A rapid visualization method for the detection of the tea plant pathogen Colletotrichum camelliae by qPCR
[Objective]This study was conducted in order to establish a real-time quantitative PCR method for the rapid detection of Colletotrichum camelliae.[Method]Using C.camelliae as the pathogen of tea anthracnose as detection target,the specific primers were designed according to ApMat gene fragments,and the qPCR detection technique system was established.The specificity of the primers was verified,the sensitivity of the test reaction was obtained and the standard curve was drawn.Finally the detection effect was tested by collecting and detecting the diseased leaves in the field were collected.[Result]The results showed that the primers of CCF/CCR had the best specificity and sensitivity,for detecting C.camellae and had the highest amplification efficiency.The sensitivity of C.camelliae was 10 pg/μL,and the linear relationship between the logarithm of DNA concentration(X)and CT value(Y)was Y=-3.5296x+36.938(R2=0.9957,E=92.01).The specific peak Tm of the dissolution curve of C.camelliae was(85.5±0.5)℃.The pathogen C.camelliae was detected successfully in the diseased leaves from both artificially and naturally infection.[Conclusion]The established qPCR reaction system can be used to detect C.camelliae,and can also be applied to the early diagnosis of diseased leaves in the field.
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