玉米热激转录因子基因ZmHsf25的克隆、特性与耐热性功能分析

作物学报ACTA AGRONOMICA SINICA 2017, 43(7): 1021 1029https://www.360docs.net/doc/549610944.html,/

ISSN 0496-3490; CODEN TSHPA9E-mail: xbzw@https://www.360docs.net/doc/549610944.html, DOI: 10.3724/SP.J.1006.2017.01021

赵立娜1,2,**段硕楠1,**张华宁1郭秀林1,*李国良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 河北师范大学生命科学学院, 河北

石家庄 050024

摘要: 植物热激转录因子(heat shock transcription factor, Hsf)对下游热激蛋白基因及耐热性相关基因的表达起关键

调控作用。玉米中至少有30个Hsf家族成员, 其中B族有7个。在前期对玉米A1亚族热激转录因子基因ZmHsf06

克隆及特性、抗旱耐热性功能分析的基础上, 从玉米幼叶中克隆到B族热激转录因子基因ZmHsf25的完整编码序列,

序列全长957 bp, 编码318个氨基酸残基。蛋白质序列含有DNA结合结构域、核定位信号序列和核输出信号序列, 同

源分析结果显示, ZmHsf25与高粱Sb06g025710的相似性最高, 达92%。荧光定量分析表明, 正常生长条件下, ZmHsf25在玉米多个组织器官中表达, 幼嫩花粉中高表达, 幼嫩根系和雌穗中表达量较低。42℃热胁迫能显著上调根

系和叶片ZmHsf25的表达, 叶片中上升幅度高于根系, 处理50 min达到峰值; 正常条件下, 水杨酸和H2O2处理均使

根系和叶片ZmHsf25表达下调, 而热激条件下却表现为显著上调。通过在洋葱表皮细胞中瞬时表达并观察GFP荧光,

确定ZmHsf25定位在细胞核。通过转化酵母并进行热胁迫处理发现, 热胁迫同时降低正常和转ZmHsf25酵母的耐热

性, 但与转空载体对照相比, 转基因酵母的耐热性更强。推测ZmHsf25参与玉米热胁迫响应和花粉的发育过程, 可

能是水杨酸热激信号转导途径的下游组分。研究结果为进一步探明玉米热激转录因子家族成员的特性与功能提供了

依据。

关键词:玉米; 热激转录因子基因ZmHsf25; 表达; 亚细胞定位; 耐热性

Cloning, Characteristics and Regulating Role in Thermotolerance of Heat Shock Transcription Factor (ZmHsf25) in Zea mays L.

ZHAO Li-Na1,2,**, DUAN Shuo-Nan1,**, ZHANG Hua-Ning1, GUO Xiu-Lin1,*, and LI Guo-Liang1,*

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 Life Science, Hebei Normal University, Shijiazhuang 050024, China

Abstract: Heat shock transcription factors (Hsfs) are key components of signal transduction pathways involved in the activation

of genes in response to heat shock (HS) stress in plants. There are at least 30 Hsf members in maize and seven of which belong to class B. In our previous work, we obtained ZmHsf06, which belongs to subclass A1, and investigated the characteristics of expres-sion, subcellular localization, and regulating roles in thermotolerance and drought-stress tolerance of ZmHsf06. In the present study, ZmHsf25 was isolated from maize (Zea mays L.) young leaves treated with at 42°C for one hour using homologous cloning methods. The sequencing analysis showed that the coding sequence (CDS) of ZmHsf25 was 957 bp and encoded a protein of 318 amino acids. The amino acid sequence analysis demonstrated that ZmHsf25 contained a DNA-binding domain (DBD), a nuclear localization signal (NLS) of KRLR peptide and a nuclear export signal (NES) of VLTLSV peptide. The identity of amino acid between ZmHsf25 and Sb060g025710 of sorghum was the highest, which was 92%. ZmHsf25 was expressed in multiple tissues

本研究由河北省自然科学基金项目(C2017301065), 河北省博士基金项目(2017039349), 渤海粮仓科技工程项目(F16C14001)和河北省高层次人才项目(A201500130)资助。

This study was supported by the Natural Science Foundation of Hebei Province (C2017301065), the Doctoral Foundation of Hebei Province (2017039349), the Bohai Barn Science and Technology Project (F16C14001) and the High-level Talent Project of Hebei Province (A201500130).

*通讯作者(Corresponding authors): 李国良, E-mail: guolianglili@https://www.360docs.net/doc/549610944.html,, Tel: 0311-********; 郭秀林, E-mail: myhf2002@https://www.360docs.net/doc/549610944.html,, Tel: 0311-********

**同等贡献(Contributed equally to this work)

Received(收稿日期): 2016-11-08; Accepted(接受日期): 2017-03-02; Published online(网络出版日期): 2017-03-17.

URL: https://www.360docs.net/doc/549610944.html,/kcms/detail/11.1809.S.20170317.1938.004.html