马铃薯黑胫病菌的分离、纯化及PCR检测  

Isolation, Purification and PCR Detection of Pectobacterium atroseptica from Potato

作　　者：林燕文[1] 陈妙英[1] 毛露甜[1] 陈兆贵[1] 

机构地区：[1]惠州学院生命科学系,广东惠州516007

出　　处：《中国马铃薯》2016年第3期169-174,共6页Chinese Potato Journal

基　　金：广东省科技计划项目"冬种马铃薯健康种薯质量检测和控制技术体系研究(2011B020303010)";广东省科技计划项目"南方冬种马铃薯健康种薯繁育及高效栽植技术推广应用(2013B090200023)"

摘　　要：由Pectobacterium atroseptica引起的马铃薯黑胫病是影响马铃薯生产的重要种薯传播细菌病害,从种薯发芽到生长后期均可发病,严重影响马铃薯的产量和品质。为构建马铃薯黑胫病菌的分离、鉴定技术体系,从广东省惠州市惠东县大水坑马铃薯基地采集了疑似感染马铃薯黑胫病植株,对其进行病原菌分离和纯化,得到7株病菌,利用引物Eca 1 g/Eca 2 g对其进行PCR扩增、将PCR产物测序并将所测定序列递交Gen Bank数据库,使用Blast软件进行比对确定目标菌株。结果表明,其中一株菌株能扩增出长度大约688 bp的特异性条带,其DNA序列符合Eca 3762序列,鉴定为马铃薯黑胫病菌。此方法可以成功地从发病植株中分离出黑胫病菌,成本较低廉,为广东省建立马铃薯种薯质量检测和控制体系提供技术支撑。Potato black leg, caused by Pectobacterium atroseptica, is a major bacterial disease influencing potatoproduction. It is spread by seed potato, and may occur in any stage from germination to late growth and seriously affectthe yield and quality of potatoes. To establish a technical system which can isolate and identify P. atroseptica frompotatoes, the samples suspected of being infected by the pathogen were collected from Dashuikeng of Huidong Countyin Huizhou City. Seven suspected bacterial objects were isolated and further purified. Their specific DNA fragmentswere amplified with primer Eca 1 g/Eca 2 g by PCR, and the fragments were sequenced and aligned with GenBankdatabase. Finally, the bacterial objects were compared and determined by using Blast software. The results showed that aspecified fragment about 688 bp was amplified in one of the objects, and its sequence matched that of Eca 3762. So it canbe identified as P. atroseptica. This method could successfully isolate P. atroseptica from diseased potato plants with low cost,so it may provide a technical support for the seed potato quality detection and control system of Guangdong Province.

关 键 词：马铃薯 黑胫病菌 马铃薯黑胫病 CVP培养基 PCR检测 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]