A study was conducted in the Department of Plant Protection, College of Agriculture, University of Anbar, aiming to identify the major fungi causing root and crown rot disease in strawberries through isolation and diagnosis using both morphological and molecular methods. the results of isolation and diagnosis from the affected plant parts revealed the prevalence of root and crown rot disease in all sampled areas exhibiting symptoms and signs of the disease. Seventeen isolates were obtained belonging to the Fusarium species, eight isolates belonged to the fungus Rhizoctonia solani, and seven isolates belonged to the spp. Neopestalotiapsis The fungal species were identified based on their taxonomic characteristics. DNA extraction was successfully performed on eleven isolated fungi from the roots and crowns of strawberry plants in different isolation areas. Partial diagnosis was conducted using PCR technique targeting the intergenic spacer region using primers (ITS1-ITS4) to obtain amplicons of 650 base pairs (bp) in size. The amplicons were then separated on 2% agarose gel, and the nucleotide base sequences of these isolates were determined and deposited in the gene bank.