CAJ | 학술논문

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。
사선도응용우다주봉포실시형광정량PCR검측적특이성인물CIR5/CIF5,병건립료토양중다주봉포적실시형광정량PCR검측기술。인물CIR5/CIF5능구종다주봉포기인조DNA중특이성확증출일조259 bp적편단,상규PCR검측중령민도가체10 fg/μL。이용실시형광정량PCR검측모의대균토양중적대균량,결과증명해기술대토양중다주봉포적검측하한위1개포자/g토양,가이쾌속、준학지정량토양중병원균적포자수목,위재배전토양중병원균적감측예보제공유효적기술수단。
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 amplifi-cation 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.