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2025年4月6日 星期日
农业生物技术学报  2019, Vol. 27 Issue (5): 825-835    DOI: 10.3969/j.issn.1674-7968.2019.05.006
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
小麦热激转录因子基因TaHsfA2f生物学特性及耐热性调控作用
张园园1, 3, 赵慧2, 玉杰1, 4, 段硕楠1, 李国良1, *, 郭秀林1, *
1 河北省农林科学院 遗传生理研究所/河北省植物转基因中心,石家庄 050051;
2 河北科技大学 生物科学与工程学院,石家庄050018;
3 河北师范大学 生命科学学院,石家庄 050024;
4 河北北方学院,张家口 075000
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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摘要 植物热激转录因子(heat shock transcription factor, Hsf)在植物体响应热胁迫和其他逆境胁迫信号转导通路中发挥重要的调节作用,数量因作物不同差异较大。小麦(Triticum aestivum) Hsf家族成员数量多,包括多个亚家族,生物学特性和功能复杂多样。本研究从小麦中同源克隆了A2亚族成员TaHsfA2f(GenBank No.MK045331)的完整开放阅读框序列,序列长1 062 bp,编码353个氨基酸,蛋白质序列含DNA结合结构域(DNA-binding domain, DBD)、核定位信号序列(nuclear localization signal, NLS)(MRKELEDAMSNKRRRR)、核输出信号序列(nuclear export signal, NES)(LKRDKGLLM)和激活域(aromatic, large hydrophobic and acidic amino residues, AHA)(DDFWEDLLHE)。通过转化烟草(Nicotiana tabacum)表皮细胞发现,正常条件下TaHsfA2f蛋白质定位在细胞核。同源分析表明,TaHsfA2f与多种作物的HsfA2e蛋白相似性较高。组织特异性表达分析表明,TaHsfA2f在小麦中组成型表达。显著性分析表明,成熟根中高表达,幼茎和幼叶中较低表达(P<0.05)。叶片中TaHsfA2f的表达分别受热胁迫、水杨酸和过氧化氢处理上调(P<0.05),于处理60 min和120 min时达峰值。TaHsfA2f可不同程度提高转基因拟南芥(Arabidopsis thaliana)植株的基础耐热性和获得耐热性,在热胁迫条件下上调一系列热相关蛋白基因的表达。本研究为进一步解析小麦A2亚族基因特性与调控功能提供理论依据。
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张园园
赵慧
玉杰
段硕楠
李国良
郭秀林
关键词 小麦TaHsfA2f生物学特性亚细胞定位耐热性调控    
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.
Key wordsWheat TaHsfA2f    Biological characteristics    Subcellular-localization    Thermotolerance regulation
收稿日期: 2018-10-12     
ZTFLH:  S512.1  
基金资助:河北省自然科学基金重点项目(No.C2016301085)、河北省现代农业科技创新工程项目(494-0402-YBN-S2XB, 494-0402-YBN-C7GQ)和河北省高层次人才项目(No.A201500130)
通讯作者: myhf2002@163.com;guolianglili@163.com   
引用本文:   
张园园, 赵慧, 玉杰, 段硕楠, 李国良, 郭秀林. 小麦热激转录因子基因TaHsfA2f生物学特性及耐热性调控作用[J]. 农业生物技术学报, 2019, 27(5): 825-835.
ZHANG Yuan-Yuan, ZHAO Hui, ZHANG Yu-Jie, DUAN Shuo-Nan, LI Guo-Liang, GUO Xiu-Lin. Biological Characteristics and Thermotolerance-regulating Roles of Wheat (Triticum aestivum) Heat Shock Transcription Factor Gene TaHsfA2f. 农业生物技术学报, 2019, 27(5): 825-835.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.05.006     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I5/825
 
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