Cold stress is among the environmental stressors limiting productivity, yield and quality of agricultural plants. Tolerance to cold stress is associated with the increased unsaturated fatty acids ratio in the plant membranes which are also known to be substrates of octadecanoid pathway for jasmonate and other oxylipins biosynthesis. Accumulation of osmoprotectant, glycine betaine (GB) is well known to be effective in the protecting membranes and mitigating cold stress effects but, the mode of action is poorly understood. We studied the role of GB in cold stress responses of two tomato cultivated varieties; Gerry (cold stress sensitive) and T47657 (moderately cold stress tolerant) and compared the differences in lypoxygenase-13 (TomLOXF) and fatty acid desaturase 7 (FAD7) gene expression profiles and physiological parameters including relative growth rates, relative water content, osmotic potential, photosynthetic efficiency, membrane leakage, lipid peroxidation levels. Our results indicated that GB might have a role in inducing FAD7 and LOX expressions for providing protection against cold stress in tomato plants which could be related to the desaturation process of lipids leading to increased membrane stability and/or induction of other genes related to stress defense mechanisms via octadecanoid pathway or lipid peroxidation products.