In the present investigation, the root-colonizing bacteria were isolated on different media like Pikovskaya¶s Agar media for Phospahte solublizer, King¶s B media for Fluoresceent Pseudomonas and Actinomycetes Agar media for Actinomycetes and screened using Dual culture analysis on N.A. medium to select antagonistic bacteria from a random group of bacterial isolates and to compare with selected strains for their fungal growth inhibition capabilities. In this way 15 bacterial strains displaying a wide antifungal spectrum were selected as the most interesting bacteria to test for biological control of green mold disease caused by Trichoderma harzianum. A total of 17 strains of Actionmomycetes were isolated from different soil samples on the basis of different parameters of colony characteristic like colony shape, size, surface, margin and color on the actinomycetes agar medium. Most of these isolates were from Rose and Wheat soil. Biochemical characterization including Gram-staining test, IAA test, Indole-production test, MRVP test and Antibiotic assay of different bacterial strains were performed successfully. Molecular characterization followed by PCR amplification based 16S rDNA of antagonistic bacterial strains was done. The bacterial strains such as PKB-1, PPB-4, PAM-5, PPB-3, PKB-19, PAM-11, PAM-12, PKB-9, and PKB-18 showed clear DNA bands. The bacterial strains which were successfully amplified by PCR were PKB-1, PKB-18, PPB-3, PPB-4, and PAM-11 along with the ladder