To investigate the role of glycine betaine in photosynthesis under stress, a transgenic wheat (Triticum aestivum L.) line T6 overaccumulating glycine betaine and its wild type Shi4185 were used. Seedlings were exposed to conditions of drought (30%, PEG-6000), heat (40°C) and their combination. The results revealed ultrastructural damage to the chloroplast and thylakoid lamellae with the withered phenotype by both drought and heat stress, and the damage was exacerbated by the combination of drought and heat. The appearance of a K step in the typical O-J-I-P curve and the decrease of Hill activity indicated a reduction of oxygen evolving complex function caused by stress. The greater damage was found in wild type than T6. Overaccumulation of glycine betaine in T6 could protect lipids in the thylakoid membrane from damage and stabilize the index of unsaturated fatty acids under stress. A lower ratio of monogalactosyl diacylglycerol/digalactosyl diacylglycerol and higher phosphatidylglycerol content in the thylakoid membrane of T6 were also observed under stress. These effects can promote stability of the thylakoid membrane. Otherwise, glycine betaine overaccumulation decreased photoinhibition of PSII under stress. The results also suggest that xanthophyll cycle-dependent non-radiative energy dissipation may be involved in the GB-mediated effects on PSII function under stress conditions. and G. P. Wang ... [et al.].