Polyethylene glycol (PEG 6000)-induced water deficit causes physiological as well as biochemical changes in plants. The present study reports on the results of such changes in hydroponically grown tomato plants (Lycopersicon esculentum Mill. cv. Nikita). Plants were subjected to moderate and severe levels of water stress (i.e. water potentials in the nutrient solution of-0.51 and -1.22 MPa, respectively).Water stress markedly affected the parameters of gas exchange. Net photosynthetic rate (Pn) decreased with the induction of water stress. Accordingly, a decrease in the transpiration rate (E) was observed. The ratio of both (Pn/E) resulted in a decrease in water use efficiency. One of the possible reasons for the reduction in Pn is structural damage to the thylakoids, which affects the photosynthetic transport of electrons. This was indicated by an increase in non-photochemical quenching and a reduction in the quantum yield of photosystem Ⅱ. Furthermore, a decrease in both leaf water potential and leaf osmotic potential was observed, which resulted in a significant osmotic adjustment during stress conditions. Analysis of the physiological responses was complemented with a study on changes in proline content. In stressed plants, a 10-fold increase in proline content was detected compared with control plants. It is clear that water stress tolerance is the result of a cumulative action of various physiological and biochemical processes, all of which were affected by PEG 6000-induced water stress.