CO
2 is used for carboxylation in chloroplast stroma, and diffuses in through stomata, intercellular air spaces, the cell wall, plasma membranes, and the chloroplast envelope. Stomatal resistance (
Rs) and mesophyll resistance (
Rm) occur in this pathway, the inverse of which are stomatal conductance (
gs) and mesophyll conductance (
gm), respectively, metrics used as indicators of conductivity.
gm is comparable to
gs and therefore important for photosynthesis. There are three ways to measure
gm: the chlorophyll fluorescence, curve-fitting, and stable carbon isotope methods. Here, we explain these methods and critical points in their respective measurements. The most reliable is the stable carbon isotope method, but parameter setting is difficult because small differences in parameters can result in large differences in
gm. We determined which parameters cause serious errors in estimating gm changes in response to CO
2. Errors in the CO
2 compensation point in the absence of mitochondrial respiration (
Γ*) and mitochondrial respiration in light (
Rd) exerted the most significant changes in gm at low CO
2 concentrations. Although these parameters are usually adopted from previous studies on limited numbers of species, they should be experimentally determined. Although calculating gm will still not be simple, precise measurements are required for understanding CO
2 diffusion in leaves.
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