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类钙调素蛋白(Calmodulin-like proteins,CMLs)在植物钙信号转导中发挥重要作用。本研究采用反转录PCR(reverse transcription PCR,RT-PCR)方法,从巴西橡胶树中克隆了CML基因Hb CML27。Hb CML27开放阅读框长度为450 bp,编码149个氨基酸。预测Hb CML27蛋白分子量为16.55 k D,等电点为4.10,是一个亲水蛋白。Hb CML27蛋白含有4个保守的EF-hand钙结合结构域,与其它植物CML27蛋白具有较高的同源性。实时荧光定量PCR分析表明,在巴西橡胶树叶片、花、根、树皮及胶乳等组织中均有Hb CML27基因表达,其中,表达量最低的组织是叶片,表达量最高的组织是胶乳。同健康橡胶树胶乳相比,Hb CML27在死皮橡胶树胶乳中的表达量降低。伤害、过氧化氢(H2O2)、乙烯利、茉莉酸甲酯(Me JA)、低温及干旱等处理均能上调Hb CML27的表达。这些结果表明Hb CML27可能参与了巴西橡胶树产排胶调控及响应非生物逆境胁迫。
Calmodulin-like proteins (CMLs) play an important role in plant calcium signaling. In this study, CML gene Hb CML27 was cloned from the rubber tree of Brazil by reverse transcription PCR (RT-PCR). Hb CML27 open reading frame length of 450 bp, encoding 149 amino acids. It is predicted that the molecular weight of Hb CML27 protein is 16.55 kD and its isoelectric point is 4.10, which is a hydrophilic protein. Hb CML27 protein contains four conserved EF-hand calcium binding domains, which have high homology with other plant CML27 proteins. Real-time quantitative PCR analysis showed that there was Hb CML27 gene expression in the leaves, flowers, roots, bark and latex of the rubber tree. Among them, the tissue with the lowest expression level was the leaf and the highest expression level was the latex. The expression level of Hb CML27 in dead-skin rubber latex was lower than that of healthy rubber latex. Injury, hydrogen peroxide (H2O2), ethephon, methyl jasmonate (Me JA), hypothermia and drought could up-regulate the expression of Hb CML27. These results indicate that Hb CML27 may be involved in the regulation of rubber gum production and response to abiotic stress.