In the present study, the alkaloid composition of the aerial parts of Genista vuralii A. Duran & H. Dural (Fabaceae) was investigated by capillary GC-MS. Ten quinolizidine alkaloids were identified by capillary GC-MS, namely, N-methylcytisine, cytisine, tetrahydrorhombifoline, 17-oxosparteine, 5,6-dehydrolupanine, lupanine, 17-oxolupanine, anagyrine, baptifoline, and 13 alpha-tigloyloxylupanine. Among them, anagyrine (93.04%) was the most abundant alkaloid. Furthermore, antibacterial and antifungal activities of the alkaloid extract of G. vuralii were tested against standard strains of bacteria (Escherichia coli, Pseudomonos aeruginosa, Bacillus subtilis, Staphylococcus aureus) as well as fungi (Candida albicans, Candida krusei). The alkaloid extract of G. vuralii presented good activity against S. aureus, B. subtilis, and C krusei, with minimum inhibitory concentrations (MIC) of 62.5 mu g/mL. The remaining MIC values were found to range between 125 and 500 mu g/mL. To the best of our knowledge, the current work is the first to report the alkaloid profile and antimicrobial activity of G. vuralii L. growing in Turkey.