In the present investigation, antimicrobial activity and phytochemical screening was evaluated for the three newly reported macro fungal species from Rajouri District, J&K, India. The hydro-alcoholic extracts of the dried mushroom samples viz. Krombholziella oxydabilis (A), Daldenia concentrica (B) and Innotus radiate (C) at 200 mg/ml were evaluated against bacterial and fungal organisms viz. Pseudomonas aeruginosa, Bacillus licheniformis, Micrococcus luteus, Aspergillus niger and Cladosporium herbarum. The results showed that mushroom species viz. Krombholziella oxydabilis and Innotus radiate can be utilized as a suitable antimicrobial agent while mushroom species, Daldenia concentrica doesn’t possessed antimicrobial activity against any of the fungal and bacterial strains studied.