A general model for carbon isotopes in red-lineage phytoplankton: Interplay between unidirectional processes and fractionation by RubisCO

The carbon isotopic composition of organic matter preserved in marine sediments provides a window into the global carbon cycle through geologic time, including variations in atmospheric CO_2 levels. Traditional models for interpreting isotope records of marine phytoplankton assume that these archives primarily reflect kinetic isotope discrimination by the carbon-fixing enzyme RubisCO. However, some in vivo and in vitro measurements appear to contradict this assumption, indicating that significant questions remain about the mechanistic underpinning of algal isotopic signatures, including the role of carbon concentrating mechanisms (CCMs). Here, we present a general model to explain photosynthetic carbon isotope fractionation (ε_P) in marine red-lineage phytoplankton groups; the model reproduces existing chemostat and batch culture datasets with a normalized root mean squared error (nRMSE) of 6.8%. Our framework proposes that a nutrient- and light-dependent step upstream of RubisCO is a kinetic barrier to carbon acquisition and therefore represents a significant source of isotopic fractionation. We suggest this step represents a carbon concentrating strategy that becomes favorable to cells under conditions of excess photon flux. The primary implications are that RubisCO is predicted to exert minimal isotopic control in photon-rich, nutrient-limited regimes but becomes influential as growth becomes light-limited. This framework enables both environment-specific and taxon-specific isotopic predictions. By refining the mechanistic understanding of marine photosynthetic carbon isotope fractionation, we may begin to reconcile existing datasets and reexamine Phanerozoic isotope records—including the resulting CO_2 reconstructions—by emphasizing the influence of different types of resource limitation on photosynthetic carbon acquisition

Carbon isotope ratioss of coccolith-associated polysaccharides of Emiliania huxleyi as a function of growth rate and CO2 concentration

The calcite plates, or coccoliths, of haptophyte algae including Emiliania huxleyi are formed in intracellular vesicles in association with water–soluble acidic polysaccharides. These coccolith–associated polysaccharides (CAPs) are involved in regulating coccolith formation and have been recovered from sediment samples dating back to ∼180 Ma. Paired measurements of the carbon isotopic compositions of CAPs and coccolith calcite have been proposed as a novel paleo–pCO2 barometer, but additional proxy validation and development are still required. Here we present culture results quantifying carbon isotopic offsets between CAPs and other cellular components: bulk organic biomass, alkenones, and calcite. E. huxleyi was grown in nitrate–limited chemostat experiments at growth rates (µ) of 0.20–0.62/d and carbon dioxide concentrations of 10.7–17.6 µmol/kg. We find that CAPs are isotopically enriched by 4.5–10.1‰ relative to bulk organic carbon, exhibiting smaller isotopic offsets at faster growth rates and lower CO2 concentrations. This variability suggests that CAPs record a complementary signature of past growth conditions with different sensitivity than alkenones or coccolith calcite. By measuring the isotopic compositions of all three molecules and minerals of self-consistent origin, the ratio µ/[CO2(aq)] may be reconstructed with fewer assumptions than current approaches

A general model for carbon isotopes in red-lineage phytoplankton: Interplay between unidirectional processes and fractionation by RubisCO

The carbon isotopic composition of organic matter preserved in marine sediments provides a window into the global carbon cycle through geologic time, including variations in atmospheric CO_2 levels. Traditional models for interpreting isotope records of marine phytoplankton assume that these archives primarily reflect kinetic isotope discrimination by the carbon-fixing enzyme RubisCO. However, some in vivo and in vitro measurements appear to contradict this assumption, indicating that significant questions remain about the mechanistic underpinning of algal isotopic signatures, including the role of carbon concentrating mechanisms (CCMs). Here, we present a general model to explain photosynthetic carbon isotope fractionation (ε_P) in marine red-lineage phytoplankton groups; the model reproduces existing chemostat and batch culture datasets with a normalized root mean squared error (nRMSE) of 6.8%. Our framework proposes that a nutrient- and light-dependent step upstream of RubisCO is a kinetic barrier to carbon acquisition and therefore represents a significant source of isotopic fractionation. We suggest this step represents a carbon concentrating strategy that becomes favorable to cells under conditions of excess photon flux. The primary implications are that RubisCO is predicted to exert minimal isotopic control in photon-rich, nutrient-limited regimes but becomes influential as growth becomes light-limited. This framework enables both environment-specific and taxon-specific isotopic predictions. By refining the mechanistic understanding of marine photosynthetic carbon isotope fractionation, we may begin to reconcile existing datasets and reexamine Phanerozoic isotope records—including the resulting CO_2 reconstructions—by emphasizing the influence of different types of resource limitation on photosynthetic carbon acquisition

CO_2-dependent carbon isotope fractionation in Archaea, Part I: Modeling the 3HP/4HB pathway

The 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) pathway of carbon fixation is found in thermophilic Crenarchaeota of the order Sulfolobales and in aerobic, ammonia-oxidizing Thaumarchaeota. Unlike all other known autotrophic carbon metabolisms, this pathway exclusively uses HCO_3- rather than CO_2 as the substrate for carbon fixation. Biomass produced by the 3HB/4HP pathway is relatively ^(13)C-enriched compared to biomass fixed by other autotrophic pathways, with total biosynthetic isotope effects (ε_(Ar)) of ca. 3‰ in the Sulfolobales and ca. 20‰ in the Thaumarchaeota. Explanations for the difference between these values usually invoke the dual effects of thermophily and growth at low pH (low [HCO_3-]) for the former group vs. mesophily and growth at pH > 7 (high [HCO_3-]) for the latter group. Here we examine the model taxa Metallosphaera sedula and Nitrosopumilus maritimususing an isotope flux-balance model to argue that the primary cause of different ε_(Ar) values more likely is the presence of carbonic anhydrase in M. sedula and its corresponding absence in N. maritimus. The results suggest that the pool of HCO_3-inside N. maritimus is out of isotopic equilibrium with CO_2 and that the organism imports < 10% HCO_3- from the extracellular environment. If correct and generalizable, the aerobic, ammonia-oxidizing marine Thaumarchaeota are dependent on passive CO_2 uptake and a slow rate of intracellular conversion to HCO_3-. Values of ε_(Ar) should therefore vary in response to growth rate (μ) and CO_2 availability, analogous to eukaryotic algae, but in the opposite direction: ε_(Ar) becomes smaller as [CO_(2(aq))] increases and/or μ decreases. Such an idea represents a testable hypothesis, both in the laboratory and in natural systems. Sensitivity to μ and CO_2 implies that measurements of ε_(Ar) may hold promise as a pCO_2 paleobarometer

Toward a Cenozoic history of atmospheric CO2

The geological record encodes the relationship between climate and atmospheric carbon dioxide (CO2) over long and short timescales, as well as potential drivers of evolutionary transitions. However, reconstructing CO2 beyond direct measurements requires the use of paleoproxies and herein lies the challenge, as proxies differ in their assumptions, degree of understanding, and even reconstructed values. In this study, we critically evaluated, categorized, and integrated available proxies to create a high-fidelity and transparently constructed atmospheric CO2 record spanning the past 66 million years. This newly constructed record provides clearer evidence for higher Earth system sensitivity in the past and for the role of CO2 thresholds in biological and cryosphere evolution