Inorganic phosphorus (P-i) and carbon (here, CO2) potentially limit the photosynthesis of phytoplankton simultaneously (colimitation). A single P-i limitation generally reduces photosynthesis, but the effect of a colimitation is not known. Therefore, photosynthesis was measured under P-i-limited conditions and high and low CO2, and osmo-mixotrophic (i.e., growth in the presence of glucose) conditions that result in colimiting conditions in some cases. The green alga Chlamydomonas acidophila Negoro was used as a model organism because low P-i and CO2 concentrations likely influence its photosynthetic rates in its natural environment. Results showed a decreasing maximum photosynthetic rate (P-max) and maximum quantum yield (Theta(II)) with increasing P-i limitation. In addition, a P-i limitation enhanced the relative contribution of dark respiration to P-max (R-d:P-max) but did not influence the compensation light intensity. P-max positively correlated with the cellular RUBISCO content. Osmo-mixotrophic conditions resulted in similar P-max, Theta(II), and RUBISCO content as in high-CO2 cultures. The low-CO2 cultures were colimited by P-i and CO2 and had the highest P-max, Theta(II), and RUBISCO content. Colimiting conditions for P-i and CO2 in C. acidophila resulted in an enhanced mismatch between photosynthesis and growth rates compared to the effect of a single P- i limitation. Primary productivity of colimited phytoplankton could thus be misinterpreted.

Chlamydomonas acidophila, a dominant phytoplankton species in the very acidic Lake 111 (pH 2.7) situated in Germany, faces low concentrations of inorganic phosphorus (P-i), inorganic carbon (C-i) and potassium (K+) in its environment, which may lead to a complex colimitation by these nutrients. We performed laboratory and field investigations to test for P-i limitation and its dependence on C-i and K+ concentrations. The minimum cell quota for phosphorus (Q(0)) and phosphatase enzyme activity were similar to those for neutrophilic algae, despite the low pH and high concentrations of iron and aluminium, indicating no extra metabolic costs or inhibition of enzymes by the extreme environment. The threshold concentration of soluble reactive phosphorus for growth (SRPt), the algal C:P ratio and the alkaline phosphatase enzyme activity all suggested a moderate P-i limitation of C. acidophila in Lake 111. SRPt and Q(0) were higher at low CO2 and K+ concentrations in culture, showing a relationship between C-i and P-i acquisition. Furthermore, SRPt and Q(0) were higher under K+/P-i-colimiting conditions than under P-i-limiting conditions alone, suggesting that K+ concentrations influence P-i limitation in C. acidophila as well. Our results show that a limitation by one macronutrient requires consideration of the availability of the others as their uptake mechanisms depend on each other. Notwithstanding these interactions, C-i or K+ concentrations had no clear influence on the P-i limitation of C. acidophila in Lake 111.

Simultaneous limitation of plant growth by two or more nutrients is increasingly acknowledged as a common phenomenon in nature, but its cellular mechanisms are far from understood. We investigated the uptake kinetics of CO(2) and phosphorus of the algae Chlamydomonas acidophila in response to growth at limiting conditions of CO(2) and phosphorus. In addition, we fitted the data to four different Monod-type models: one assuming Liebigs Law of the minimum, one assuming that the affinity for the uptake of one nutrient is not influenced by the supply of the other (independent colimitation) and two where the uptake affinity for one nutrient depends on the supply of the other (dependent colimitation). In addition we asked whether the physiological response under colimitation differs from that under single nutrient limitation. We found no negative correlation between the affinities for uptake of the two nutrients, thereby rejecting a dependent colimitation. Kinetic data were supported by a better model fit assuming independent uptake of colimiting nutrients than when assuming Liebigs Law of the minimum or a dependent colimitation. Results show that cell nutrient homeostasis regulated nutrient acquisition which resulted in a trade-off in the maximum uptake rates of CO(2) and phosphorus, possibly driven by space limitation on the cell membrane for porters for the different nutrients. Hence, the response to colimitation deviated from that to a single nutrient limitation. In conclusion, responses to single nutrient limitation cannot be extrapolated to situations where multiple nutrients are limiting, which calls for colimitation experiments and models to properly predict growth responses to a changing natural environment. These deviations from single nutrient limitation response under colimiting conditions and independent colimitation may also hold for other nutrients in algae and in higher plants.

The extremophilic microalga Chlamydomonas acidophila inhabits very acidic waters (pH 2-3.5), where its growth is often limited by phosphorus (P) or colimited by P and inorganic carbon (CO(2)). Because this alga is a major food source for predators in acidic habitats, we studied its fatty acid content, which reflects their quality as food, grown under a combination of P-limited and different carbon conditions (either mixotrophically with light + glucose or at high or low CO(2), both without glucose). The fatty acid composition largely depended on the cellular P content: stringent P-limited cells had a higher total fatty acid concentration and had a lower percentage of polyunsaturated fatty acids. An additional limitation for CO(2) inhibited this decrease, especially reflected in enhanced concentrations of 18:3(9,12,15) and 16:4(3,7,10,13), resulting in cells relatively rich in polyunsaturated fatty acids under colimiting growth conditions. The percentage of polyunsaturated to total fatty acid content was positively related with maximum photosynthesis under all conditions applied. The two factors, P and CO(2), thus interact in their effect on the fatty acid composition in C. acidophila, and colimited cells P-limited algae can be considered a superior food source for herbivores because of the high total fatty acid content and relative richness in polyunsaturated fatty acids.