漾濞大泡核桃病程相关蛋白10基因JsPR10-1的克隆与表达分析  

Cloning and Expression Analysis of a Pathogenesis-related Protein 10 Gene from Juglans sigillata

作　　者：何华[1] 陈朝银[1] 韩青[1] 张南南[1] 葛锋[1] 刘迪秋[1] 

机构地区：[1]昆明理工大学生命科学与技术学院,昆明650500

出　　处：《植物科学学报》2014年第6期612-619,共8页Plant Science Journal

基　　金：国家科技部科技支撑计划项目(2011BAD46B00)

摘　　要：病程相关蛋白(pathogenesis-related protein,PR)10的激活与积累在植物抗逆境胁迫中有非常重要的作用。根据漾濞大泡核桃(Juglans sigillata)编码PR10的EST(expressed sequence tag)序列设计引物,利用快速扩增c DNA末端技术,克隆得到PR10基因的全长c DNA序列,并命名为Js PR10-1。Js PR10-1全长c DNA为776 bp,含有483 bp的开放阅读框、74 bp 5'-非编码区以及219 bp 3'-非编码区,编码含有160个氨基酸的蛋白质。全长基因序列中含有1个124 bp的内含子。Js PR10-1编码的蛋白质与栎树(Quercus suber)、欧洲山毛榉(Fagus sylvatica)以及欧洲板栗(Castanea sativa)的PR10相似性较高,并且在PR10的系统进化树中与双子叶植物聚为一支。qRT-PCR分析结果表明,植物信号分子水杨酸、茉莉酸、乙烯以及过氧化氢处理均可诱导Js PR10-1表达。在接种胶孢炭疽菌(Colletotrichum gloeosporioides)后,Js PR10-1的表达量迅速上升并在接种8 h时达到最大值,表明Js PR10-1参与漾濞大泡核桃对胶孢炭疽菌的防卫反应。本研究为揭示漾濞大泡核桃抗性机制奠定了理论依据。The activation and accumulation of pathogenesis-related protein （PR）10 play predominant roles in the defense response against pathogens. Based on a Juglans sigillata expressed sequence tag lEST） encoding PR10, gene-specific primers were designed and used to amplify the full-length cDNA of a novel PR10 gene by rapid amplification of cDNA ends （RACE）. This gene was named as JsPR10-1. JsPR10-1 was 776 bp in length with an open reading frame （ORF） of 483 bp, a 5＇-untranslated region （UTR） of 74 bp, and a 3＇-UTR of 219 bp, and the ORF encoded a protein with 160 amino acid residues. There was an intron of 124 bp in the genomic sequence of JsPRIO-1. Homology analysis indicated that JsPR10-1 was homologous with PR10s from Quercus suber, Fagus sylvatica and Castanea sativa; moreover, JsPR10-1 clustered together with the PR10s from dicots in the phylogenetic tree. qRT-PCR analysis showed that the expression levels of JsPR10-1 were induced by four different plant signaling molecules, including salicylic acid, jasmonic acid, ethylene, and H202. In addition, after inoculation with Colletotrichum gloeosporioides, JsPR10-1 was sharply up-regulated with the highest expression level at 8 h. The JsPRIO-1 gene was involved in the defense response against C. gloeosporioides. This experiment lays a theoretical foundation for revealing the mechanism of resistance in J. sigillata.

关 键 词：漾濞大泡核桃 病程相关蛋白10 基因克隆 胶孢炭疽菌 防卫反应 

分 类 号：Q78[生物学—分子生物学]