In vivo phosphoenolpyruvate carboxylase activity is controlled by CO 2 and O 2 mole fractions and represents a major flux at high photorespiration rates

Abstract

Phosphenolpyruvate carboxylase (PEPC)‐catalysed fixation of bicarbonate to C4 acids is commonly believed to represent a rather small flux in illuminated leaves. In addition, its potential variation with O2 and CO2 is not documented and thus is usually neglected in gas‐exchange studies. Here, we used quantitative NMR analysis of sunflower leaves labelled with 13CO2 (99% 13C) under controlled conditions and measured the amount of 13C found in the four C‐atom positions in malate, the major product of PEPC activity. We found that amongst malate 13C‐isotopomers present after labelling, most molecules were labelled at both C‐1 and C‐4, showing the incorporation of 13C at C‐4 by PEPC fixation and subsequent redistribution to C‐1 by fumarase (malate–fumarate equilibrium). In addition, absolute quantification of 13C content showed that PEPC fixation increased at low CO2 or high O2, and represented up to 1.8 μmol m−2 s−1, that is, 40% of net assimilation measured by gas exchange under high O2/CO2 conditions. Our results show that PEPC fixation represents a quantitatively important CO2‐fixing activity that varies with O2 and/or CO2 mole fraction and this challenges the common interpretation of net assimilation in C3 plants, where PEPC activity is often disregarded or considered to be constant at a very low rate.