Abstract
Microalgae are the richest source of natural carotenoids—accessory photosynthetic pigments used as natural antioxidants, safe colorants, and nutraceuticals. Microalga Bracteacoccus aggregatus IPPAS C-2045 responds to stresses, including high light, with carotenogenesis—gross accumulation of secondary carotenoids (the carotenoids structurally and energetically uncoupled from photosynthesis). Precise mechanisms of cytoplasmic transport and subcellular distribution of the secondary carotenoids under stress are still unknown. Using multimodal imaging combining micro-Raman imaging (MRI), fluorescent lifetime (τ) imaging (FLIM), and transmission electron microscopy (TEM), we monitored ultrastructural and biochemical rearrangements of B. aggregatus cells during the stress-induced carotenogenesis. MRI revealed a decline in the diversity of molecular surrounding of the carotenoids in the cells compatible with the relocation of the bulk of the carotenoids in the cell from functionally and structurally heterogeneous photosynthetic apparatus to the more homogenous lipid matrix of the oleosomes. Two-photon FLIM highlighted the pigment transformation in the cell during the stress-induced carotenogenesis. The structures co-localized with the carotenoids with shorter τ (mainly chloroplast) shrunk, whereas the structures harboring secondary carotenoids with longer τ (mainly oleosomes) expanded. These changes were in line with the ultrastructural data (TEM). Fluorescence of B. aggregatus carotenoids, either in situ or in acetone extracts, possessed a surprisingly long lifetime. We hypothesize that the extension of τ of the carotenoids is due to their aggregation and/or association with lipids and proteins. The propagation of the carotenoids with prolonged τ is considered to be a manifestation of the secondary carotenogenesis suitable for its non-invasive monitoring with multimodal imaging.
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The datasets generated during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
The TEM studies were carried out at the Shared Research Facility “Electron microscopy in life sciences” at Moscow State University (Unique Equipment “Three-dimensional electron microscopy and spectroscopy”). Photosynthetic pigment assay was done using the Shared Research Facility “Phototrophic Organism Phenotyping.” The authors are indebted to Dr. Dmitry Kochkin for his assistance with carotenoid profiling. The authors are indebted to Dr. Olga Baulina for her assistance with TEM sample preparation.
Funding
This work was partially supported by the Russian Science Foundation (grants 23–44-00006, cultivation of microalgae; 23–74-00037, biochemical analyses). N.N.S. acknowledges the support of the Ministry of Science and Higher Education of the Russian Federation.
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Solovchenko, A., Lobakova, E., Semenov, A. et al. Multimodal non-invasive probing of stress-induced carotenogenesis in the cells of microalga Bracteacoccus aggregatus. Protoplasma (2024). https://doi.org/10.1007/s00709-024-01956-9
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DOI: https://doi.org/10.1007/s00709-024-01956-9