Concurrent Measurement of O₂ Production and Isoprene Emission During Photosynthesis: Pros, Cons and Metabolic Implications of Responses to Light, CO₂ and Temperature

ABSTRACT Traditional leaf gas exchange experiments have focused on net CO 2 exchange ( A net ). Here, using California poplar ( Populus trichocarpa ), we coupled measurements of net oxygen production (NOP), isoprene emissions and δ 18 O in O 2 to traditional CO 2 /H 2 O gas exchange with chlorophyll fluorescence, and measured light, CO 2 and temperature response curves. This allowed us to obtain a comprehensive picture of the photosynthetic redox budget including electron transport rate (ETR) and estimates of the mean assimilatory quotient (AQ = A net /NOP). We found that A net and NOP were linearly correlated across environmental gradients with similar observed AQ values during light (1.25 ± 0.05) and CO 2 responses (1.23 ± 0.07). In contrast, AQ was suppressed during leaf temperature responses in the light (0.87 ± 0.28), potentially due to the acceleration of alternative ETR sinks like lipid synthesis. A net and NOP had an optimum temperature ( T opt ) of 31°C, while ETR and δ 18 O in O 2 (35°C) and isoprene emissions (39°C) had distinctly higher T opt . The results confirm a tight connection between water oxidation and ETR and support a view of light‐dependent lipid synthesis primarily driven by photosynthetic ATP/NADPH not consumed by the Calvin–Benson cycle, as an important thermotolerance mechanism linked with high rates of (photo)respiration and CO 2 /O 2 recycling.