CAJ | 학술논문

为建立土壤中黑胫病菌的SYBR Green I实时荧光定量PCR快速检测方法，依据疫霉菌特异性parA1基因序列差异位点，设计1对特异性引物。通过构建含有目的片段的重组质粒，建立标准曲线，以SYBR Green I荧光染料法对烟草黑胫病菌进行荧光定量PCR检测，检测灵敏度达1×102个／g土壤，建立了基于SYBR Green I荧光染料法的荧光定量PCR的快速定量检测体系，并对云南、四川烟草主要栽培区土壤进行检测。表明该体系可以对烟草不同时期的田间土壤黑胫病菌的数量进行动态监测，可用于烟草黑胫病的预测和防治。
위건립토양중흑경병균적SYBR Green I실시형광정량PCR쾌속검측방법，의거역매균특이성parA1기인서렬차이위점，설계1대특이성인물。통과구건함유목적편단적중조질립，건립표준곡선，이SYBR Green I형광염료법대연초흑경병균진행형광정량PCR검측，검측령민도체1×102개／g토양，건립료기우SYBR Green I형광염료법적형광정량PCR적쾌속정량검측체계，병대운남、사천연초주요재배구토양진행검측。표명해체계가이대연초불동시기적전간토양흑경병균적수량진행동태감측，가용우연초흑경병적예측화방치。
The aim of this study was to establish the SYBR Green I real-time fluorescence quantitative PCR assay for rapid detection of Phytophthora nicotianae in soil.According to the sequence of the parA1 gene, the specific pair of primers were designed.By constructing a recombinant plasmid containing the desired frag-ment and establishing the standard curve,the real-time qPCR with SYBR Green I dye fluorescence was used to detect Phytophthora nicotianae in soil.The sensitivity of detection was Up to 1×102 spores per gram of soil.A rapid quantitative detection system of qPCR based on SYBR Green I fluorescence dye method was established, for the detection of the soil of the main tobacco cultivation area soil in Yunnan and Sichuan.These results indi-cated that the amount of Phytophthora nicotianae in different tobacco growth periods of field soil can be dynam-ically monitored by this system,and could be used in predicting the occurrence of tobacco black shank.