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Clarisse Uwizeye

Quantitative imaging methods to investigate photosynthetic cells

Published on 16 December 2020
Thesis presented December 16, 2020

Abstract:
Phytoplankton is the group of photosynthetic microorganisms (microalgae and cyanobacteria) living in suspension in marine and fresh waters. Through photosynthesis, phytoplankton produce large amounts of the oxygen essential for marine and terrestrial life. Marine microalgae are also promising organisms for biotechnological applications (human and animal food, biofuels). Because of their ecological and economic importance, the study of the phytoplankton responses to environmental challenged (including the ones induced by human activity and global warming) is a developing field of research. Phytoplankton activity is influenced by changes in the vertical stratification of the water column, which modulate light energy availability as well as nutrient supply to phytoplankton cells in a temperature-dependent manner. Based on light and nutrient availability, phytoplankton cells have evolved different lifestyles: autotrophy (photosynthetic activity), mixotrophy (simultaneous use of photosynthesis and respiration of exogenous carbon sources for growth) and photosymbiosis (symbiotic interactions with animal cells).
In this thesis, I have studied the physiological responses of phytoplankton cells to environmental changes at the cellular and subcellular levels. To achieve this goal, I have developed a complete imaging workflow to perform quantitative morphometric analyses of entire algal cells, representatives of ecologically-successful and laboratory-model microalgal species. The protocol starts with FIB-SEM (Focused Ion Beam - Scanning Electron Microscopy) or SBF-SEM (Serial Block Facing – Scanning Electron Microscopy), to acquire high-resolution images. By implementing the 3D image analysis protocol, it is possible to obtained high-resolution whole cells models in three dimensions, suitable to perform quantitative analyses. Thanks to these tools, I have been able to image the adaptation of phytoplankton to various environmental conditions: i. changes in the size and morphology of plastids and mitochondria during light acclimation in diatoms, ii. changes in organelles interaction during nutrient acclimation in Nannochloropsis, iii. morphological changes occurring during photosymbiosis in Phaeocystis.
Overall, this work reveals several scenarios of phytoplankton acclimation at both the cellular and subcellular levels. I have also validated the use of this protocol in plants in a study on chloroplast biogenesis during de-etiolation in Arabidopsis of plastids.

Keywords:
3D imaging, FIB-SEM, Morphometric analyses, Phytoplankton, Photosymbiosis, Organelles