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Ammi visnaga (L.) Lam is a valuable herbal plant that is frequently collected for medicinal purposes. This study is conducted to evaluate the antimicrobial potential of the in-vitro grown microshoots and callus cultures of this plant against selected strains of bacteria and fungi. Shoot multiplication was obtained in MS medium containing 0.5 mg/L BA + 0.1 mg/L NAA, while callus multiplication was performed on MS medium containing 1.0 mg/L BA + 2.0 mg/L 2,4-D under light conditions. The aqueous and methanolic extracts were prepared from both culture types in addition to in-vivo grown plant material to experiment their antimicrobial activities. Generally, the in-vitro extracts of the microshoots and the callus cultures acted best against bacteria compared to the field plant extract. Moreover, the methanolic extracts were generally found to exhibit far better results and resistance against the tested microbes than the aqueous extracts. C. albicans was the most sensitive species to the microshoots extracts followed by gram-positive bacteria at MIC values of (3.125, 6.25) mg/mL. Meanwhile, E. coli bacteria were most sensitive to the microshoot extracts as they were completely inhibited at MIC value of (0.78 mg/mL). Moreover, the results of the callus extracts showed that C. albicans was the most sensitive at MIC values of (1.56 and 3.125) mg/mL, followed by gram-positive bacteria. On the other hand, gram-negative bacteria were the most resistant microbes to all experimented extract types in this study. [ABSTRACT FROM AUTHOR]