华北农学报 ›› 2018, Vol. 33 ›› Issue (2): 14-19. doi: 10.7668/hbnxb.2018.02.003

所属专题: 小麦 生物技术

• 论文 • 上一篇    下一篇

小麦组氨酸磷酸转运蛋白TaHP4基因的克隆和表达分析

孙丽静1, 张哲2, 刘茜1, 胡梦芸1, 张颖君1, 吕亮杰1, 李辉1   

  1. 1. 河北省农林科学院 粮油作物研究所, 河北省作物遗传育种实验室, 河北 石家庄 050035;
    2. 西北农林科技大学, 陕西 杨凌 712100
  • 收稿日期:2017-11-20 出版日期:2018-04-28
  • 通讯作者: 李辉(1964-),男,河北石家庄人,研究员,博士,主要从事小麦遗传育种研究。
  • 作者简介:孙丽静(1982-),女,河北邯郸人,助理研究员,博士,主要从事小麦分子育种及分子生物学研究。
  • 基金资助:
    国家转基因生物新品种培育重大专项(2016ZX08002005-002);河北省现代农业产业技术体系小麦产业创新团队项目;河北省应用基础研究计划重点基础研究项目(13966305D);河北省财政专项(F14E055611)

Cloning and Expression Analysis of TaHP4 in Wheat

SUN Lijing1, ZHANG Zhe2, LIU Qian1, HU Mengyun1, ZHANG Yingjun1, LÜ Liangjie1, LI Hui1   

  1. 1. Institute of Cereal and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences, Hebei Provincial Laboratory of Crop Genetics and Breeding, Shijiazhuang 050035, China;
    2. Northwest A & F University, Yangling 712100, China
  • Received:2017-11-20 Published:2018-04-28

摘要: 细胞分裂素在植物生长发育和抵御盐、干旱等非生物胁迫过程中发挥着重要作用。为了研究小麦细胞分裂素信号转导途径中组氨酸磷酸转运蛋白基因的功能和在植物逆境响应中的分子机制,以电子克隆结合RT-PCR的方法,从小麦中克隆了组氨酸磷酸转运蛋白基因TaHP4。该基因的开放阅读框为456 bp,编码的蛋白长度为151个氨基酸,推测的蛋白分子量为17.71 ku,等电点为9.09,属于碱性蛋白,具有一个HPt结构域和一个负责磷酸基团传递的保守的组氨酸位点。多序列比对和系统进化树分析表明,TaHP4与水稻和玉米中不含保守组氨酸位点的OsPHP1-OsPHP3和ZmHP4亲缘关系较近,相似性为62.3%~88.7%;而与小麦中前期已经克隆的TaHP1-TaHP3亲缘关系较远,相似性仅为32.2%~34.2%。TaHP4基因表达模式具有较强的组织特异性,在叶、穗、茎等地上部表达量较高,而在根中则表达量很低。另外,TaHP4基因受盐、干旱和ABA抑制表达,初步推测该基因可能在小麦抵御逆境胁迫的生物学过程中具有重要作用。

关键词: 小麦, 细胞分裂素, 组氨酸磷酸转运蛋白, 基因克隆, 基因表达

Abstract: Cytokinin plays an important role in plant growth and development and resistance to salt and drought stresses. To study the function of histidine phosphotransfer proteins(HPs) in cytokinin signaling pathway and its molecular mechanism in plant stress response, TaHP4 was cloned in wheat based on electron cloning and RT-PCR. Sequence analysis showed that the open reading frame(ORF) of TaHP4 was 456 bp,encoding 151 amino acids of protein containing a HPt domain and a conserved histidine site responsible for the transmission of phosphate groups,with predicted molecular weight of 17.71 ku and isoelectric point of 9.09. Multiple sequence alignment and phylogenetic tree analysis showed that TaHP4 was closely related to OsPHP1-OsPHP3 and ZmHP4,with 62.3%-88.7% identity. However,TaHP4 was distantly related to the early cloned TaHP1-TaHP3,with only 32.2%-34.2% identity. The expression pattern of TaHP4 was highly tissue-specific,with higher expression in the leaf,panicle and stem,but less in root. In addition,the expression of TaHP4 was inhibited by salt,drought and ABA,suggesting that TaHP4 may play an important role in the biological process of wheat resistance to abiotic stress.

Key words: Wheat, Cytokinin, Histidine phosphotransfer proteins, Gene cloning, Gene expression

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引用本文

孙丽静, 张哲, 刘茜, 胡梦芸, 张颖君, 吕亮杰, 李辉. 小麦组氨酸磷酸转运蛋白TaHP4基因的克隆和表达分析[J]. 华北农学报, 2018, 33(2): 14-19. doi: 10.7668/hbnxb.2018.02.003.

SUN Lijing, ZHANG Zhe, LIU Qian, HU Mengyun, ZHANG Yingjun, LÜ Liangjie, LI Hui. Cloning and Expression Analysis of TaHP4 in Wheat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(2): 14-19. doi: 10.7668/hbnxb.2018.02.003.

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