三七病程相关蛋白PR10-1基因克隆及功能初步分析  

Cloning and functional characterization of pathogenesis-related PR10-1 gene in Panax notoginseng

作　　者：唐美琼[1] 闵丹丹[2] 李刚[1] 蒋妮[1] 叶云峰[1] 

机构地区：[1]广西药用植物园,广西药用资源保护与遗传改良重点实验室,广西南宁530023 [2]广西中医药大学,广西南宁530000

出　　处：《药学学报》2015年第2期227-232,共6页Acta Pharmaceutica Sinica

基　　金：广西科学研究与技术开发项目(桂科重1298001-1-4);广西药用植物园青年基金项目(桂药基20);广西自然科学基金项目(2013GXNSFBA019087)

摘　　要：采用同源克隆法和RACE技术相结合,对三七根部蛋白质组差异进行研究,鉴定PR10-1蛋白相应的基因全长c DNA,初步分析该基因在三七中的功能。测序结果显示该基因序列全长863 bp,包含1个序列长465 bp编码155个氨基酸的开放阅读框,命名为Pn PR10-1,Gen Bank登录号KJ741402。氨基酸序列同源性及系统发育树分析发现,Pn PR10-1与人参的PR10-1蛋白同源性最高,含有Bet-v-I家族等多个保守结构域。实时荧光定量PCR分析显示,Pn PR10-1在1~3年生三七各组织中均有本底表达,暗示其参与生长发育、代谢调控等生物学过程;Pn PR10-1在根部受根腐病原菌（Fusarium oxysporum）胁迫上调表达,推测Pn PR10-1基因可能参与抗三七根腐病等广谱抗病防御反应。With homology cloning approaches coupling with RACE（rapid-amplification of c DNA ends） techniques, the full-length coding sequence of pathogenesis-related protein PR10-1 with differential expression was cloned from the total RNA of the root of Panax notoginseng, and its function was explored furtherly. As a result, the longest 465 bp ORF（named as Pn PR10-1 with the Accession No. KJ741402 in Gen Bank） was detected from the cloned sequence with full-length of c DNA of 863 bp. The corresponding peptide encoded consisted of 155 amino acids, contained some domains such as Bet-v-I, and showed high similarity with that from Panax ginseng by analysis of phylogenetic trees created from the alignments. Real-time quantitative PCR showed that the expression of Pn PR10-1 gene was constitutive in different tissues of 1-3 year old plant, suggesting that it might be involved in growth, development, and secondary metabolism; yet it was up-regulated significantly with the infection of Fusarium oxysporum in root, suggesting that it might be involved in defense against many diseases including root rot in P. notoginseng.

关 键 词：三七 病程相关蛋白 基因克隆 表达模式 防御反应 

分 类 号：R931[医药卫生—生药学]