Apple (Malus domestica Borkh L., Family Rosaceae) is among the most popular fruits worldwide. Apple fruits are attacked by several fungal pathogens both pre- and post-harvest. The post-harvest fruit rot of apples is one of the most threatening dis-eases in apple cultivation. In the current study, we aimed to isolate, characterize, and identify the fungal phytopathogens associated with postharvest rot of apple fruits. Herein, we isolated eight genera from rotted Anna apple fruits during the 2018 season from orchards, wholesale, and retail markets as well as refrigerators. Then, the dif-ferent fungal pathogens were identified according to their cultural and microscopic characteristics as Penicillium expansum, Alternaria alternate, Botrytis cinerea, As-pergillus niger, Mucor sp., Rhizopus stolonifer, Stemphyllum sp., and Trichothecium roseum. In this study, investigations were made into the pathogenicity tests of the highest-frequency fungi. Pathogenicity tests on apples (Anna cv.) revealed that B. cinerea was the highest virulent fungus. Additionally, a simple linear regression (SLR) analysis was carried out to better understand the relationship between the di-ameter of the infected area caused by post-harvest pathogens and storage time. Therefore, B. cinerea was identified by molecular testing using internal transcribed spacer (ITS) sequencing and phylogenetic analyses. Postharvest gray mold, caused by Botrytis cinerea, is one of the most important diseases and seriously affects apple fruit quality, resulting in huge economic losses. Proper measures should be adopted to protect apple fruits from fungal decay