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| Biological Characteristics and Thermotolerance-regulating Roles of Wheat (Triticum aestivum) Heat Shock Transcription Factor Gene TaHsfA2f |
| ZHANG Yuan-Yuan1, 3, ZHAO Hui2, ZHANG Yu-Jie1, 4, DUAN Shuo-Nan1, LI Guo-Liang1, *, GUO Xiu-Lin1, * |
1 Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences / Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China;
2 College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang 050018, China;
3 College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China;
4 Hebei North University, Zhangjiakou 075000, China |
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Abstract Plant heat shock transcription factor (Hsf) is important regulating factor of signal transduction pathway activated transcriptively by genes under heat stress and other stresses.Hsf can specifically bind to heat shock elements (HSE) in the upstream promoter region of heat shock protein genes to realize the regulation to these genes expression.Plant Hsfs belong to multi-genes family, the members are different among varieties.Hsfs are divided into 3 classes of A, B and C, each class including several subclasses.Many previous works were mainly focused on class A, especially subclass A1 and A2.There are more than 56 members in wheat (Triticum aestivum) Hsf family reported in previous research, containing multiple subclass members, showing diverse characteristics and functions.Recently, the Hsf family members were found to reach to 82 based on wheat genome sequence.In this paper, The TaHsfA2f was isolated from wheat young leaves treated by heat shock at 37 ℃ for 1.5 h using homologous cloning methods.Sequence analysis showed that the coding sequence (CDS) of TaHsfA2f (GenBank No.MK045331) was 1 062 bp encoding a protein of 353 amino acids.The amino acid sequence analysis demonstrated that TaHsfA2f contained a DNA-binding domain (DBD), a nuclear localization signal (NLS) of MRKELEDAMSNKRRRR peptide, a nuclear export signal (NES) of LKRDKGLLM peptide and an aromatic, large hydrophobic and acidic amino residues (AHA) of DDFWEDLLHE peptide.Through transient reporter assays with tobacco (Nicotiana tabacum) epidermal cells, it was found that the TaHsfA2f protein was subcellular localized in the nuclei under normal growth conditions.Homologous analysis showed that TaHsfA2f protein shared higher identities with the HsfA2e proteins from some other crops such as barley (Hordeum vulgare), Sargassum mongolicus, Arabidopsis thaliana and so on.qRT-PCR analysis showed that TaHsfA2f was expressed in the majority of tissues and organs of wheat with higher expression level in mature roots and lower expression level in young shoots (P<0.05).TaHsfA2f expression in leaf were up-regulated by heat shock at 37 ℃, salicylic acid (SA) and H2O2 (P<0.05), respectively, and the peak values appeared at 60 min and 120 min after treatments.Through genetic transformation in wide type A.thaliana and assays of thermotolerances of transgenic line 10_19, line 13_9 and line 20_14 of overexpressing TaHsfA2f, the results revealed that overexpression of TaHsfA2f could improve both the basal and acquired thermotolerances of transgenic A.thaliana, and relative Hsp genes expression were upregulated to different degrees at 8 h after 2 different heat regimes treatments.Compared to WT, under heat stress, the seedlings growth potential of 3 transgenic lines were better and chlorophyll contents of the rosette leaves were higher and relative electric conductivity (REC) were lower, providing physiological evidences for phenotypes.These results could provide theoretical evidence for further understanding of biological characteristics and functions of subclass HsfA2 members of wheat Hsf family.
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Received: 12 October 2018
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Corresponding Authors:
myhf2002@163.com;guolianglili@163.com
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