Effects of reduced carbonic anhydrase activity on CO₂assimilation rates inSetaria viridis: a transgenic analysis

In C 4 species, the major -carbonic anhydrase (-CA) localized in the mesophyll cytosol catalyses the hydration of CO 2 to HCO 3 -, which phosphoenolpyruvate carboxylase uses in the first step of C 4 photosynthesis. To address the role of CA in C 4 photosynthesis, we generated transgenic Setaria viridis depleted in -CA. Independent lines were identified with as little as 13% of wild-type CA. No photosynthetic defect was observed in the transformed lines at ambient CO 2 partial pressure (pCO 2 ). At low pCO 2 , a strong correlation between CO 2 assimilation rates and CA hydration rates was observed. C 18 O 16 O isotope discrimination was used to estimate the mesophyll conductance to CO 2 diffusion from the intercellular air space to the mesophyll cytosol (g m ) in control plants, which allowed us to calculate CA activities in the mesophyll cytosol (C m ). This revealed a strong relationship between the initial slope of the response of the CO 2 assimilation rate to cytosolic pCO 2 (AC m ) and cytosolic CA activity. However, the relationship between the initial slope of the response of CO 2 assimilation to intercellular pCO 2 (AC i ) and cytosolic CA activity was curvilinear. This indicated that in S. viridis, mesophyll conductance may be a contributing limiting factor alongside CA activity to CO 2 assimilation rates at low pCO 2 .