《植物生理学报》 2016, 52(3): 250-258
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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
Corresponding author: ZENG Ri-Zhong; E-mail: email@example.com; firstname.lastname@example.org
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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