Spinach (Spinacia oleraceae L.) is an edible annual crop, grown worldwide for its high nutritious value. In the recent past, spinach is found to be threatened by various fungal leaf spot diseases which are among the highest cause of yield loss in spinach. In the present study, a field survey was conducted in August-September 2017 and 2018 in the fields of Okara, Depalpur city, vegetable area of University of the Punjab, Lahore, and Experimental area of Institute of Agricultural Sciences, University of the Punjab, Lahore where more than 55-60% plants of S. oleraceae were infected with leaf spots disease. Isolation and identification of fungal pathogen by morphological and molecular techniques confirmed that Aspergillus aculeatus Lizuka. was the leaf spot disease-causing agent of S. oleraceae. The pathogenicity trails confirmed A. aculeatusas the virulent pathogen of spinach. This study concerns the first report of A. aculeatus as a pathogen of S. oleraceae which necessitates the quick development of management tools.