6C, gene cloning, salt tolerance regulatory mechanism

Shang-qin HU, Jun-cheng WANG, Li-rong YAO, Er-jing SI, Xiao-le MA, Ke YANG, Hong ZHANG, Ya-xiong MENG, Hua-jun WANG, Bao-chun LI. Cloning and preliminary functional analysis of the root gene HgAKR6C of Halogeton glomeratus[J]. Acta Prataculturae Sinica, 2024, 33(1): 61-74.

Figures/Tables 14

References 44

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引物名称 Name of primers	引物序列 Sequences of primers （5′→3′）
HgAKR6C-F1	GCTCTAGAATGCAGTACAAGAACCTAGG
HgAKR6C-R1	TCCCCGGGGAACAACGGCCTCAATTTTC
HgAKR6C-F2	CCGGAATTCATGCAGTACAAGAACCTAGG
HgAKR6C-R2	GCTCTAGAAACAACGGCCTCAATTTTC
HgAKR6C-q1F	AGGAAGCACATCGTTGAGGG
HgAKR6C-q1R	TTCATTGCCCGGACAGTCTC
HgActin-F	TGTTCTCAGTGGTGGTACAA
HgActin-R	GTGCCACCACCTTAATCTTC

引物名称 Name of primers	引物序列 Sequences of primers （5′→3′）
HgAKR6C-F1	GCTCTAGAATGCAGTACAAGAACCTAGG
HgAKR6C-R1	TCCCCGGGGAACAACGGCCTCAATTTTC
HgAKR6C-F2	CCGGAATTCATGCAGTACAAGAACCTAGG
HgAKR6C-R2	GCTCTAGAAACAACGGCCTCAATTTTC
HgAKR6C-q1F	AGGAAGCACATCGTTGAGGG
HgAKR6C-q1R	TTCATTGCCCGGACAGTCTC
HgActin-F	TGTTCTCAGTGGTGGTACAA
HgActin-R	GTGCCACCACCTTAATCTTC

理化性质Physical and chemical properties	预测结果Prediction results
编码的氨基酸数Number of amino acids	317
理论等电点Theoretical pI	6.25
蛋白质分子量Molecular weight （Da）	35151.21
分子式Formula	C₁₅₇₅H₂₄₇₃N₄₁₉O₄₆₈S₁₂
负电荷的残基总数（天冬氨酸+谷氨酸）Total number of negatively charged residues （Asparticacid+glutamicacid， Asp+Glu）	36
正电荷的残基总数（精氨酸+赖氨酸）Total number of positively charged residues （Arginine+lysine， Arg+Lys）	34
脂肪系数Aliphatic index	90.41
亲水性平均值Grand average of hydropathicity	-0.219
不稳定系数The instability index （II）	32.06

理化性质Physical and chemical properties	预测结果Prediction results
编码的氨基酸数Number of amino acids	317
理论等电点Theoretical pI	6.25
蛋白质分子量Molecular weight （Da）	35151.21
分子式Formula	C₁₅₇₅H₂₄₇₃N₄₁₉O₄₆₈S₁₂
负电荷的残基总数（天冬氨酸+谷氨酸）Total number of negatively charged residues （Asparticacid+glutamicacid， Asp+Glu）	36
正电荷的残基总数（精氨酸+赖氨酸）Total number of positively charged residues （Arginine+lysine， Arg+Lys）	34
脂肪系数Aliphatic index	90.41
亲水性平均值Grand average of hydropathicity	-0.219
不稳定系数The instability index （II）	32.06

定位Positioning	预测占比Forecast proportion （%）
细胞质Cytoplasm	60.9
细胞核Nucleus	13.0
线粒体Mitochondrium	13.0
细胞骨架Cytoskeleton	4.3