马铃薯黑痣病多重实时定量PCR检测方法的建立  

Establishment of Multiple Real-time Quantification PCR Detection Method for Potato Black Scurf

作　　者：陈恩发 卢扬 彭慧元 雷尊国 邓宽平 

机构地区：[1]贵州省生物技术研究所,贵州贵阳550006

出　　处：《贵州农业科学》2014年第11期136-139,143,共5页Guizhou Agricultural Sciences

基　　金：贵州省科技厅农业攻关项目"马铃薯微型薯环保型种衣剂研究与应用"[黔科合NY字(2010)3012];贵州省农业科学院专项资金项目"贵州马铃薯晚疫病数字防控技术研究与应用"[黔农科院院专项(2013)007];贵州省国际科技合作项目"马铃薯晚疫病抗性材料引进与田间评价"[黔科合外G字(2012)7026]

摘　　要：为建立快速、准确、可靠的马铃薯黑痣病鉴定和检测方法,以黑痣病病原菌立枯丝核菌核糖体基因簇间的保守序列—转录间隔区（Internal transcribed spacer,ITS）为靶区域,同时引入马铃薯细胞色素氧化酶（cytochrome oxidase gene,Cox）作内参,应用多重实时定量PCR技术,建立马铃薯黑痣病病原菌TaqMan RFQ-PCR多重快速检测方法。结果表明：RFQ-PCR检测体系的引物-探针具有高度的特异性,灵敏度比普通PCR方法高10～100倍;内参基因的引入还可有效避免假阴性的出现。方法特异性强、灵敏度高、简单、快速、可靠。The TaqMan RFQ-PCR multiple real-time quantification detection method was established to detect conserved sequence-Internal transcribed spacer（ITS）and Cox gene of Rhizoctonia solani Kn rDNA gene cluster of potato black scurf pathogenic bacteria simultaneously.The results showed that the new RFQ-PCR multiple real-time quantification detection method with advantages of strong specificity,high sensitivity,simplicity,rapidity and reliability can be used in detection of Rhizoctonia solani of potato black scurf because its primer-probe is of higher specificity and higher sensitivity,and introduction of reference gene can avoid appearance of false negative effectively.

关 键 词：马铃薯黑痣病 检测 多重实时定量PCR 检测方法 

分 类 号：S532[农业科学—作物学]