As medicinal plants are generally collected from the wild flora, it is of fundamental importance to identify the optimum growth conditions for increasing the yield and quality potential of ex-situ cultivated plants. Effects of different soil water content [20, 40, 60 and 80 % of water holding capacity (WHC)] on growth, physiology and biochemical profile of Achillea millefolium group-related cv. Proa were investigated in the present study. Alterations in drug herbage, chlorophyll content, quantum efficiency of photosystem II, essential oil rate and composition, proline content and lipid peroxidation, antioxidant activity and flavonoid content of the plants under different plant and soil water status were determined. Decrease in soil water status under 40 % of WHC resulted in degradation of chlorophyll pigments, reduction in photochemical efficiency and lower dry matter production. The essential oil (EO) rate of cv. Proa was higher under limited and excessive water content of soil whereas the main component of EO, chamazulene, was negatively correlated with WHC of soil. Our results suggest a general trend of inverse correlation of antioxidant capacity and flavonoid content with essential oil rate in contrast to the positive correlation of these parameters with chamazulene content. The data obtained may serve as a basis of programming the optimum soil moisture conditions for successful cultivation of cv.Proa.