Plastid thylakoid architecture optimizes photosynthesis in diatoms

Photosynthesis is a unique process that allows independent colonization of the land by plants and of the oceans by phytoplankton. Although the photosynthesis process is well understood in plants, we are still unlocking the mechanisms evolved by phytoplankton to achieve extremely efficient photosynthesis. Here, we combine biochemical, structural and in vivo physiological studies to unravel the structure of the plastid in diatoms, prominent marine eukaryotes. Biochemical and immunolocalization analyses reveal segregation of photosynthetic complexes in the loosely stacked thylakoid membranes typical of diatoms. Separation of photosystems within subdomains minimizes their physical contacts, as required for improved light utilization. Chloroplast 3D reconstruction and in vivo spectroscopy show that these subdomains are interconnected, ensuring fast equilibration of electron carriers for efficient optimum photosynthesis. Thus, diatoms and plants have converged towards a similar functional distribution of the photosystems although via different thylakoid architectures, which likely evolved independently in the land and the ocean.ISSN:2041-172

A systems-wide understanding of photosynthetic acclimation in algae and higher plants

The ability of phototrophs to colonise different environments relied on the robust protection against oxidative stress in phototrophs, a critical requirement for the successful evolutionary transition from water to land. Photosynthetic organisms have developed numerous strategies to adapt their photosynthetic apparatus to changing light conditions in order to optimise their photosynthetic yield, crucial for life to exist on Earth. Photosynthetic acclimation is an excellent example of the complexity of biological systems, in which highly diverse processes, ranging from electron excitation over protein protonation to enzymatic processes coupling ion gradients with biosynthetic activity interact on drastically different timescales, ranging from picoseconds to hours. An efficient functioning of the photosynthetic apparatus and its protection is paramount for efficient downstream processes including metabolism and growth. Modern experimental techniques can be successfully integrated with theoretical and mathematical models to promote our understanding of underlying mechanisms and principles. This Review aims to provide a retrospective analysis of multidisciplinary photosynthetic acclimation research carried out by members of the Marie Curie Initial Training Project “AccliPhot”, placing the results in a wider context. The Review also highlights the applicability of photosynthetic organisms for industry, particularly with regards to the cultivation of microalgae. It aims to demonstrate how theoretical concepts can successfully complement experimental studies broadening our knowledge of common principles in acclimation processes in photosynthetic organisms, as well as in the field of applied microalgal biotechnology

L'utilisation de la lumière chez les microalgues : la diatomée marine Phaeodactylum tricornutum et l'algue verte Chlamydomonas reinhardtii

Microalgae have developed distinct approaches to modulate light absorption and utilization by their photosystems in response to environmental stimuli. In this Ph.D Thesis, I characterised different strategies employed by freshwater (Chlamydomonas reinhardtii) and marine algae (Phaeodactylum tricornutum) to optimise their acclimation to the environment.In the first part of this work, I used spectroscopic, biochemical, electron microscopy analysis and 3-dimentional reconstitution to generate a model of the entire cell of the marine diatom Phaeodactylum tricornutum. This model has been used to address the following questions: i. how is a secondary chloroplast structured to facilitate exchanges with the cytosol via its four membranes envelope barrier ii. how have diatoms shaped their photosynthetic membranes to optimise light absorption and downstream electron flow and iii. how the cellular organelles interact to optimise CO2 assimilation via ATP/NADPH exchanges.In the second part, I have focused on the regulation of light harvesting and dissipation in Chlamydomonas by studying the role of perception of light colour and metabolism on excess light dissipation via the Non-Photochemical Quenching of energy (NPQ). Using biochemical and spectroscopic approaches, I found a molecular link between photoreception, photosynthesis and photoprotection in Chlamydomonas via the role of the photoreceptor phototropin on excess absorbed energy dissipation (NPQ) and also demonstrated that besides light, downstream metabolism can also affect this acclimation process.Overall this Ph.D work reveals the existence and integration of different signal pathways in the regulation of photoprotective responses by microalgae living in the ocean and in the land.Les microalgues ont développé des approches distinctes pour moduler l'absorption de la lumière et son utilisation par leurs photosystèmes en réponse à des stimuli environnementaux. Dans ce rapport de Thèse je présente les différentes stratégies employées par une algue d'eau douce (Chlamydomonas reinhardtii) et une algue marine (Phaeodactylum tricornutum) pour optimiser leur acclimatation à l'environnement.Dans la première partie de ce rapport, je propose un modèle de cellules entières de la diatomée marine Phaeodactylum tricornutum obtenue par analyses spectroscopiques et biochimiques ainsi que par l’obtention d’images par microscopie électronique et reconstitution 3-D. Ce modèle a été utilisé pour répondre aux questions suivantes i. comment est structuré un chloroplaste secondaire pour faciliter les échanges avec le cytosol à travers les quatre membranes qui le délimitent ii. comment sont structurées les membranes photosynthétiques afin d’optimiser l'absorption de lumière et le flux d'électrons et iii. comment les chloroplastes et les mitochondries sont organisés pour optimiser l'assimilation du CO2 par échange ATP / NADPH.La deuxième partie de ce rapport porte sur la régulation de la collection de la lumière et de sa dissipation chez Chlamydomonas grâce à l'étude d'une part du rôle de la perception de la couleur de la lumière et d'autre part du métabolisme sur la dissipation de l'excès de lumière par quenching non photochimique (NPQ). En utilisant des approches biochimiques et spectroscopiques, j'ai mis en évidence un lien moléculaire entre la photoréception, la photosynthèse et la photoprotection chez Chlamydomonas via le rôle du photorécepteur phototropine, démontrant ainsi que le métabolisme, en plus de la lumière, peut aussi affecter ce processus d'acclimatation.En conclusion, ce travail de thèse révèle l'existence et l'intégration des différentes voies de signalisation dans la régulation des réponses photoprotectrices mises en place chez les microalgues marines et d'eau douce

The Velocity of Light Intensity Increase Modulates the Photoprotective Response in Coastal Diatoms

International audienceIn aquatic ecosystems, the superimposition of mixing events to the light diel cycle exposes phytoplankton to changes in the velocity of light intensity increase, from diurnal variations to faster mixing-related ones. This is particularly true in coastal waters, where diatoms are dominant. This study aims to investigate if coastal diatoms differently activate the photoprotective responses, xanthophyll cycle (XC) and non-photochemical fluorescence quenching (NPQ), to cope with predictable light diel cycle and unpredictable mixing-related light variations. We compared the effect of two fast light intensity increases (simulating mixing events) with that of a slower increase (corresponding to the light diel cycle) on the modulation of XC and NPQ in the planktonic coastal diatom Pseudo-nitzschia multistriata. During each light treatment, the photon flux density (PFD) progressively increased from darkness to five peaks, ranging from 100 to 650 mmol photons m 22 s 21 . Our results show that the diel cycle-related PFD increase strongly activates XC through the enhancement of the carotenoid biosynthesis and induces a moderate and gradual NPQ formation over the light gradient. In contrast, during mixing-related PFD increases, XC is less activated, while higher NPQ rapidly develops at moderate PFD. We observe that together with the light intensity and its increase velocity, the saturation light for photosynthesis (Ek) is a key parameter in modulating photoprotection. We propose that the capacity to adequately regulate and actuate alternative photoprotective 'safety valves' in response to changing velocity of light intensity increase further enhances the photophysiological flexibility of diatoms. This might be an evolutionary outcome of diatom adaptation to turbulent marine ecosystems characterized by unpredictable mixing-related light changes over the light diel cycle

Ultrastructure of the Periplastidial Compartment of the Diatom Phaeodactylum tricornutum.

International audienceDiatoms contain a secondary plastid that derives from a red algal symbiont. This organelle is limited by four membranes. The two outermost membranes are the chloroplast endoplasmic reticulum membrane (cERM), which is continuous with the host outer nuclear envelope, and the periplastidial membrane (PPM). The two innermost membranes correspond to the outer and inner envelope membranes (oEM and iEM) of the symbiont's chloroplast. Between the PPM and oEM lies a minimized symbiont cytoplasm, the periplastidial compartment (PPC). In Phaeodactylum tricornutum, PPC-resident proteins are localized in "blob-like-structures", which remain associated with plastids after cell disruption. We analyzed disrupted Phaeodactylum cells by focused ion beam scanning electron microscopy, revealing the presence of a vesicular network (VN) in the PPC, at a location consistent with blob-like structures. Presence of a VN in the PPC was confirmed in intact cells. Additionally, direct membrane contacts were observed between the PPM and nuclear inner envelope membrane at the level of the chloroplast-nucleus isthmus. This study provides insights into the PPC ultrastructure and opens perspectives on the function of this residual cytoplasm of red algal origin

Photosynthetic and physiological properties, and photosynthetic pigment content of <i>Pseudo-nitzschia multistriata</i>.

<p>The measurement of photosynthetic and physiological properties was performed on cells in the exponential growth phase, during preacclimation, the day before the experiments started. The growth rate did not change during experiments. _relETR_max, maximal relative electron transport rate (in mol e⁻ g Chl <i>a</i>⁻¹ h⁻¹); Ek, saturation light for photosynthesis (in µmol photons m⁻² s⁻¹); µ, growth rate (in d⁻¹); F_v/F_m, photosystem II maximal photochemical efficiency. Values are means ± SD (<i>n</i> = 9). Chlorophyll <i>a</i> cellular content (Chl <i>a</i>, in 10⁻¹⁶ mol Chl <i>a</i> cell⁻¹) and photosynthetic accessory pigments Chl <i>a</i>⁻¹ content (in mol pigment/100 mol Chl <i>a</i>) measurements were performed during experiments. Fuco, fucoxanthin: Chl <i>c</i>, chlorophyll <i>c</i>₁,₂,₃. Pigment data are means ± SD of the all data set (<i>n</i> = 135).</p

Influence of the kinetics of light increase on the photoprotection modulation.

<p>(A) Evolution of the number of absorbed photons per Chl <i>a</i> integrated over time (integrated absorbed light, Int Abs Light; expressed in mol photons mg Chl <i>a</i>⁻¹) over the light gradient, at the PFD peaks of 100, 250, 350, 500 and 650 µmol photons m⁻² s⁻¹, during the 5 h (white dots), 3 h (black squares) and 2 h kinetics of light increase (black triangles). Induction of the sustained light-acclimated NPQ (NPQ_sl; B) and evolution of the de-epoxidation state (DES = Dt/[Dd+Dt]; C) <i>versus</i> Int Abs Light during the 5 h (white dots), 3 h (black squares) and 2 h kinetics of light increase (black triangles). Values are means ± SD (<i>n</i> = 3).</p

Ions channels/transporters and chloroplast regulation.

International audienceIons play fundamental roles in all living cells and their gradients are often essential to fuel transports, to regulate enzyme activities and to transduce energy within and between cells. Their homeostasis is therefore an essential component of the cell metabolism. Ions must be imported from the extracellular matrix to their final subcellular compartments. Among them, the chloroplast is a particularly interesting example because there, ions not only modulate enzyme activities, but also mediate ATP synthesis and actively participate in the building of the photosynthetic structures by promoting membrane-membrane interaction. In this review, we first provide a comprehensive view of the different machineries involved in ion trafficking and homeostasis in the chloroplast, and then discuss peculiar functions exerted by ions in the frame of photochemical conversion of absorbed light energy

Cell Biology of Organelles

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Preacclimation and experimental light conditions.

<p>(A) <i>Pseudo-nitzschia multistriata</i> cells were grown under a sinusoidal light regime set to peak at the PFD of 100 µmol photons m⁻² s⁻¹ (preacclimation light, PL; dashed line). After two weeks of preacclimation, cells in the exponential growth phase were shifted to three experimental light treatments, the 5 h (diel cycle-related PFD increase; B), 3 h and 2 h kinetics of light increase (mixing-related PFD increases; C and D, respectively), each characterized by light gradual increases peaking at the PFD of 100, 250, 350, 500 and 650 µmol photons m⁻² s⁻¹. In each panel, experimental light increases (solid lines) are compared to PL (dashed line). Triplicate samples were taken at three sampling time points during light increase (dots, B−D). Firstly, cultures were sampled in darkness. Then, after 3 h (5 h kinetics), 2 h (3 h kinetics), and 1.5 h (2 h kinetics), samples were taken at the PFD of 42, 123, 150, 164 and 280 µmol photons m⁻² s⁻¹ for the light condition peaking at 100, 250, 350, 500 and 650 µmol photons m⁻² s⁻¹, respectively. Lastly, cultures were sampled at PFD peaks.</p