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首页> 中文期刊> 《作物学报》 >玉米热激转录因子基因ZmHsf25的克隆、特性与耐热性功能分析

玉米热激转录因子基因ZmHsf25的克隆、特性与耐热性功能分析

         
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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.%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℃ 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 and organs of maize, and transcription expression level of ZmHsf25 was the highest in pollens compared with root, stem, func-tional leaf, immature embryo and ear. qRT-PCR results showed that ZmHsf25 was up-regulated by 42℃ heat shock in both leaves and roots. Under normal conditions, ZmHsf25 was down-regulated by both salicylic acid CSA and H2O2, but significantly up-regulated by heat stress at 42℃. Through transient reporter assay with onion (Allium cepa L.) epidermal cells, we found that ZmHsf25 was localized in nuclei. ZmHsf25 overexpressed yeast showed stronger thermotolerance than the controls after heat shock (HS), though yeast thermotolerances were also decreased by HS. The results revealed that ZmHsf25 perhaps is one of downstream elements of SA signal pathway to play a key role in regulating the response to heat stress and pollen development. These results provide a theoretical basis for analyzing biological characteristics and functions of maize Hsf members further.

著录项

  • 来源
    《作物学报》 |2017年第7期|1021-1029|共9页
  • 作者

    赵立娜; 段硕楠; 张华宁; 郭秀林; 李国良;

  • 作者单位

    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;

    河北师范大学生命科学学院, 河北石家庄 050024;

    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;

    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;

    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;

    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;

  • 原文格式 PDF
  • 正文语种 chi
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  • 关键词

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

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