摘要
筛选到应用于多主棒孢实时荧光定量PCR检测的特异性引物CIR5/CIF5,并建立了土壤中多主棒孢的实时荧光定量PCR检测技术。引物CIR5/CIF5能够从多主棒孢基因组DNA中特异性扩增出一条259 bp的片段,常规PCR检测中灵敏度可达10 fg/μL。利用实时荧光定量PCR检测模拟带菌土壤中的带菌量,结果证明该技术对土壤中多主棒孢的检测下限为1个孢子/g土壤,可以快速、准确地定量土壤中病原菌的孢子数目,为栽培前土壤中病原菌的监测预报提供有效的技术手段。
In this research,a specific primer pair CIR5 /CIF5 for C. cassiicola was developed,and Real-time PCR based specific detection method of this pathogen in soil was established. The primer pair gave a single amplification of 259 bp from C. cassiicola and could be distinguished from other soil-borne pathogen strains. The detection limit was 10 fg /μL in conventional PCR. The pathogen in soil could be detected by Real-time quantitative PCR,and the sensitivity was down to C. cassiicola DNA of 1 conidia / g. This Real-time quantitative PCR method could assist in the implementation of quarantine measures for prevention and control of Corynespora spot leaf.
出处
《华北农学报》
CSCD
北大核心
2014年第2期71-74,共4页
Acta Agriculturae Boreali-Sinica
基金
大宗蔬菜产业技术体系建设专项(CARS-25)
关键词
多主棒孢
实时荧光定量PCR
土壤检测
Corynespora cassiicola
Real-time quantitative PCR
Soil detection
