Present study is carried out to evaluate the solvent extracts of six day old culture of Penciillium steckii strain MF 1 grown in modified PDA medium. The mass culture of P. steckii grown in two different modified medium were treated with different solvents like ethanol, ethyl acetate and their combination with water. The ethyl acetate and ethanol fractions were tested against the dieback and verticillium pathogenic fungi in agar plate culture. Crude i.e. ethyl acetate fraction and fractions eluted with ethyl acetate had shown positive activity against the dieback pathogenic fungi where as three water fractions were exhibited the inhibitory activity against verticillium pathogenic fungi. UV spectrum at 284 nm of this fraction showed probability of being nystatin like antifungal compound. The present work indicates that our fungal strain P. steckii has possibility to produce antifungal compound against dieback and verticillium pathogenic fungi of rose. This antifungal compound may be of nystatin or its derivative that need further experimental work on extraction and purification of active compound.