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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)、H_2O_2、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。
A new WRKY family transcription factor gene was cloned from leaves of clonal PR107, named HbWRKY75. Its ORF sequence was 603 bp in length and encoded 201 amino acids. Bioinformatics analysis showed that HbWRKY75 contains a typical WRKY domain between the 110th to the 160th amino acids of N-terminus, which is a member of class II WRKY transcription factor family, which is related to grapes, soybean, oil palm, upland cotton and Arabidopsis thaliana Homology of similar gene sequences were 92%, 85%, 71%, 63% and 57%, respectively. Real-time quantitative RT-PCR analysis showed that the expression level of HbWRKY75 gene was the highest in rubbery leaves and lowest in petiole tissues. The expression level of HbWRKY75 in late leaves of rubbery trees was higher than that in other leaf development stages. MeJA, PEG, H 2 O 2, NaCl, low temperature treatment and mechanical injury all induced HbWRKY75 up-regulated expression to some extent. However, down-regulated expression of HbWRKY75 gene was induced by ABA treatment. HbWRKY75 may be a transcription factor related to senescence and response to ethylene and other hormone stimuli in rubber leaves and may play a regulatory role in rubber tree response to stress such as NaCl and low temperature.