橡胶树体细胞胚胎发生相关类受体蛋白激酶基因(HbSERK1)的克隆及表达分析

Cloning and Expression of HbSERK1 from Hevea brasiliensis

从橡胶树体细胞胚中克隆了一个与体细胞胚胎发生相关的类受体蛋白激酶基因,命名为HbSERK1。HbSERK1长2 159 bp,含有一个1 482 bp的阅读框,编码493个氨基酸。HbSERK1序列与毛果杨、葡萄、胡萝卜、马铃薯、拟南芥和燕麦的体细胞胚胎发生相关类受体蛋白激酶同源性分别为91%、90%、88%、88%、87%、86%。生物信息学分析表明,HbSERK1含有一个高度保守的蛋白激酶家族结构域,同时含有ATP结合区、蛋白激酶活化位点和Ser/Thr活性位点。RT-PCR半定量分析表明,HbSERK1在叶、胶乳、雄花、生长在愈伤诱导培养基上30 d和50 d的愈伤组织及生长在胚状体诱导培养基上12 d的胚性愈伤组织中均未表达,而生长在胚状体诱导培养基上21 d和40 d的胚性愈伤组织及成熟体细胞胚中表达。橡胶树HbSERK1基因主要在体细胞胚发生的早期表达,表明其可能参与橡胶树的体细胞胚早期发生过程。

The full-length cDNA encoding somatic embryogenesis receptor-like kinase, designated HbSERK1, was isolated from rubber tree. HbSERK1 was a 2 159 bp long containing a 1 482 bp open reading frame encoding a putative protein of 493 amino acids. The deduced amino acid sequences of HbSERK1 shared 91%, 90%, 88%, 88%, 87% and 86% identity with the somatic embryogenesis receptor-like kinases from Populus trichocarpa, Vitis vinifera, Daucus carota, Solarium tuberosum, Arabidopsis thaliana and Oryza sativa. HbSERK1 contained a conserved intracellular serine/threonine protein kinase domain. After about 20 days of culture on the embryo induction medium, HbSERK1 was expressed in calli, while HbSERK1 was little expressed in the calli growing onto calli induction medium, leaf and latex. It was suggested that HbSERK1 maybe take part in somatic embryogenesis during in vitro culture.