skip to main content

This content will become publicly available on May 19, 2023

Title: Modelling the pyrenoid-based CO2-concentrating mechanism provides insights into its operating principles and a roadmap for its engineering into crops
Many eukaryotic photosynthetic organisms enhance their carbon uptake by supplying concentrated CO2 to the CO2-fixing enzyme Rubisco in an organelle called the pyrenoid. Ongoing efforts seek to engineer this pyrenoid-based CO2-concentrating mechanism (PCCM) into crops to increase yields. Here we develop a computational model for a PCCM on the basis of the postulated mechanism in the green alga Chlamydomonas reinhardtii. Our model recapitulates all Chlamydomonas PCCM-deficient mutant phenotypes and yields general biophysical principles underlying the PCCM. We show that an effective and energetically efficient PCCM requires a physical barrier to reduce pyrenoid CO2 leakage, as well as proper enzyme localization to reduce futile cycling between CO2 and HCO3−. Importantly, our model demonstrates the feasibility of a purely passive CO2 uptake strategy at air-level CO2, while active HCO3− uptake proves advantageous at lower CO2 levels. We propose a four-step engineering path to increase the rate of CO2 fixation in the plant chloroplast up to threefold at a theoretical cost of only 1.3 ATP per CO2 fixed, thereby offering a framework to guide the engineering of a PCCM into land plants.
Authors:
Award ID(s):
1935444
Publication Date:
NSF-PAR ID:
10334841
Journal Name:
Nature plants
Volume:
8
Issue:
5
Page Range or eLocation-ID:
583–595
ISSN:
2055-0278
Sponsoring Org:
National Science Foundation
More Like this
  1. Approximately one-third of the Earth’s photosynthetic CO 2 assimilation occurs in a pyrenoid, an organelle containing the CO 2 -fixing enzyme Rubisco. How constituent proteins are recruited to the pyrenoid and how the organelle’s subcompartments—membrane tubules, a surrounding phase-separated Rubisco matrix, and a peripheral starch sheath—are held together is unknown. Using the model alga Chlamydomonas reinhardtii , we found that pyrenoid proteins share a sequence motif. We show that the motif is necessary and sufficient to target proteins to the pyrenoid and that the motif binds to Rubisco, suggesting a mechanism for targeting. The presence of the Rubisco-binding motif on proteins that localize to the tubules and on proteins that localize to the matrix–starch sheath interface suggests that the motif holds the pyrenoid’s three subcompartments together. Our findings advance our understanding of pyrenoid biogenesis and illustrate how a single protein motif can underlie the architecture of a complex multilayered phase-separated organelle.
  2. Abstract Different intensities of high temperatures affect the growth of photosynthetic cells in nature. To elucidate the underlying mechanisms, we cultivated the unicellular green alga Chlamydomonas reinhardtii under highly controlled photobioreactor conditions and revealed systems-wide shared and unique responses to 24-hour moderate (35°C) and acute (40°C) high temperatures and subsequent recovery at 25°C. We identified previously overlooked unique elements in response to moderate high temperature. Heat at 35°C transiently arrested the cell cycle followed by partial synchronization, up-regulated transcripts/proteins involved in gluconeogenesis/glyoxylate-cycle for carbon uptake and promoted growth. But 40°C disrupted cell division and growth. Both high temperatures induced photoprotection, while 40°C distorted thylakoid/pyrenoid ultrastructure, affected the carbon concentrating mechanism, and decreased photosynthetic efficiency. We demonstrated increased transcript/protein correlation during both heat treatments and hypothesize reduced post-transcriptional regulation during heat may help efficiently coordinate thermotolerance mechanisms. During recovery after both heat treatments, especially 40°C, transcripts/proteins related to DNA synthesis increased while those involved in photosynthetic light reactions decreased. We propose down-regulating photosynthetic light reactions during DNA replication benefits cell cycle resumption by reducing ROS production. Our results provide potential targets to increase thermotolerance in algae and crops.
  3. Elger, Kirsten ; Carlson, David ; Klump, Jens ; Peng, Ge (Ed.)
    Air-sea flux of carbon dioxide (CO2) is a critical component of the global carbon cycle and the climate system with the ocean removing about a quarter of the CO2 emitted into the atmosphere by human activities over the last decade. A common approach to estimate this net flux of CO2 across the air-sea interface is the use of surface ocean CO2 observations and the computation of the flux through a bulk parameterization approach. Yet, the details for how this is done in order to arrive at a global ocean CO2 uptake estimate varies greatly, unnecessarily enhancing the uncertainties. Here we reduce some of these uncertainties by harmonizing an ensemble of products that interpolate surface ocean CO2 bservations to near global coverage. We propose a common methodology to fill in missing areas in the products and to calculate fluxes and present a new estimate of the net flux. The ensemble data product, SeaFlux (Gregor & Fay (2021), doi.org/10.5281/zenodo.4133802, https://github.com/luke-gregor/SeaFlux), accounts for the diversity of the underlying mapping methodologies. Utilizing six 30 global observation-based mapping products (CMEMS-FFNN, CSIR-ML6, JENA-MLS, JMA-MLR, MPI-SOMFFN, NIESFNN), the SeaFlux ensemble approach adjusts for methodological inconsistencies in flux calculations that can result in an average error of 15%more »in global mean flux estimates. We address differences in spatial coverage of the surface ocean CO2 between the mapping products which ultimately yields an increase in CO2 uptake of up to 19% for some products. Fluxes are calculated using three wind products (CCMPv2, ERA5, and JRA55). Application of an appropriately scaled gas exchange 35 coefficient has a greater impact on the resulting flux than solely the choice of wind product. With these adjustments, we derive an improved ensemble of surface ocean pCO2 and air-sea carbon flux estimates. The SeaFlux ensemble suggests a global mean uptake of CO2 from the atmosphere of 1.92 +/- 0.35 PgC yr-1. This work aims to support the community effort to perform model-data intercomparisons which will help to identify missing fluxes as we strive to close the global carbon budget.« less
  4. Although cyanobacteria and algae represent a small fraction of the biomass of all primary producers, their photosynthetic activity accounts for roughly half of the daily CO2 fixation that occurs on Earth. These microorganisms are able to accomplish this feat by enhancing the activity of the CO2-fixing enzyme Rubisco using biophysical CO2 concentrating mechanisms (CCMs). Biophysical CCMs operate by concentrating bicarbonate and converting it into CO2 in a compartment that houses Rubisco (in contrast with other CCMs that concentrate CO2 via an organic intermediate, such as malate in the case of C4 CCMs). This activity provides Rubisco with a high concentration of its substrate, thereby increasing its reaction rate. The genetic engineering of a biophysical CCM into land plants is being pursued as a strategy to increase crop yields. This review focuses on the progress toward understanding the molecular components of cyanobacterial and algal CCMs, as well as recent advances toward engineering these components into land plants.
  5. ABSTRACT Fish in coastal ecosystems can be exposed to acute variations in CO2 of between 0.2 and 1 kPa CO2 (2000–10,000 µatm). Coping with this environmental challenge will depend on the ability to rapidly compensate for the internal acid–base disturbance caused by sudden exposure to high environmental CO2 (blood and tissue acidosis); however, studies about the speed of acid–base regulatory responses in marine fish are scarce. We observed that upon sudden exposure to ∼1 kPa CO2, European sea bass (Dicentrarchus labrax) completely regulate erythrocyte intracellular pH within ∼40 min, thus restoring haemoglobin–O2 affinity to pre-exposure levels. Moreover, blood pH returned to normal levels within ∼2 h, which is one of the fastest acid–base recoveries documented in any fish. This was achieved via a large upregulation of net acid excretion and accumulation of HCO3− in blood, which increased from ∼4 to ∼22 mmol l−1. While the abundance and intracellular localisation of gill Na+/K+-ATPase (NKA) and Na+/H+ exchanger 3 (NHE3) remained unchanged, the apical surface area of acid-excreting gill ionocytes doubled. This constitutes a novel mechanism for rapidly increasing acid excretion during sudden blood acidosis. Rapid acid–base regulation was completely prevented when the same high CO2 exposure occurred in seawater with experimentally reduced HCO3− and pH, probably because reducedmore »environmental pH inhibited gill H+ excretion via NHE3. The rapid and robust acid–base regulatory responses identified will enable European sea bass to maintain physiological performance during large and sudden CO2 fluctuations that naturally occur in coastal environments.« less