Cloning of garlic hexokinase gene AsHXK2 and expression analysis of its involvement in rhizosphere growth-promoting bacteria alleviating drought stress

doi:10.3969/j.issn.1004-1524.2022.09.11

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (9): 1925-1934.DOI: 10.3969/j.issn.1004-1524.2022.09.11

• Horticultural Science • Previous Articles Next Articles

Cloning of garlic hexokinase gene AsHXK2 and expression analysis of its involvement in rhizosphere growth-promoting bacteria alleviating drought stress

GUO Chunqian¹(), TIAN Jie¹^,²^,^*()

1. Qinghai Key Laboratory of Vegetable Genetics and Physiology, Academy of Agriculture and Forestry Sciences, Qinghai University, Xining 810016, China
2. State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining 810016, China

Received:2021-08-24 Online:2022-09-25 Published:2022-09-30
Contact: TIAN Jie

Abstract

Abstract:

In order to study the alleviating role of AsHXK2 gene by plant growth promoting rhizobacteria under drought stress, TA cloning method was used to clone the sequence of AsHXK2 gene in Ledu purple garlic, and the basic characteristics were analyzed by bioinformatics. qRT-PCR was used to analyze the expression of AsHXK2 gene in different tissues of garlic, drought stress and rhizosphere growth promoting bacteria. The results showed that the open reading frame of AsHXK2 was 1 500 bp in length, encoding 499 amino acids. The molecular weight was about 57.77 ku, and the theoretical isoelectric point was 6.37. The subcellular localization prediction results showed that AsHXK2 protein was mainly located in chloroplast, containing a specific site, belonging to the hexose kinase 2 family. In terms of evolutionary relationship, garlic AsHXK2 was close to asparagus HXK2 in Asparagacea, qRT-PCR analysis showed that the expression of AsHXK2 gene had obvious tissue specificity, and the expression level in scale buds was the highest. In addition, AsHXK2 in different tissues showed different response patterns to drought stress induced by plant growth promoting rhizobacteria. Drought stress could significantly up-regulate AsHXK2 expression in roots, leaves and scale buds of garlic. However, in the treatment of rhizosphere growth-promoting bacteria and drought stress, the expression of AsHXK2 in leaves was significantly higher than that of drought stress, indicating that AsHXK2 in leaves was more obviously responsive to rhizosphere growth-promoting bacteria and drought stress. The study would provide the foundation for further exploring the mechanism of alleviating role by plant growth promoting rhizobacteria under drought stress.

Key words: garlic, hexokinase, gene cloning, expression analysis

CLC Number:

S633.4

GUO Chunqian, TIAN Jie. Cloning of garlic hexokinase gene AsHXK2 and expression analysis of its involvement in rhizosphere growth-promoting bacteria alleviating drought stress[J]. Acta Agriculturae Zhejiangensis, 2022, 34(9): 1925-1934.

Figures/Tables 9

Fig.1 AsHXK2 gene amplified by RT-PCR from garlic M, DNA marker (2 000 bp).

Fig.2 Nucleotide sequences and predicted amino acid sequences of AsHXK2 gene from garlic

Fig.3 Prediction of the transmembrane domain of AsHXK2 protein

Fig.4 Analysis of hydrophilicity and hydrophobicity of amino acid sequences of AsHXK2 from garlic

Fig.5 Secondary and tertiary structure prediction of AsHXK2 protein

Fig.6 Prediction of conserved domain of amino acid sequences of AsHXK2 from garlic

Fig.7 Phylogenetic tree of AsHXK2 amino acid sequence

Fig.8 AsHXK2 gene expression in different tissues of garlic The bars with different letters showed the significant difference (P<0.05). The same as below.

Fig.9 AsHXK2 gene expression in various tissues of garlic under different treatments A, Leaf; B, Root; C, Pseudostem; D, Scale bud.

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Cloning of garlic hexokinase gene AsHXK2 and expression analysis of its involvement in rhizosphere growth-promoting bacteria alleviating drought stress

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