Used as an antibiotic to treat bacterial infections, particularly tuberculosis caused by Mycobacterium tuberculosis. Effective against a range of gram-negative bacteria (e.g., Yersinia pestis causing plague, Brucella spp.) and some gram-positive bacteria. Often used in combination with other antibiotics to prevent resistance. Commonly applied in medical treatment for respiratory or urinary tract infections resistant to other antibiotics, veterinary medicine, and laboratory research for selecting genetically modified cells using streptomycin resistance genes.

In agriculture, used as a bactericide for plants to control bacterial diseases in vegetables and fruits, such as soft rot in potatoes or leaf spot in crops, typically applied as a foliar spray solution.