橡胶树HbRPM1-3基因克隆及其应答白粉菌侵染的功能分析  

Cloning of HbRPM1-3 gene from Hevea brasiliensis and functional analysis of its response to powdery mildew infection

作　　者：柯宇航 刘明洋 王燕 梁晓宇[1,2,3] 王萌 张宇[1,2,3] KE Yuhang;LIU Mingyang;WANG Yan;LIANG Xiaoyu;WANG Meng;ZHANG Yu(School of Plant Protection,Hainan University,Haikou 570228,China;Natural Rubber Cooperative Innovation Center of Hainan Province&Ministry of Education of PRC,Haikou 570228,China;Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests(Hainan University),Ministry of Education,Haikou 570228,China)

机构地区：[1]海南大学植物保护学院,海口570228 [2]天然橡胶省部共建协同创新中心,海口570228 [3]热带农林生物灾害绿色防控教育部重点实验室,海口570228

出　　处：《植物生理学报》2021年第11期2167-2178,共12页Plant Physiology Journal

基　　金：国家自然科学基金(32160370和31860183);现代农业产业技术体系建设专项资金(CARS-33-BC1);海南省研究生创新科研课题(Hys2020-258)。

摘　　要：CNL(CC-NBS-LRR)蛋白是一种免疫受体蛋白,在植物抗病信号网络中起着重要的作用。利用PCR技术从橡胶树(Hevea brasiliensis)无性系‘热研73397’叶片中克隆得到HbRPM1-3基因全长cDNA序列,其开放阅读框长2823 bp,编码940个氨基酸。蛋白结构分析发现HbRPM1-3蛋白含有RX-CC-Like、NBARC和LRR保守结构域,属于CNL蛋白。系统进化分析发现该蛋白与木薯(Manihotesculenta)的CNL蛋白具有较近的亲缘关系。亚细胞定位显示HbRPM1-3基因定位在细胞膜和细胞核中。qRT-PCR结果表明,HbRPM1-3基因主要在橡胶树叶片中表达,其表达量在白粉菌侵染下显著上调。水杨酸(salicylic acid,SA)、茉莉酸甲酯(methyl jasmonate,MeJA)、乙烯利(ethrel,ETH)和过氧化氢(H2O2)均能诱导橡胶树叶片中的HbRPM1-3基因显著上调表达。本研究认为HbRPM1-3基因在白粉病病原体感染的先天免疫机制中发挥重要作用。CNL(CC-NBS-LRR) protein is an immune receptor protein,which plays an important role in plant disease resistance signaling network.The full-length cDNA sequence of Hb RPM1-3 gene was cloned from the leaves of rubber tree(Hevea brasiliensis) clone ’Reyan 73397’ by PCR technique,and the open reading frame(ORF) of Hb RPM1-3 was 2 823 bp long and encoded 940 amino acids.The protein structure analysis of HbRPM1-3 showed that the protein contained RX-CC-like,NB-ARC and LRR conserved domain,belonged to the CNL protein.Phylogenetic analysis showed that the protein was closely related to the CNL protein of Manihot esculenta.Subcellular localization showed that Hb RPM1-3 gene was localized in cell membrane and nucleus.Quantitative real-time PCR results showed that Hb RPM1-3 genes expressed mainly in the leaves of the rubber tree.The expression level of Hb RPM1-3 was significantly upregulated under powdery mildew infection.Salicylic acid(SA),methyl jasmonate(JA),ethylene(ETH) and hydrogen peroxide(H_(2)O_(2)) could induce significantly upregulated expression of Hb RPM1-3 gene in rubber tree leaves.The results suggested that Hb RPM1-3 gene plays an important role in the innate immune mechanism of powdery mildew pathogen infection.

关 键 词：CNL蛋白 HbRPM1-3基因 橡胶树白粉菌 表达分析 亚细胞定位 

分 类 号：S763.7[农业科学—森林保护学]