Mass Spectrometry Analysis of Oxygen Gas Exchange in High- and LOW-CO2 Cells of Chlorella vulgaris

Abstract

Abstract Oxygen gas exchange was monitored in the unicellular green alga Chlorella vulgaris 211 - 11 h by means of a mass spectrometer equipped with a special membrane gas-inlet-system and a photosynthetic reaction vessel. CO2-dependent 18O2-uptake as well as 16O2-evolution were analyzed in both High- and Low-CO2 cells. In High-CO2 cells, the 18O2-ruptake in the light (UL) decreased by 65% upon addition of 3 mᴍ NaHCO3 , while 16O2-evolution (E) was increased approx. 1.8 times by the same treatment. 18O2-uptake in the dark (UD) was not affected by the addition of external inorganic carbon (Ci). The addition of 3.3 mᴍ NaHCO3 also affected UL and E in Low CO2-cells, however, to a minor extent. UL under CO2-saturating conditions was light intensity-independent up to 2 klux and 1.2 klux in High- and Low-CO2 cells, respectively. Above these light intensities UL increased approx. 4-fold in High- and approx. 6-fold in Low-CO2 cells. Under CO2-limiting conditions, however, UL increased in High-CO2 cells even under very low light intensities, showing that photorespiratory oxygen uptake occurred even in the near vicinity of the light compensation point. Under C02-saturating and strong light conditions UL represented almost half of E in Low-CO2 cells and about 30 % of E in High-CO2 cells. In Low-CO2 cells addition of ethoxyzolamide (EZA), an inhibitor of carbonic anhydrase, enhanced UL and suppressed E and NET under CO2-limiting conditions, whereas the compound had only a minor effect on High-CO2 cells. DCMU (3 μᴍ) strongly inhibited E and UL under CO2-saturating conditions, with the remaining UL being smaller than UD . KCN (1 mᴍ) and SHAM (1.5 mᴍ) added to DCMU-treated Low-CO2 cells suppressed UL by approx. 50 % . The resulting value corresponded to half of UD . KCN also inhibited E under CO2-saturating conditions, with UL being strongly enhanced showing a maximal uptake at 0.4 mᴍ KCN . Under these conditions NET was nearly zero. The effect seems to be due to an inhibition of RubisCO and an enhancement of Mehler reactions. At 0.7 mᴍ KCN , DCMU entirely inhibited UL , but oxygen uptake appeared increased after turning the light off. This uptake corresponded to approx. 60 % of UD . Whereas KCN and SHAM inhibited approx. 70 % of UD , only 16% of UL was suppressed. These results suggest that the contribution of mitochondrial respiration to UL was negligeable, since UL seemed to be suppressed in the light under CO2-saturated conditions. Iodoacetamide, which is an inhibitor of the Calvin cycle and thereby diverts carbon into the respiratory pathway, inhibited E and NET under CO2-saturating conditions, but did not affect UL . This result also shows that UL is not due to mitochondrial respiration. A hydroxylamine derivative [20, 21] which changes the ratio of the RuBP carboxylation to oxygenation activity in tobacco leaves did not affect this ratio in Chlorella.