PT  - JOURNAL ARTICLE
AU  - Nicky Atkinson
AU  - Yuwei Mao
AU  - Kher Xing Chan
AU  - Alistair J. McCormick
TI  - Condensation of Rubisco into a proto-pyrenoid in higher plant chloroplasts
AID  - 10.1101/2020.10.26.354332
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.10.26.354332
4099  - http://biorxiv.org/content/early/2020/10/27/2020.10.26.354332.short
4100  - http://biorxiv.org/content/early/2020/10/27/2020.10.26.354332.full
AB  - Photosynthetic CO2 fixation in plants is limited by the inefficiency of the CO2-assimilating enzyme Rubisco (D-ribulose-1,5-bisphosphate carboxylase/ oxygenase)1–3. In plants possessing the C3 pathway, which includes most major staple crops, Rubisco is typically evenly distributed throughout the chloroplast stroma. However, in almost all eukaryotic algae Rubisco aggregates within a microcompartment known as the pyrenoid, in association with a CO2-concentrating mechanism that improves photosynthetic operating efficiency under conditions of low inorganic carbon4. Recent work has shown that the pyrenoid matrix is a phase-separated, liquid-like condensate5. In the alga Chlamydomonas reinhardtii, condensation is mediated by two components: Rubisco and the linker protein EPYC1 (Essential Pyrenoid Component 1)6,7. Here we show that expression of mature EPYC1 and a plant-algal hybrid Rubisco leads to spontaneous condensation of Rubisco into a single phase-separated compartment in Arabidopsis chloroplasts, with liquid-like properties similar to a pyrenoid matrix. The condensate displaces the thylakoid membranes and is enriched in hybrid Rubisco containing the algal Rubisco small subunit required for phase separation. Promisingly, photosynthetic CO2 fixation and growth is not impaired in stable transformants compared to azygous segregants. These observations represent a significant initial step towards enhancing photosynthesis in higher plants by introducing an algal CO2-concentrating mechanism, which is predicted to significantly increase the efficiency of photosynthetic CO2 uptake8,9.Competing Interest StatementThe authors have declared no competing interest.