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Summary In this project, we hypothesize that cotton ( Gossypium hirsutum ) leaf temperature and the responses of leaf photosynthesis to temperature will change as the leaves expand and that differences between young and mature leaves will be associated with the proportion of saturated fatty acids in thylakoid and other membrane lipids. To that end, we studied main stem leaves obtained from plants growing in a temperature controlled greenhouse and at different times in the field season. We found that young leaves (∼5 d old) had higher mid day temperatures, lower stomatal conductance and higher thermal optima as measured by Φ PSII temperature curves than did more mature leaves (∼13 d old). Young leaves also had significant differences in fatty acid saturation with the warmer, young leaves having a higher proportion of palmitic acid (16:0) and lower linoleic acid (18:3) in total lipid extracts and higher 16:0 and lower palmitoleic acid (16:1) in the chloroplast membrane phosphoglycerides, digalactosyldiacylglycerol (in the greenhouse) and phosphatidylglycerol when compared with cooler, more mature leaves. Later in the growing season, leaf temperature, stomatal conductance and Φ PSII temperature curves for young and more mature leaves were similar and the proportion of 16:0 fatty acids decreased and 16:1 increased in phosphatidylglycerol. We conclude that changes in temperature as cotton leaves expand leads to alterations in the fatty acid composition of thylakoid and other membranes and, consequently, influence photosynthesis/temperature responses.