人工microRNAs干扰At3A06基因增强了拟南芥对菌核病的敏感性  

Increased susceptibility of Arabidopsis to Sclerotinia sclerotiorum by artificial microRNAs silenced At3A06 gene

作　　者：黄军艳[1] 许李明[1,2] 张学江[1] 刘胜毅[1] 

机构地区：[1]农业部油料作物生物学重点开放实验室中国农业科学院油料作物研究所,湖北武汉430062 [2]中南民族大学生命科学院,湖北武汉430074

出　　处：《中国油料作物学报》2010年第3期345-348,共4页Chinese Journal of Oil Crop Sciences

基　　金：中国农业部油料作物生物学重点开放实验室开放课题项目(2007011);国家自然科学基金项目(30900934);公益性行业(农业)科研专项项目(3-21)

摘　　要：基于本研究室自主开发的油菜防御cDNA芯片分析结果,油菜基因Bn3A06受菌核病诱导后特异上调表达。序列比对发现,该基因与拟南芥基因At3A06高度同源,后者编码289个氨基酸,功能未知。本研究用花序浸染法将At3A06基因的人工微RNA(amiRNA)干扰载体(该载体包含一个除草剂草胺磷抗性基因)转化拟南芥。经草胺磷筛选获得3株T1代转基因植株,PCR检测均为阳性;荧光定量PCR检测表明,与野生型对照相比,3个转基因植株的At3A06基因的表达量均显著降低;对其T2代进行菌核病抗性鉴定,结果显示转基因株系对菌核病的敏感性均显著增强(P<0.05)。推测At3A06基因可能参与了植物的防御反应,与植株对核盘菌抗性相关。Bn3A06 gene of Brassica napus L.was obviously up-regulated in gene microarray-profiling（cDNA microarray） experiments in which plants were infected by Sclerotinia sclerotiorum.The homologous gene of Bn3A06 from Arabidopsis thaliana named At3A06 encodes 289 amino acids whose function is unknown.Then the At3A06 function was studied using an artificial microRNA（amiRNA） expression vector targeting At3A06 after Agrobacterium-mediated floral dip transformed into A.thaliana.The vector harbors a bar gene conferring herbicide-resistance.There were three T1 transgenic plants obtained by herbicide screening and PCR identification.The At3A06 gene expression,detected by real-time quantitative PCR,was significantly inhibited in all transgenic plants.When inoculated S.sclerotiorum of the detached leaves of T2 plants,higher susceptibility to the pathogens was found than that of the non-transgenic plants,which suggested that At3A06 gene may play an important role in resistance to S.sclerotiorum.

关 键 词：拟南芥 核盘菌 防御反应 人工microRNAs 荧光定量PCR 

分 类 号：S565.401[农业科学—作物学] Q344.13[生物学—遗传学]