Pyrenoid loss impairs carbon-concentrating mechanism induction and alters primary metabolism in Chlamydomonas reinhardtii

Carbon-concentrating mechanisms (CCMs) enable efficient photosynthesis and growth in CO2-limiting environments, and in eukaryotic microalgae localisation of Rubisco to a microcompartment called the pyrenoid is key. In the model green alga Chlamydomonas reinhardtii, Rubisco preferentially relocalises to the pyrenoid during CCM induction and pyrenoid-less mutants lack a functioning CCM and grow very poorly at low CO2. The aim of this study was to investigate the CO2 response of pyrenoid-positive (pyr+) and pyrenoid-negative (pyr–) mutant strains to determine the effect of pyrenoid absence on CCM induction and gene expression. Shotgun proteomic analysis of low-CO2-adapted strains showed reduced accumulation of some CCM-related proteins, suggesting that pyr– has limited capacity to respond to low-CO2 conditions. Comparisons between gene transcription and protein expression revealed potential regulatory interactions, since Rubisco protein linker (EPYC1) protein did not accumulate in pyr– despite increased transcription, while elements of the LCIB/LCIC complex were also differentially expressed. Furthermore, pyr− showed altered abundance of a number of proteins involved in primary metabolism, perhaps due to the failure to adapt to low CO2. This work highlights two-way regulation between CCM induction and pyrenoid formation, and provides novel candidates for future studies of pyrenoid assembly and CCM function

RF magnetron sputtering of Bi₁₂TiO₂₀ thin films on various substrates

Abstract Using RF magnetron sputtering, we deposited Bi12TiO20 (BTO) thin films on various substrates (glass, quartz, stainless steel (SS304), (001) and (111) Si and sital-ceramics, Al and Cu foils). The films had a constant thickness of 1.3 μm. The as-deposited films were studied by Raman spectroscopy, scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX), as well as by grazing incidence X-ray diffractometry (GIXRD). The GIXRD results reveal that the films are amorphous, while the Bi/Ti ratio varies between 9.5/1 and 11.8/1, as shown by the energy dispersive X-ray analysis (EDX). Further, the films deposited on glass and SS304 substrates were modified by laser irradiation (CuBr laser with a wavelength λ = 511 nm) to obtain an ordered cubic phase. The films deposited on quartz and on (001) and (111) Si substrates were thermally annealed at about 510 oC for three hours in ambient atmosphere. The modified films were characterized by Raman spectroscopy, scanning electron microscopy (SEM), EDX and X-ray diffractometry.</jats:p

Rubisco small-subunit α-helices control pyrenoid formation in Chlamydomonas.

International audienceThe pyrenoid is a subcellular microcompartment in which algae sequester the primary carboxylase, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The pyrenoid is associated with a CO(2)-concentrating mechanism (CCM), which improves the operating efficiency of carbon assimilation and overcomes diffusive limitations in aquatic photosynthesis. Using the model alga Chlamydomonas reinhardtii, we show that pyrenoid formation, Rubisco aggregation, and CCM activity relate to discrete regions of the Rubisco small subunit (SSU). Specifically, pyrenoid occurrence was shown to be conditioned by the amino acid composition of two surface-exposed α-helices of the SSU: higher plant-like helices knock out the pyrenoid, whereas native algal helices establish a pyrenoid. We have also established that pyrenoid integrity was essential for the operation of an active CCM. With the algal CCM being functionally analogous to the terrestrial C(4) pathway in higher plants, such insights may offer a route toward transforming algal and higher plant productivity for the future