Antimicrobial agents are essentially important in reducing the global burden of infectious diseases. However, emergence and dissemination of multidrug resistant (MDR) strain in pathogenic bacteria have become a significant public health threat as there are fewer, or even sometimes no, efective antimicrobial agents available for the infection caused by pathogenic bacteria. In the present study aimed to investigate in vitro antibacterial and antifungal activity of Hibiscus micranthus leaf and best callus extract. Pathogenic microbes including bacteria as Escherichia coli, Streptococcus pyogenes, Psudomonus aurginosa, Salmonella typhi specie of bacteria was spread on Nutrient agar plates and fungi as Aspergillus fumigates, Cryptococcus neoformans, Sporothrix schenckii and Phialophora verrucosa. it can be concluded that the ethanolic extract of both the mother plant and its callus showed generally the great antimicrobial activities against 4 types of bacteria and 4 fungus species tested. It was observed that the callus extract was potent antimicrobial against all tested organisms than mother plan extract. The antimicrobial activity of callus extract due to the presence of higher content of phenol and flavonoids.