The accumulation of proline under environmental stress is a conserved response of plants. Five decades have passed since the first report of proline accumulation in plants (Barnnet and Naylor, 1966). Many hypotheses have been put forward regarding assignment of a function to proline. These proposed roles include antioxidant capacity, osmoprotection, signaling, developmental function, and contributing in redox and cellular homeostasis (Smirnoff and Cumbes, 1989; Mattioli et al., 2008; Szabados and Savoure, 2010; Kishor et al., 2015). Among these, the osmotic adjustment, osmoprotectant, and antioxidant role have probably been the earliest and most widely accepted functions of proline. It is known that compatible osmolytes contribute in the retention of water, but also some of them force proteins to adopt a compactly folded structure, preventing the unfolding and reducing the exposed surface of the protein to damaging compounds (Attri et al., 2010). In case of proline this was demonstrated both in vitro and in single cell organisms. For example, it was observed that proline overproducing E. coli mutants had greater osmotolerance (Csonka et al., 1988) and also proline can inhibit protein aggregation in vivo (Ignatova and Gierasch, 2006). However, neither osmotic adjustment nor osmoprotection has been clearly confirmed in plants (Maggio et al., 2002; Kavi Kishor and Sreenivasulu, 2014). This is probably more complex in plants because plants have different osmoregulatory mechanisms. Controversy is present regarding its proposed antioxidant role because recent evidence demonstrates that proline cannot scavenge singlet oxygen, superoxide, nitric oxide, peroxynitrite nor nitrogen dioxide (Signorelli et al., 2013, 2016). In view of the lack of activity in previously suggested roles it has recently been proposed that proline acts exclusively as a hydroxyl radical scavenger (Signorelli et al., 2014, 2015), but more evidence for the physiological importance of this role is needed.