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| Cloning, Expression and Functional Analysis of WDHN1 Gene from Wheat (Triticum aestivum) |
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Abstract YSK2 type dehydrins (DHNs), which are the most types of DHNs, have been shown to be involved in plant response and adaptation to various abiotic stresses. In order to determine the function of YSK2 DHNs, WDHN1 from wheat (Triticum aestivum) was cloned. The CDS of WDHN1 had a length of 491 bp, contained 2 exons and 1 intron, encoding a protein of 133 amino acids, containing one Y segment, one S segment and two K segments. A phylogenetic tree analysis with the related DHNs from different plant species indicated that WDHN1 shared homology with DNA from Aegilops tauschii(EMT30992). Based on PLACE and PlantCARE database analysis, the abiotic stress-related elements of WDHN1 promoter were determined, which contained 2 ABRE (abscisic acid (ABA) response element), and 3 MBS (MYB binding site). qRT-PCR analysis indicated that transcript accumulation occurred in response to low temperature, NaCl, ABA and PEG 6000 treatments. Tissue specific expression analysis showed that the transcript levels of WDHN1 reached the highest level at 22 d in embryo. The prokaryotic expression vector of WDHN1 was successfully constructed. The expression of fusion protein was obtained with isopropyl β-D-1-thiogalactopyranoside (IPTG) and its relative molecular weight was 20 kD. We investigated the anti-aggregation effects of protein WDHN1 on Escherichia coli viability and lactic dehydrogenase (LDH) activity during multiple abiotic stress treatments. WDHN1 protein enhanced tolerance of E. coli and LDH stabilization against diverse stresses via anti-aggregation effects. The results revealed that WDHN1 might serve as a potential stress response gene for the improvement of wheat inbred lines and cultivars under stress conditions in breeding activity.
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Received: 11 May 2016
Published: 01 October 2016
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| Fund:Specialized Research Fund for the Doctoral Program of Higher Education |
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