Photosystem Ⅱ membrane was reconstituted with phosphatidylcholine (PC) with different kinds of fatty acyl chains and the protection of PC to photosystem Ⅱ (PSⅡ) membrane during heat treatment was investigated using oxygen electrode, variable fluorescence and circular dichroism (CD) spectroscopy. Heat treatment decreased the oxygen evolution rate and the F′v/Fm′ ratio of PSⅡ membrane and influenced CD spectra of PSⅡ membrane, but PC inhibited the effect of heat treatment on the oxygen evolution rate, the F′v/F′m ratio and CD spectra of PSⅡ membrane. The results indicate that PC can protect PSⅡ membrane against heat treatment and the alterations in the unsaturated fatty acid extent in PC can cause the changes of the protection ability.