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.
[1] 沈迎芳,马超,吴小培,等. 2016. 扁蓿豆SK2型脱水素基因MrDHN3的异源表达提高大肠杆菌对盐和高温胁迫的抗性[J]. 草业学报, 25(8): 118-127. (Shen Y F, Ma C, Wu X P, et al.2016. Heterologous expression of an SK2-type dehydrin gene (MrDHN3) from Medicago ruthenica enhances Escherichia coli tolerance under salt and high temperature stress[J]. Acta Prataculturae Sinica, 25(8): 118-127.) [2] 谢建平, 袁世力, 刘星辰, 等. 2018. 狗牙根品种C299脱水素基因抗逆功能分析[J].中国草地学报, 40(4): 16-22. (Xie J P, Yuan S L, Liu X C, et al.2018. Analysis of stress-resistant function of dehydrin gene from Bermudagrass cv. 299[J]. Chinese Journal of Grassland, 40(4): 16-22.) [3] Abedini R, GhaneGolmohammadi F, PishkamRad R, et al.2017. Plant dehydrins: Shedding light on structure and expression patterns of dehydrin gene family in barley[J]. Journal of Plant Research, 130: 747-763. [4] Agarwal T, Upadhyaya G, Halder T, et al.2017. Difffferent dehydrins perform separate functions in Physcomitrella patens[J]. Planta, 245(1): 101-118. [5] Alsheikh M K, Svensson J T, Randall S K.2005. Phosphorylation regulated ion-binding is a property shared by the acidic subclass dehydrins[J].Plant Cell and Environment, 28(9): 1114-1122. [6] Egerton-Warburton L M, Balsamo R A, Close T J.1997. Temporal accumulation and ultrastructural localization of dehydrins in Zea mays[J]. Physiologia Plantarum, 101(1): 61-74. [7] Fujita M,Fujita Y, Noutoshi Y, et al.2006. Crosstalk between abiotic and biotic stress responses: A current view from the points of convergence in the stress signaling networks[J]. Current Opinion in Plant Biology, 9(4): 436-442. [8] Graether S P, Boddington K F.2014. Disorder and function: A review of the dehydrin protein family[J]. Frontiers of Plant Science, 5: 576. [9] Halder T, Upadhyaya G, Ray S.2017. YSK2 type dehydrin (SbDhn1) from Sorghum bicolor showed improved protection under high temperature and osmotic stress condition[J]. Frontiers in Plant Science, 8(918): 1-17. [10] Hara M.2010.The multifunctionality of dehydrins: An overview[J]. Plant Signalling & Behavior, 5(5): 503-508. [11] Hernández-Sánchez I E, Maruri-López I, Ferrando A, et al.2015. Nuclear localization of the dehydrin OpsDHN1 is determined by histidine-rich motif[J]. Frontiers in Plant Science, 6(702): 1-8. [12] Jardak-Jamoussia R, Zarroukb O, Salema A B, et al.2016. Overexpressing Vitis vinifera YSK2 dehydrin in tobacco improves plant performance[J]. Agricultural Water Management, 164: 176-189. [13] Jing H,Li C, Ma F, et al.2016. Genome-wide identification, expression diversication of dehydrin gene family and characterization of CaDHN3 in pepper (Capsicum annuum L.)[J]. PLOS ONE, 11(8): 1-19. [14] Ju H, Li D, Li D, et al.2021. Overexpression of ZmDHN11 could enhance transgenic yeast and tobacco tolerance to osmotic stress[J]. Plant Cell Reports, 40(9): 1723-1733. [15] Kalemba E M, Zadworna A B, Ratajczak E.2014. Multiple subcellular localizations of dehydrin-like proteins in the embryonic axes of common beech (Fagus sylvatica L.) seeds during maturation and dry storage[J]. Journal of Plant Growth Regulation, 34: 137-149. [16] Kim E C, Lee H S, Choi D W.2012. Sequence variability and expression pattern of the dehydrin gene family in Populus alba×P. tremula var. glandulosa[J]. Plant Omics Journal, 5(2): 122-127. [17] Koag M C, Fenton R D, Wilkens S, et al.2003. The binding of maize DHN1 to lipid vesicles. Gain of structure and lipid specificity[J]. Plant Physiology, 131(1): 309-316. [18] Kosova K, Vitamvas P, Prasil I T.2014.Wheat and barley dehydrins under cold, drought, and salinity: What can LEA-Ⅱ proteins tell us about plant stress response[J]. Frontiers in Plant Science, 5: 343. [19] Li Q, Zhang X, Lv Q, et al.2017. Physcomitrella patens dehydrins (PpDHNA and PpDHNC) confer salinity and drought tolerance to transgenic Arabidopsis plants[J]. Frontiers in Plants Science, 8(1316): 1-15. [20] Liu H, Yang Y, Zhang L S.2019. Identification of upstream transcription factors and an interacting PP2C protein of dehydrin WZY2 gene in wheat[J]. Plant Signaling & Behavior, 14(12): 1678370. [21] Liu Y, Li D X, Song Q P, et al.2019. The maize late embryogenesis abundant protein ZmDHN13 positively regulates copper tolerance in transgenic yeast and tobacco[J]. Crop Journal, 7(3): 403-410. [22] Lv A, Fan N, Xie J, et al.2017. Expression of CdDHN4, a novel YSK2-type dehydrin gene from Bermudagrass, responses to drought stress through the ABA-dependent signal pathway[J]. Frontiers in Plant Science, 8(748): 1-13. [23] Maszkowska J, Janusz D, Kulik A, et al.2019. Phosphoproteomic analysis reveals that dehydrins ERD10 and ERD14 are phosphorylated by SNF1-related protein kinase 2.10 in response to osmotic stress[J]. Plant Cell and Environment, 42(3): 931-946. [24] Meng Y C, Zhang H F, Pan X X, et al.2021. CaDHN3, a pepper (Capsicum annuum L.) dehydrin gene enhances the tolerance against salt and drought stresses by reducing ROS accumulation[J]. International Journal Molecular Science, 22(3205): 1-18. [25] Nguyen P N, Tossounian M A, Kovacs D S, et al.2020. Dehydrin ERD14 activates glutathione transferase Phi9 in Arabidopsis thaliana under osmotic stress[J]. Biochimica et Biophysica Acta-General Subjects, 1864(3): 129506. [26] Nylande M, Svensson J, Palva E T, et al.2001. Stress-induced accumulation and tissue-specifific localization of dehydrins in Arabidopsis thaliana[J]. Plant Molecular Biology, 45: 263-279. [27] Palmer S R, De V R, Graether S P.2019. Sequence composition versus sequence order in the cryoprotective function of an intrinsically disordered stress-response protein[J]. Protein Science, 28(8): 1448-1459. [28] Perdiguero P, Collada C, Soto A.2014. Novel dehydrins lacking complete K-segments in Pinaceae. The exception rather than the rule[J]. Frontiers in Plant Science, 5(682): 1-9. [29] Romanenko A,Borovskii G,Ukolova I,et al.2010. Subcellular localization of dehydrin in wheat plants subjected to low-temperature adaptation[J]. Biologicheskie Membrany, 27(2): 156-165. [30] Saibi W, Zouari N, Masmoudi K, et al.2016. Role of the durum wheat dehydrin in the function of proteases conferring salinity tolerance in Arabidopsis thaliana transgenic lines[J]. International Journal of Biological Macromolecules, 85: 311-316. [31] Ucarli C, McGuffin L J, Caputlu S, et al.2016. Genetic diversity at the Dhn3 locus in Turkish Hordeum spontaneum populations with comparative structural analyses[J]. Scientific Reports, 6(20966): 1-10. [32] Yang Y, He M, Zhu Z, et al.2012. Identification of dehydrin gene family from grapevine species and analysis of their responsiveness to various forms of abiotic and biotic stress[J]. BMC Plant Biology, 12: 140. [33] Zhang H, Zheng J, Su H, et al.2018. Molecular cloning and functional characterization of the dehydrin (IpDHN) gene from Ipomoea pes-caprae[J]. Frontiers in Plant Science, 9: 1454.