Glycine betaine (GB) is an effective compatible solute that improves the tolerance in plants to various stresses. We investigated the effects of 2 mM GB applied to the roots of a tobacco (Nicotiana tabacum L.) cultivar on enhancing photosynthesis under low-temperature (LT) stress (5/5 °C, 12/12 h, 300 µmol m-2 s-1) and in the subsequent recovery (25/18 °C) from the stress. The net photosynthetic rate, intrinsic efficiency measured as the ratio of variable to maximum fluorescence, and actual efficiency of the photochemistry of photosystem 2 as well as the ATPase activity in the thylakoid membrane decreased, and a distinct K step in the fluorescence transient O-J-I-P appeared under cold stress. Exogenous GB alleviated the decrease in all these parameters. The LT-stress induced the accumulation of 33-66 kDa polypeptides and decreased the proportion of unsaturated fatty acids in the thylakoid membrane. In plants subjected to LT-stress, GB protected these polypeptides from damage and enhanced the proportion of unsaturated fatty acids. An increase in non-radiative energy dissipation (NPQ) may be involved in the improvement of the function of the thylakoid membrane by GB since exogenous GB protected violaxanthin de-epoxidase and enhanced NPQ. and C. Wang ... [et al.].