Rubber production in guayule (Parthenium argentatum, Gray) is highest in the winter months. It has been suggested that lower nighttime temperatures stimulate rubber production. Higher rubber production could be the result of up regulation of the enzymes involved in rubber synthesis, or acclimation of the photosynthetic apparatus to cold temperatures, making photosynthates available for rubber production when the plants are not actively growing, or a combination of the two factors. The effect of low night temperatures on photosynthesis and rubber production was examined in greenhouse grown plants. The experiment was set up as a split plot with different sub plots enclosed in heated or non-heated containers during the night. Those placed in non-heated containers were exposed to the ambient night temperatures of the greenhouse. During the growing season, growth was measured as change in height, and midday and diurnal photosynthetic measurements were collected. At harvest fresh and dry weight of both roots and shoots were taken. Resin and rubber were then extracted with acetone and cyclohexane respectively. Midday photosynthesis was as high or higher in cold night plants than in the warm night plants. In addition, the cold night treatment had lower growth than the warm night treatment. Percent resin and rubber were significantly higher in both shoots and roots of cold night plants, as was the overall yield of rubber. An increase in photosynthetic rate in the cold night treatment, without the concomitant increase in growth seen in the warm night treatment, could account for the increase in rubber production during the winter months.