小麦热激转录因子基因TaHsfA2e特性及耐热性功能初探

doi:10.3724/SP.J.1006.2018.01818

作物学报 ›› 2018, Vol. 44 ›› Issue (12): 1818-1828.doi: 10.3724/SP.J.1006.2018.01818

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

小麦热激转录因子基因TaHsfA2e特性及耐热性功能初探

张玉杰^1,²,张园园^1,³,张华宁¹,秦宁^1,²,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重点实验室, 河北石家庄 050051
2河北北方学院, 河北张家口 075000
3河北师范大学生命科学学院, 河北石家庄 050024

收稿日期:2018-04-19 接受日期:2018-07-20 出版日期:2018-12-12 网络出版日期:2018-08-02
通讯作者: 李国良,郭秀林
基金资助:
本研究由国家重点研发计划项目(2017YFD0300408);河北省自然科学基金重点项目(C2016301085);河北省现代农业科技创新工程项目(494-0402-YBN-RDC4);河北省现代农业科技创新工程项目(494-0402-YBN-SVE2);河北省高层次人才项目资助(A201500130)

Characterization and Regulatory Roles in Thermotolerance of Wheat Heat Shock Transcription Factor Gene TaHsfA2e

Yu-Jie ZHANG^1,²,Yuan-Yuan ZHANG^1,³,Hua-Ning ZHANG¹,Ning QIN^1,²,Guo-Liang LI^1,^*(),Xiu-Lin GUO^1,^*()

1 Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences / Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, Hebei, China
2 Hebei North University, Zhangjiakou 075000, Hebei, China
3 College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, Hebei, China

Received:2018-04-19 Accepted:2018-07-20 Published:2018-12-12 Published online:2018-08-02
Contact: Guo-Liang LI,Xiu-Lin GUO
Supported by:
This study was supported by the National Key Research and Development Program of China(2017YFD0300408);the Key Project of Natural Science Foundation of Hebei Province(C2016301085);the Science and Technology Innovation Program for Modern Agriculture in Hebei Province(494-0402-YBN-RDC4);the Science and Technology Innovation Program for Modern Agriculture in Hebei Province(494-0402-YBN-SVE2);the High-level Talent Project of Hebei Province(A201500130)

摘要/Abstract

摘要：

植物热激转录因子(heat shock transcription factor, Hsf)能够通过激活热激蛋白基因的表达而启动热激反应, 在传递热信号以及提高植物耐热性方面发挥重要的调控作用。植物Hsf属多基因家族, 数量因作物不同差异较大。小麦Hsf家族成员多, 特性和功能复杂多样。本文在通过生物信息学推测分析小麦Hsf家族基因数目及其分类的基础上, 从小麦幼叶中同源克隆获得A2亚族成员TaHsfA2e (GenBank登录号为MG700614)的完整编码序列, 序列长1026 bp, 编码341个氨基酸残基, 含完整的DNA结合结构域DBD、核定位信号序列NLS、核输出信号序列NES和激活域AHA。正常条件下TaHsfA2e蛋白质被定位在细胞核。同源分析表明, TaHsfA2e与小麦HsfA6f蛋白相似性最高, 达96%。定量分析表明, TaHsfA2e在小麦多个组织器官中表达量均低, 但在成熟种子中高表达。叶片中TaHsfA2e的表达受37℃热胁迫显著上调, 处理60 min时达峰值, 被SA和H₂O₂下调。50℃热胁迫下转TaHsfA2e酵母细胞的耐热性显著强于转空载体对照, TaHsfA2e可不同程度提高转基因拟南芥植株的基础耐热性和获得耐热性, 上调热胁迫条件下热相关蛋白基因的表达。

关键词: TaHsfA2e, 表达特性, 亚细胞定位, 遗传转化, 耐热性

Abstract:

As key regulatory genes in the signal pathway responsive to heat stress, plant heat shock transcription factors (Hsfs) can enhance plant thermotolerances by triggering hsp or other relative genes to express. Plant Hsfs belong to multi-genes family, the members are different among varieties. Based on the phylogenetic tree of Hsf proteins from wheat (Triticum aestivum), rice (Oryza sativa) and Arabidopsis, we isolated the TaHsfA2e (GenBank accession number MG700614) from wheat young leaves treated with 37°C for 1.5 h using homologous cloning methods. Sequence analysis showed that the coding sequence (CDS) of TaHsfA2e is 1026 bp in length and encodes 341 amino acid residues. The TaHsfA2e protein was predicted to contain a DNA-binding domain (DBD), a nuclear localization signal (NLS) of KRRRP peptide, a nuclear export signal (NES) of LENLAMNI peptide and an aromatic, large hydrophobic and acidic amino residues (AHA) of CCFWEELLSE peptide, and localized in the nuclei under normal growth conditions. TaHsfA2e shared 96%, 94%, and 94% identities with HsfA6f and HsfA2d from wheat and HsfA2d from Aegilops tauschii, respectively. TaHsfA2e was lowly expressed in majority of tissues and organs but highly expressed in mature seeds of wheat, and the gene expression in leaf was up-regulated by heat shock at 37°C, with the peak value at 60 min after treatment, but down-regulated by salicylic acid or H₂O₂. TaHsfA2e could be induced by Gal in yeast (Saccharomyces cerevisiae), and yeast overexpressing pYES2-TaHsfA2e showed stronger growth potential than the controls expressing pYES2 after heat shock at 50 °C for 45 min, though all of the yeast growth potential were also decreased after treatment. Both of basal and acquired thermotolerances of transgenic Arabidopsis plants that overexpressed TaHsfA2e were improved, and the expressions of Hsp genes were up-regulated to different degrees. These results are essential for deep understanding biological functions and regulatory mechanism of subclass A2 Hsf members in plants.

Key words: TaHsfA2e, expression characterization, subcellular-localization, genetic transformation, thermotolerance

张玉杰,张园园,张华宁,秦宁,李国良,郭秀林. 小麦热激转录因子基因TaHsfA2e特性及耐热性功能初探[J]. 作物学报, 2018, 44(12): 1818-1828.

Yu-Jie ZHANG,Yuan-Yuan ZHANG,Hua-Ning ZHANG,Ning QIN,Guo-Liang LI,Xiu-Lin GUO. Characterization and Regulatory Roles in Thermotolerance of Wheat Heat Shock Transcription Factor Gene TaHsfA2e[J]. Acta Agronomica Sinica, 2018, 44(12): 1818-1828.

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链接本文: https://zwxb.chinacrops.org/CN/10.3724/SP.J.1006.2018.01818

https://zwxb.chinacrops.org/CN/Y2018/V44/I12/1818

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基因 Gene	正向序列 Forward sequence (5°-3°)	反向序列 Reverse sequence (5°-3°)
AtHsp18.2	GCAGATTAGCGGAGAGAGGA	CCTTCACTTCTTCCATCTTTGC
AtHsp21	AAGTCCGCTACACCGTTCTC	CCAACAATCCGAAAGGAGAG
AtHsfa32	GCGAAGTTGGTTGAGTGGTT	GGAGGAACTGAGAACAGATTGG
AtERDJ3A	CTCCTGTTTGTATCATTGGTGC	TGTGTCCTGAGAACCTGTGG
AtHsp25.3	GACGTCTCTCCTTTCGGATTGT	CTCCACTTCCTCCTCTGTTTCTTC
AtHsp70T	TGATTGAGGTGAGGATGCC	CCACTTCAACGACAAACCC
AtHsp90	CCCTCTCTTCTTCATAAATCAACA	CCATCGCAACGAACTTTG
AtHsp101	TGTCTTCAACACTCTGCTCCA	CACTTCCATTGTTACTTTCCCAG

小麦热激转录因子基因TaHsfA2e特性及耐热性功能初探

Characterization and Regulatory Roles in Thermotolerance of Wheat Heat Shock Transcription Factor Gene TaHsfA2e

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