橡胶树WRKY家族转录因子HbWRKY75基因的克隆及表达分析

陈晓丽1,2, 闫栋2,3, 孙丽娜2, 曾日中2,*, 杨礼富2,*
1山东大学生命科学学院, 济南250100; 2中国热带农业科学院橡胶研究所, 农业部橡胶树生物学与遗传资源利用重点开放实验室, 省部共建国家重点实验室培育基地, 海南儋州571737; 3海南大学农学院, 海口570228

通信作者:曾日中;E-mail: hnzrz@aliyun.com; ylfri@126.com

摘 要:

从巴西橡胶树无性系PR107的叶片组织中克隆了1个新的WRKY家族转录因子基因, 命名为HbWRKY75, 其ORF序列长为603 bp, 编码201个氨基酸。生物信息学分析表明, HbWRKY75在N端第110~160氨基酸之间含有1个典型的WRKY结构域, 属II类WRKY转录因子家族成员, 与葡萄、大豆、油棕、陆地棉和拟南芥等同类基因序列的同源性分别为92%、85%、71%、63%和57%。实时荧光定量RT-PCR分析表明, HbWRKY75基因在橡胶树叶片组织中的表达量最高, 而在叶柄组织中的表达量最低; HbWRKY75基因在橡胶树衰老晚期叶片中的表达量高于其他叶片发育时期; 乙烯利、茉莉酸甲酯(MeJA)、聚乙二醇 (PEG)、H2O2、NaCl、低温处理及机械损伤均可不同程度地诱导HbWRKY75基因的上调表达; 但ABA处理则诱导HbWRKY75基因的下调表达。HbWRKY75可能是与橡胶树叶片衰老和响应乙烯等激素刺激相关的转录因子, 并在橡胶树响应NaCl和低温等逆境胁迫中可能发挥调控作用。

关键词:巴西橡胶树; 转录因子HbWRKY75; 基因表达; 激素处理; 逆境胁迫

收稿:2015-10-28   修定:2016-02-02

资助:国家自然科学基金(31371556)。

Cloning and expression analysis of HbWRKY75 gene in leaf from Hevea brasiliensis

CHEN Xiao-Li1,2, YAN Dong2,3, SUN Li-Na2, ZENG Ri-Zhong2,*, YANG Li-Fu2,*
1College of Life Sciences, Shangdong University, Ji’nan 250100, China; 2Key Laboratory of Ministry of Agriculture for Rubber Tree Biology and Genetic Resource Utilization, State Key Laboratory Incubation Base, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences (CATAS), Danzhou, Hainan 571737, China; 3College of Agriculture, Hainan University, Haikou 570228, China

Corresponding author: ZENG Ri-Zhong; E-mail: hnzrz@aliyun.com; ylfri@126.com

Abstract:

A novel WRKY gene was cloned by RT-PCR from the leaf tissue of Hevea brasiliensis (para rubber tree) and designated as HbWRKY75. The open reading frame (ORF) of HbWRKY75 gene was 603 bp encoding a protein with 201 amino acid residues. The HbWRKY75 had a typical WRKY domain from amino acid 110 to 160 and belonged to the class II protein of the WRKY transcription factor family. The homology analysis revealed that HbWRKY75 shared amino acid sequence identities of 92%, 85%, 71%, 63% and 57% with the WRKY75 protein from Vitis vinifera, Zea mays, Elaeis guineensis, Nelumbo nucifera, and Gossypium hirsutum. The expression pattern of HbWRKY75 gene was investigated using real-time quantitative PCR, and the results demonstrated that the transcript level of HbWRKY75 gene in leaves was significantly higher than those in other parts of rubber trees. HbWRKY75 gene was differentially expressed in leaves with different developmental stages, and mostly expressed in the leaf at the late stage of senescence. The expression level of HbWRKY75 gene in the young leaves greatly increased compared with those of the control samples of rubber trees treated with Ethephon, MeJA (methyl jasmonic acid), PEG (polyethylene glycol), H2O2, NaCl, low temperature and bark tapping (mechanical damage); but ABA treatment down-regulated the expression of HbWRKY75. The presented data suggested that HbWRKY75 may be a transcription factor involved in the leaf senescence and the responses to phytohormones such as ethylene etc., and play a regulatory role in response to abiotic stresses such as NaCl and low temperature of rubber trees.

Key words: Hevea brasiliensis; HbWRKY75; gene expression; phytohormone; abiotic stress

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