Pathogens decrease flax yield and reduce product quality. The development of molecular markers is necessary for fungus identification and application of proper defense actions. In our work, we present the method based on deep sequencing for studying the genetic diversity of fungal pathogens of flax. ITS regions of rRNA genes and regions of genes encoding beta-tubulin (tub2), translation elongation factor 1-alpha (tef1), and RNA polymerase II subunits (RPB1 and RPB2) were studied using deep sequencing in 100 pathogen strains of Fusarium , Colletotrichum , Melampsora , Aureobasidium , and Septoria . Our method can be used for the characterization of genetic diversity of pathogens and determination of DNA sequences that are the most suitable for identification of fungus using molecular markers.