摘要
甘蓝枯萎病菌生理小种传统鉴定方法费时费力,不能满足生产的要求,因此需要建立一种快速、可靠的分子检测技术。本研究在甘蓝枯萎病菌1号和2号生理小种基因组测序的基础上,通过比较基因组学方法筛选1、2号生理小种各自的特异基因片段并设计引物,并分别以10个甘蓝枯萎病菌1号生理小种菌株、2个2号生理小种菌株、7个尖孢镰刀菌其他专化型菌株及4个外围菌株DNA为模板进行常规PCR扩增,筛选出甘蓝枯萎病菌1号和2号生理小种特异性引物,同时引入尖孢镰刀菌通用引物W106R/W106S,建立起一步三重PCR检测甘蓝枯萎病菌1、2号生理小种的分子检测技术。结果表明,该分子检测技术实现了在一次PCR反应中快速、准确地同步检测出甘蓝枯萎病菌DNA、罹病甘蓝组织和土壤中的甘蓝枯萎病菌1号和2号生理小种,对检测甘蓝植株是否感染枯萎菌及甘蓝种植区土壤是否受到枯萎菌的污染有实用价值。
The traditional race identification of Fusarium oxysporum f. sp. conglutinans （FOC） is laborious and time-consuming, and can＇t meet the requirements of practical production. Development of a molecular method for rapid detection and identification of race 1 and race 2 is therefore eagerly needed. In this study, specific primers of race ] and race 2 were screened on the basis of comparative genome analysis of these two races and other formae specials of F. oxysporum. In addition, DNAs of ten FOC race 1 isolates, two FOC race 2 isolates, seven isolates of other races of FOC and four non-FOC isolates were used as templates for further evaluation of the specificity of two primers specific to FOC race 1 and race 2, respectively. Moreover, another primer set （W106R/F]06S） was used to develop a triple PCR method for the detection and identification of FOC race 1 and race 2 in one PCR reaction. It was confirmed that the triple PCR method can not only detect and identify FOC race 1 and race 2 from pure DNA samples but also from the infected plant tissues and soil samples, and therefore has application value to detect FOC races from the infected plant tissues and soils.
出处
《植物病理学报》
CAS
CSCD
北大核心
2014年第6期586-594,共9页
Acta Phytopathologica Sinica
基金
国家自然科学基金项目(31272003)
公益性行业(农业)科研专项(200903049-04)
国家科技支撑计划项目(2012BAD19B06)
国家大宗蔬菜产业技术体系建设项目(CARS-5-B-01)
农业部园艺作物生物学与种质创制重点实验室
中国农业科学院科技创新工程
