The transition of the physical phase of lipids in membrane fragments of a blue-green alga Anacystis nidulans was studied by a spin labeling technique. The maximum hyperfine splitting of the electron spin resonance spectrum of the N-oxyl -4', 4'-dimethyloxazolidine derivative of 5 -ketostearic acid plotted against the reciprocal of the absolute temperature gave a discontinuity point that was characteristic of a transition of the physical phase of the hydrocarbon region of membrane lipids. The phase transition appeared at approximately 13 or 24 C in the organisms grown at 28 or 38 C, respectively.The and state 2 shift is dependent on the physical state of membrane lipids. In the chloroplasts of lettuce and spinach, on the other hand, there was no break in the Arrhenius plot of the electron transport reactions or of Mg'--induced changes of chlorophyll a fluorescence.It is suggested that the transitions of the hyperfine splitting of the ESR signal, electron transport, and the configurational change, as well as the appearance of the maximum of chlorophyll a fluorescence, in the thylakoid membranes of Anacystis nidulans are all related to the transition of the physical phase of membrane lipids between the liquid crystalline state and the mixed liquid crystal-solid state. It has been well demonstrated that the physical phase of lipids of biological membranes plays an important role in the physiological function of membranes (45,47). The proper functioning of biological membranes requires the presence of the smectic liquid crystalline state in which rotational and translational movements of lipid and protein molecules in the membrane are possible (29). It has been established in model membranes that the temperature of phase transition between the liquid crystalline and the solid states depends on the lipid species as well as the fatty acid composition (7,10,27,57); the higher the degree of unsaturation of fatty acids, the lower the phase transition temperature. The same relationship between the phase transition temperature and fatty acid composition also appears in the biological membranes of Escherichia coli (44,50,59), Mycoplasma laidlawii (14, 35, 55, 56), mitochondria (30, 32, 47), and yeast (13). Drastic changes of physiological activities of the biological membranes are observed at the phase transition temperatures. This is seen as changes in growth, transport, and respiration in E. coli (44,59) and in phosphorylation, respiration, and conformational changes in mitochondria (24,31,32,48