Development of multiplex PCR assay for simultaneous detection of five cucumber pathogens based on comparative genomics

The aim of this study was to develop a multiplex PCR assay for rapid, reliable and simultaneous detection of five important cucumber pathogens: Fusarium oxysporum, F. solani, Sclerotinia sclerotiorum, Pythium aphanidermatum and Botrytis cinerea. One common forward primer and species-specific reverse primers were designed based on comparative genomics. The specificity of the developed multiplex PCR was confirmed using target pathogens and other closely related pathogens. Furthermore, this method accurately identified the presence of pathogens among cucumber field samples. Consequently, we established a multiplex PCR method which could provide reliable and informative identification of five cucumber pathogens in the tested environmental samples.