巴西橡胶树C型凝集素类受体激酶基因HbClecRLK1的克隆和表达分析  

Cloning and Expression Analysis of the C-type Lectin Receptor-like Kinase Gene HbClecRLK1 in Rubber Tree

作　　者：王梦希 田金华 罗红丽[1] Wang Mengxi;Tian Jinhua;Luo Hongli(College of Tropical Crops,Hainan University,Haikou,570228;Zhengzhou Urban River Management Office,Zhengzhou,450000)

机构地区：[1]海南大学热带作物学院,海口570228 [2]郑州市城区河道管理处,郑州450000

出　　处：《分子植物育种》2021年第20期6682-6689,共8页Molecular Plant Breeding

基　　金：海南省基础与应用基础研究计划(自然科学领域)高层次人才项目(2019RC071)资助。

摘　　要：类受体蛋白激酶(receptor-like protein kinases,RLKs)在植物的生长发育以及应答各种微生物侵染和不同环境胁迫的信号传递途径中发挥着重要作用。为初步探究C型凝集素类受体激酶基因家族在巴西橡胶树中的功能,本研究在橡胶树转录组测序基础上,通过RT-PCR方法扩增获得1个C型凝集素类受体激酶基因,命名为HbClecRLK1。该基因包含1个1626 bp的完整开放阅读框,编码一个541个氨基酸的多肽。生物信息学分析结果表明,HbClecRLK1蛋白包括3个保守结构域:凝集素类识别结构域、跨膜结构域和酪氨酸激酶结构域。氨基酸序列同源比对结果表明,橡胶树Hb ClecRLK1蛋白的氨基酸序列与木薯(Manihot esculenta Crantz)、蓖麻(Ricinus communis L.)、麻疯树(Jatropha curcas L.)的C型凝集素类受体激酶蛋白高度同源,同源性分别为91.34%、86.63%、86.97%。系统进化树分析结果显示,HbClecRLK1与其他物种的C型凝集素类受体激酶位于同一分支。亚细胞定位结果表明,Hb ClecRLK1蛋白位于细胞膜上。实时荧光定量PCR结果表明,当橡胶树叶片受到胶胞炭疽菌(Colletotrichum gloeosporiodes)和白粉病菌(Oidium heveae Steinm)侵染时,基因表达量明显升高。以上结果暗示Hb ClecRLK1基因可能参与橡胶树对白粉病和炭疽病的抗病反应,为进一步研究该基因的功能和寻找新的植物病害防控策略提供参考。Receptor-like protein kinases(RLKs) play important roles in plant growth and development as well as the signal transduction pathways in response to multiple biotic and abiotic stresses.In order to investigate the function of C-type lectin receptor kinases gene family in Hevea brasiliensis,a C-type lectin receptor-like kinase gene named HbClecRLK1 was amplified by RT-PCR based on the RNA-Seq database of rubber tree.This gene contains a complete open reading frame of 1 626 bp,which encodes a polypeptide of 541 amino acids.The results of bioinformatics showed that the Hb ClecRLK1 protein had a C-type lectin domain,a transmembrane domain and a tyrosine kinase domain,and the amino acid sequence of Hb ClecRLK1 was 91.34%,86.63% and 86.97%,which is similar to that of ClecRLK protein from Manihot esculenta Crantz,Ricinus communis L.and Jatropha curcas L.,respectively.Phylogenetic tree results showed that Hb ClecRLK1 was located in the same branch with the known ClecRLKs from other species.The subcellular localization results showed that the HbClecRLK1 protein was located on the cell membrane.Real-time quantitative PCR results showed that the expression of HbClecRLK1 was induced significantly in rubber trees leaves inoculated with Colletotrichum gloeosporioides and Odium heveae Steinm.The above results suggested that the HbClecRLK1 may be involved in the resistance of rubber tree to C.gloeosporiodes and O.heveae Steinm.The results will provide reference for further research on the function of HbClecRLK1 gene and search for new plant disease prevention and control tactics.

关 键 词：巴西橡胶树 C型凝集素类受体激酶 HbClecRLK1 表达分析 抗病反应 

分 类 号：S794.1[农业科学—林木遗传育种]