Metabolic Reliance on Photosynthesis Depends on Both Irradiance and Prey Availability in the Mixotrophic Ciliate, Strombidium cf. basimorphum

Many species of the ciliate genus Strombidium can acquire functional chloroplasts from a wide range of algal prey and are thus classified as generalist non-constitutive mixotrophs. Little, however, is known about the influence of irradiance and prey availability on their ability to exploit the photosynthetic potential of the chloroplasts, and how this may explain their spatial and temporal distribution in nature. In this study, inorganic carbon uptake, growth, and ingestion rates were measured for S. cf. basimorphum under three different irradiances (10, 40, and 120 μmol photons m–2 s–1) when acclimated to three different prey densities (5 × 103, 1 × 104, and 4 × 104 cells mL–1), as well as when allowed to deplete the prey. After prey depletion, cultures survived without prey longest (∼6 days) at the medium irradiance treatment (40 μmol photons m–2 s–1), while ciliate density, inorganic carbon uptake rates, and cellular chl-a content declined fastest at the highest irradiance treatment. This indicates that the ciliates may be unable to maintain the chloroplasts functionally without replacement at high irradiances. Ingestion rates were not shown to be significantly influenced by irradiance. The maximum gross growth efficiency (GGE) in this study (1.1) was measured in cultures exposed to the medium test irradiance and lowest prey density treatment (5 × 103 cells mL–1). The relative contribution of inorganic carbon uptake to the ciliate carbon budget was also highest in this treatment (42%). A secondary GGE peak (0.99) occurred when cultures were exposed to the highest test irradiance and the medium prey density. These and other results suggest that S. cf. basimorphum, and other generalist non-constitutive mixotrophs, can flexibly exploit many different environmental conditions across the globe.

Ecophysiological traits of mixotrophic Strombidium spp

Ciliates represent an important trophic link between nanoplankton and mesoplankton. Many species acquire functional chloroplasts from photosynthetic prey, being thus mixotrophs. Little is known about which algae they exploit, and of the relevance of inorganic carbon assimilation to their metabolism. To get insights into these aspects, laboratory cultures of three mixotrophic Strombidium spp. were established and 35 photosynthetic algal species were tested as prey. The relative contributions of ingestion and photosynthesis to total carbon uptake were determined, and responses to prey starvation were studied. Ciliate growth was supported by algal species in the 2–12 μm size range, with cryptophytes and chlorophytes being the best prey types. Inorganic carbon incorporation was only quantitatively important when prey concentration was low (3–100 μgCL−1), when it led to increased gross growth efficiencies. Chla specific inorganic carbon uptake rates were reduced by 60–90% compared to that of the photosynthetic prey. Inorganic carbon uptake alone could not sustain survival of cultures and ciliate populations declined by 25–30% during 5 days of starvation. The results suggest that mixotrophy in Strombidium spp. may substantially bolster the efficiency of trophic transfer when biomass of small primary producers is low.

External α-carbonic anhydrase and solute carrier 4 are required for bicarbonate uptake in a freshwater angiosperm

The freshwater monocot Ottelia alismoides is the only known species to operate three CO2-concentrating mechanisms (CCMs): constitutive bicarbonate (HCO3–) use, C4 photosynthesis, and facultative Crassulacean acid metabolism, but the mechanism of HCO3– use is unknown. We found that the inhibitor of an anion exchange protein, 4,4'-diisothio-cyanatostilbene-2,2'-disulfonate (DIDS), prevented HCO3– use but also had a small effect on CO2 uptake. An inhibitor of external carbonic anhydrase (CA), acetazolamide (AZ), reduced the affinity for CO2 uptake but also prevented HCO3– use via an effect on the anion exchange protein. Analysis of mRNA transcripts identified a homologue of solute carrier 4 (SLC4) responsible for HCO3– transport, likely to be the target of DIDS, and a periplasmic α-carbonic anhydrase 1 (α-CA1). A model to quantify the contribution of the three different pathways involved in inorganic carbon uptake showed that passive CO2 diffusion dominates inorganic carbon uptake at high CO2 concentrations. However, as CO2 concentrations fall, two other pathways become predominant: conversion of HCO3– to CO2 at the plasmalemma by α-CA1 and transport of HCO3– across the plasmalemma by SLC4. These mechanisms allow access to a much larger proportion of the inorganic carbon pool and continued photosynthesis during periods of strong carbon depletion in productive ecosystems.

Carbonic anhydrase is involved in benthic foraminiferal calcification

Marine calcification is an important component of the global carbon cycle. The mechanism by which some organisms take up inorganic carbon for the production of their shells or skeletons, however, remains only partly known. Although foraminifera are responsible for a large part of the global calcium carbonate production, the process by which they concentrate inorganic carbon is debated. Some evidence suggests that seawater is taken up and participates relatively unaltered in the process of calcification, whereas other results suggest the involvement of transmembrane transport and the activity of enzymes like carbonic anhydrase. Here, we tested whether inorganic carbon uptake relies on the activity of carbonic anhydrase using incubation experiments with the large benthic, symbiont-bearing foraminifer Amphistegina lessonii. Calcification rates, determined by the alkalinity anomaly method, showed that inhibition of carbonic anhydrase by acetazolamide (AZ) stopped most of the calcification process. Inhibition of photosynthesis by either 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (DCMU) or by incubating the foraminifera in the dark, also decreased calcification rates, but to a lesser degree than with AZ. Results from this study show that carbonic anhydrase plays a key role in biomineralization of Amphistegina lessonii and indicates that calcification of those large benthic foraminifera might, to a certain extent, benefit from ocean acidification.