Development and application of multiplex PCR for identification of three fungal species of the genus Parastagonospora widespread in Russia

A multiplex PCR system has been developed and validated for the specific detection of three prevalent in Russia cereal pathogens, belonging to the genus of Parastagonospora: P. nodorum, P. avenae, and P. pseudonodorum. The target region selected was a fragment of the YPEL (Yippee-like protein) gene, which exhibits sufficient interspecies polymorphism for designing highly specific primers. Primer specificity was confirmed in silico and in vitro using a collection of reference isolates. The developed primers functioned effectively in a multiplex format, enabling species-specific detection. The system demonstrated high sensitivity (detection limit: 1 pg of genomic DNA per reaction). This assay can be utilized for phytosanitary monitoring and research on the epidemiology of Septoria diseases in cereal crops.