文档介绍：中间偃麦草NPR1同源基因TiNH1的分离和特性分析
唐益苗,张增艳,辛志勇
(中国农业科学院作物科学研究所,农作物基因资源与基因改良国家重大科学工程,农业部作物遗传育种重点开放实验室,北京 100081)
摘要:【目的】NPR1是调控植物抗病反应的一个关键基因。对中间偃麦草NPR1同源基因TiNH1进行分离和特性分析。【方法】利用RT-PCR和RACE(rapid amplification of cDNA ends)技术获得TiNH1全长cDNA序列,利用Northern和Southern分别研究TiNH1表达特性及其在中间偃麦草基因组中存在形式。【结果】获得了该基因cDNA序列,命名为TiNH1。其编码蛋白TiNH1的氨基酸序列分别与水稻、烟草和拟南芥的NPR1同源性为80%、54%和46%。Northern杂交分析结果表明:TiNH1基因在正常情况下有微量表达,在小麦白粉病菌和纹枯病菌诱导下,该基因表达水平得到提高。Southern杂交分析结果表明;该基因以单拷贝形式存在于中间偃麦草基因组中。【结论】中间偃麦草NPR1同源基因TiNH1编码由580个氨基酸组成的蛋白质TiNH,具有已知NPR1蛋白保守的结构域和功能氨基酸,可能参与寄主对小麦白粉病菌和纹枯病菌的防御反应。
关键词:中间偃麦草;NPR1同源基因;白粉病菌;纹枯病菌;抗性
Isolation and Characterization of NPR1 Homolog Gene TiNH1 in Thinopyrum intermedium
TANG Yi-miao, ZHANG Zeng-yan, XIN Zhi-yong
(National Key Facility for Crop Gene Resources and ic Improvement, Key Laboratory of Crop ics and Breeding, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081)
Abstract: 【Objective】NPR1 is a key regulator in plant defense response. The NPR1 homolog gene TiNH1, named as TiNH1 was isolated from Th. intermedium and characterized. 【Method】The full-length cDNA of TiNH1 gene was isolated by RT-PCR and RACE (rapid amplification of cDNA ends ) methods. The expression pattern of TiNH1was studied by Northern blot and its copy numbers in Th. intermedium genome was clarified by Southern blot. 【Result】The full-length cDNA of TiNH1 was obtained from Th. int