AbstractUnderstanding of the physiological role of mammalian hormone—androstenedione (AN)—in plants is scant and the mechanisms of its action at a cellular level are practically unknown. The aim of this study was to investigate the physicochemical and biochemical background of AN activity in winter wheat exposed to low temperature. Cold periods are important in the lifecycle of this species as they induce frost resistance and further generative development. Wheat seedlings (control and AN-supplemented) were acclimated 2 weeks in cold and then exposed to frost (−12 °C). AN supplementation reduced frost damages by 30%. Moreover, AN also accelerated generative development of wheat. The AN-induced changes in redox homeostasis seemed to be important for processes of acclimation to low temperature and generative induction. AN influenced hormonal balance in wheat and stimulated accumulation among other gibberellins and cytokinins. For example, in aerial part of plants, the content of GA₃was increased by AN in 12 days of cold by about 30%, whereas the content ofcis-zeatin was increased by 65%. AN was absorbed into plant membranes (Langmuir bath studies). The membrane absorption of AN increased the distance between lipid molecules and this may be an important step in the AN-induced enhancement of frost resistance. AN interaction with lipid membranes showed similarity to the interactions of some known regulators stimulating flowering in plants, and thus it may also underlie the acceleration of wheat development. Androstenedione was naturally present in wheat leaves (5–21 pg g⁻¹ FW).