Studies with the variable J method have reported that mesophyll conductance (gm ) rapidly decreases with increasing intercellular CO2 partial pressures (Ci ) or decreasing irradiance. Similar responses have been suggested with the online isotope discrimination method, although with less consistency. Here we show that even when the true gm is constant, the variable J method can produce an artefactual dependence of gm on Ci or irradiance similar to those reported in previous studies for any of the following factors: day respiration and chloroplastic CO2 photocompensation point are estimated with Laisk method; Ci or electron transport rate is positively biased; net photosynthetic rate is negatively biased; insufficient NADPH is assumed while insufficient ATP limits RuBP regeneration. The isotopic method produces similar artefacts if fractionation of carboxylation or Ci is positively biased or Δ(13) negatively biased. A non-zero chloroplastic resistance to CO2 movement results in a qualitatively different dependence of gm on Ci or irradiance and this dependence is only sensitive at low Ci . We thus cannot rule out the possibility that previously reported dependence of gm on Ci or irradiance is a methodological artefact. Recommendations are made to take advantage of sensitivities of the variable J and isotopic methods for estimating gm .
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