The exact relationship between polyamine, abscisic acid and proline metabolisms is still poorly understood. In the present study, the effects of putrescine and abscisic acid treatments alone or in combination with polyethylene glycol-induced osmotic stress were investigated in young wheat plants. It was observed that abscisic acid plays a role in the coordinated regulation of the proline and polyamine biosynthetic pathways, which compounds are related to each other through a common precursor. Abscisic acid pre-treatment induced similar alteration of polyamine contents as the osmotic stress, namely increased the putrescine, but decreased the spermidine contents in the leaves. These changes were mainly related to the polyamine cycle, as both the synthesis and peroxisomal oxidation of polyamines have been induced at gene expression level. Although abscisic acid and osmotic stress influenced the proline metabolism differently, the highest proline accumulation was observed in the case of abscisic acid treatments. The proline metabolism was partly regulated independently and not in an antagonistic manner from polyamine synthesis. Results suggest that the connection, which exists between polyamine metabolism and abscisic acid signalling leads to the controlled regulation and maintenance of polyamine and proline levels under osmotic stress conditions in wheat seedlings.