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The present studies were undertaken to evaluate the effect of nitrogen source on the mycelium growth of Pleurotus pulmonarius and to investigate the antimicrobial activities of ethanolic extracts of the three mushrooms (Pleurotus pulmonarius. Pleurotus florida and Schizophyllum commune). Assessment of different nitrogen sources revealed that maximum mycelial growth of P. pulmonarius was recorded in potassium nitrate followed by diammonium hydrogen phosphate, sodium nitrite and ammonium nitrate. The ethanolic extracts of three mushroom observed to inhibit the growth of plant pathogenic bacteria such as Xanthomonas axonopodis pv. punicae, Bacillus pumilus and Pseudomonas syringe. The growth of plant pathogenic fungi like Aspergillus niger, Aspergillus flavus, Alternaria solani, Colletotricum graminicola and Fusarium oxysporum, were also inhibited in the presence of the extracts of three mushroom. It is also evident from the results that the gram-positive bacterium (Bacillus pumilus) was more sensitive than gram-negative bacteria (Pseudomonas syringe and Xanthomonas axonopodis pv. punicae) to the examined macrofungi extracts.