BACKGROUND The aim of this experiment was to determine the essential oil, flavonoid and phenolic acid contents as well as yields of raw material collected from different morphological parts of Greek oregano in five consecutive developmental stages. Antibacterial and antifungal properties of methanol and hexane extracts from the aerial parts of Greek oregano were assessed as well. RESULTS The findings showed that Greek oregano grown in the cool climate of central Europe had a considerable variation in the essential oil yield and chemical composition related to the harvesting stage. Forty volatile compounds were detected in essential oils, with the highest amount of carvacrol, thymol, p‐cymene and γ‐terpinene, constituting in total from 90.72 to 92.9%. CONCLUSION Different morphological parts of the plants contained different amounts of essential oils, with dominant amounts in the flowers. The essential oil content systematically increased, reaching its maximum in the full blooming period. In that phase, the plants produced the highest yields of aboveground parts and provided approximately 190 L ha−1 of essential oils. Mainly owing to its content of phenolic compounds, the Greek oregano raw material exhibited relatively high antioxidant activity. The data obtained clearly showed the inhibitory activity of methanol and hexane extracts against pathogenic bacteria and fungi. © 2019 Society of Chemical Industry