Inorganic carbon acquisition in the acid-tolerant alga Chlorella kessleri.

The ability of the freshwater alga, Chlorella kessleri, to maintain a carbon concentrating mechanism when grown at acid pH was investigated. The alga grows over the pH range 4.0-9.0 and was found to take up bicarbonate and CO2 actively when grown at pH 6.0. However, when grown at acid pH (below 5.5), it does not have active CO2 uptake. The acidotolerant species maintained an internal pH of 6.1-7.5 over the external pH range 4.5-7.5, thus the pH difference between the cell interior and the external medium was large enough to allow for the diffusive uptake of CO2 at acid external pH. Mass spectrometric monitoring of O2 and CO2 fluxes by suspensions of C. kessleri, grown at acid pH, and maintained at pH 7.5 showed that the rates of O2 evolution did not exceed those of CO2 uptake. The final CO2 compensation concentrations of 14.0-17.7 µM reached by photosynthetic cells were above the CO2 equilibrium concentration in the external medium, indicating a lack of active CO2 uptake at acid pH. Chlorella kessleri accumulated CO2 with internal concentrations that were 9.9, 18.7 and 22.7-fold that of the external medium for cells grown, respectively, at pH 4.5, 5.0 and 5.5. The ability of C. kessleri cells to accumulate high intracellular concentrations of inorganic carbon at acid pH would provide a sufficiently high concentration of CO2 at the active site of Rubisco thus allowing the alga to maintain growth rates similar to those at alkaline pH.

Organization of astaxanthin within oil bodies of Haematococcus pluvialis studied with polarization-dependent harmonic generation microscopy.

Nonlinear optical microscopy was used to image the localization of astaxanthin accumulation in the green alga, Haematococcus pluvialis. Polarization-in, polarization-out (PIPO) second harmonic generation (SHG) and third harmonic generation (THG) microscopy was applied to study the crystalline organization of astaxanthin molecules in light-stressed H. pluvialis in vivo. Since astaxanthin readily forms H- and J-aggregates in aqueous solutions, PIPO THG studies of astaxanthin aggregates contained in red aplanospores were compared to PIPO THG of in vitro self-assembled H- and J-aggregates of astaxanthin. The PIPO THG data clearly showed an isotropic organization of astaxanthin in red aplanospores of H. pluvialis. This is in contrast to the highly anisotropic organization of astaxanthin in synthetic H- and J-aggregates, which showed to be uniaxial. Since carotenoids in vitro preferentially form H- and J-aggregates, but in vivo form a randomly organized structure, this implies that astaxanthin undergoes a different way of packing in biological organisms, which is either due to the unique physical environment of the alga or is controlled enzymatically.