Mounting evidence suggests that heat-shock proteins (HSPs) play a vital role in enhancing survival at high temperature. There is, however, considerable variation in patterns of HSP production among species, and even among and within individuals of a species. It is not known why this variation exists and to what extent variation in HSPs among organisms might be related to differences in thermotolerance. One possibility is that production of HSPs confers costs and natural selection has worked towards optimizing the cost-to-benefits of HSP synthesis and accumulation. However, the costs of this production have not been determined. If HSP production confers significant nitrogen (N) costs, then we reasoned that plants grown under low-N conditions might accumulate less HSP than high-N plants. Furthermore, if HSPs are related to thermotolerance, then variation in HSPs induced by N (or other factors) might correlate with variation in thermotolerance, here measured as short-term effects of heat stress on net CO assimilation and photosystem II (PSII) function. To test these predictions, we grew individuals of a single variety of corn (Zea mays L.) under different N levels and then exposed the plants to acute heat stress. We found that: (1) high-N plants produced greater amounts of mitochondrial Hsp60 and chloroplastic Hsp24 per unit protein than their low-N counterparts; and (2) patterns of HSP production were related to PSII efficiency, as measured by F /F . Thus, our results indicate that N availability influences HSP production in higher plants suggesting that HSP production might be resource-limited, and that among other benefits, chloroplast HSPs (e.g., Hsp24) may in some way limit damage to PSII function during heat stress.