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In Pakistan, strawberry (Fragaria ananassa Duch.) is emerging as an economically important commercial fruit crop. Gray mold or Botrytis fruit rot of strawberry is widely distributed and economically important disease infecting the strawberry fruit at both pre- and post-harvest stages. To assess the percent disease prevalence and incidence, a comprehensive survey was carried out in 8 strawberry growing districts of Punjab viz. Rawalpindi, Sargodha, Gujranwala, Sialkot, Narowal, Sheikhupura, Lahore and Multan during crop seasons of 2014-15 and 2015-16. Disease prevalence and incidence ranges were found to be 87.5%-100% and 29.3%-48.13%, respectively in the studied areas. The highest disease incidence was recorded in Narowal district (48.13%) followed by Gujranwala (46.36%) and Sialkot (45.97%), whereas the lowest was seen in Rawalpindi (29.3%). Characterization of 61 isolates recovered from the study areas revealed that pure colonies of the fungus were cottony white to grayish in color. Fungus produced obovoid or ellipsoid conidia with 11-16 × 5-8 µm size and conidiophores ranged from 735 to 2805 µm along with production of dark brown to black sclerotia measured 2-7 mm. A total of 8 representative and highly virulent isolates one from each district were subjected to DNA amplified using inter transcribed spacer regions ITS (ITS1, 5.8S, ITS2) and submitted to GenBank database of NCBI with accession numbers MF959755 to MF959762, respectively. Sequences revealed 99 to 100% genetic similarity with Botrytis cinerea reference isolates. Phylogenetic analysis was done by constructing evolutionary tree with previously published sequences of B. cinerea in NCBI from different parts of world. Analysis of evolutionary tree highlighted three diverse sub trees. This is the first ever detailed study of its kind to know the status of the disease along with its distribution and characterization on this relatively new crop in Punjab, Pakistan.