Real-Time PCR and Its Application in Plant Disease Diagnostics

Abstract

Real-time PCR is currently considered as the gold standard method for detection of plant pathogens. This technique allows the monitoring of the reaction during the amplification process by the use of a fluorescent signal that increases proportionally to the number of amplicons generated and to the number of targets present in the sample. Real-time PCR makes possible an accurate, reliable and high-throughput quantification of target pathogen DNA in various environmental samples, including hosts tissues, soil, water and air, thus opening new research opportunities for the study of diagnosis, inoculum threshold levels, and epidemiology and host pathogen interactions. Real-time PCR has versatile practical application in diagnostics of plant disease. With Real-time PCR, it is possible not only to identify and detect the presence or absence of the target pathogen, but it is also possible to quantify the amount present in the sample allowing the quantitative assessment of the number of the pathogen in the sample. Enumerating the pathogen upon detection is crucial to estimate the potential risks with respect to diseases development and provides a useful basis for diseases management decisions. Determination of the viability of a pathogen, detection of multiplexing and monitoring fungicide resistance in pathogens are other major application areas. Generally, real-time PCR technologies open increasing opportunities and a significant role in better understanding of the dynamics of plant pathogenic microorganism and, thereby allow better management of the diseases.

Key words:  Diagnostics, high-throughput, Pathogen, Quantification, Real-time PCR