小麦YSK<sub>2</sub>型脱水素基因<em>WDHN2</em>的克隆及表达分析

doi:10.3969/j.issn.1674-7968.2022.08.001

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Abstract Abiotic stress seriously affects the growth and yield of wheat (Triticum aestivum). Dehydrin (DHN) is a kind of protein induced by abiotic stress, and plays an important role in protecting cells from damage caused by water deficiency and temperature changes. In order to study the role of dehydrin in plants responding to abiotic stress, this study isolated a YSK₂ type dehydrin gene WDHN2 (GenBank No. OK094572) from wheat (Triticum aestivum) in homology-based cloning technology. WDHN2 gene had an open reading frame with the length of 501 bp, and encoded a protein containing 166 amino acids, which contained 1 segment of Y, 1 segment of S, and 2 segments of K. Phylogenetic analysis revealed that WDHN2 had the closest genetic relationship with Aegilops tauschii DHN (XP_020188302.1). The bioinformatics analysis showed that WDHN2 protein was a highly-hydrophilic protein, and its higher structure was mainly composed of random coil. Subcellular localization predicted that WDHN2 protein localized in the nucleus. In addition, S segment was the main phosphorylated site. The qPCR analysis indicated that the expression of WDHN2 gene was induced by drought, salinity, low temperature and abscisic acid (ABA). The prokaryotic fusion expression vector pET28a-WDHN2 was constructed and transformed into Escherichia coli BL21 (DE3). Then isopropyl β-D-1-thiogalactopyranoside (IPTG) was used for inducible expression, the 25 kD WDHN2 fusion protein and recombinant E. coli were obtained. Resistance analysis of recombinant bacteria showed that WDHN2 protein improved the tolerance to osmotic stress, high salinity, low temperature and high temperature stresses in Escherichia coli. The above results revealed that WDHN2 might be involved in the signal transduction pathway of wheat in response to abiotic stresses. The present study could enrich the research content of the molecular mechanism of abiotic stress responses in wheat, and provide basic material for further study on the function of WDHN2 gene.

Key words： Wheat Dehydrin (DHN) Bioinformatics analysis qPCR Procaryotic expression

Received: 28 October 2021

ZTFLH:

S512.1

Corresponding Authors: *zhanghongmei@haust.edu.cn

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	ZHANG Hong-Mei
	GONG Ming-Gui
	WU Jia-Fa
	ZHANG Min
	LIU Shuai
	XUE Hao-Bin
	CAO Zhi-Hao
	PAN Yong-Kang

Cite this article:

ZHANG Hong-Mei,GONG Ming-Gui,WU Jia-Fa, et al. Cloning and Expression Analysis of YSK₂ Type Dehydrin Gene WDHN2 in Wheat (Triticum aestivum)[J]. 农业生物技术学报, 2022, 30(8): 1443-1453.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2022.08.001 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2022/V30/I8/1443

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