PCR Based Detection Of Entomopathogenic Fungus Metarhizium Anisopliae In Host Organisms

PCR based detection and identification of the entomopathogenic fungus Metarhizium anisopliae was conducted with specific primers F3 (5'-GGGTATATGAGAGGGAGGGC-3') and B3 (5'- GGTTCCTGGTCGGGACTT-3') which amplify a fragment of gene in the IGS (Intergenic spacer) region of rRNA (Ribosomal RNA) of M. anisopliae. The PCR amplification of IGS sequences yielded a unique fragment of 226-bp for all the four strains of M. anisopliae (M4, M16, M34 and M43). The results proved that the primers F3 and B3 were highly specific for M. anisopliae. PCR based detection M. anisopliae within host insects as Mealworm beetle (Tenebrio molitor) in the laboratory and cockchafer (Melolontha spp) in the field by using specific primers was applied. The PCR method could be a simple, rapid method to detect M. anisopliae within host insects just 8 days after infection. This study also showed that M. anisopliae exists in the soils in Fels??rs-Koveskutpuszta region in Hungary. In fact, the results proved that DNA extracted from infected insects in laboratory and field could be used to identify the presence of the entomopathogen fungus M. anisopliae by using specific primers. Our study demonstrates an alternative approach for typing M. anisopliae strains within infected insects and reduces the need for time-consuming morphological and physiological tests.