125 results on '"Morosinotto T"'
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2. Crystal Structure of the Lipocalin domain of Violaxanthin de-epoxidase (VDE) at pH5
- Author
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Arnoux, P., primary, Morosinotto, T., additional, and Pignol, D., additional
- Published
- 2009
- Full Text
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3. An IdentifiableState Model To Describe Light IntensityInfluence on Microalgae Growth.
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Bernardi, A., Perin, G., Sforza, E., Galvanin, F., Morosinotto, T., and Bezzo, F.
- Published
- 2014
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4. Vegetal oil from microalgae: mixotrophic growth in view of large-scale production
- Author
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Sforza, E., Bertucco, A., Morosinotto, T., and Giacometti, G.M.
- Published
- 2010
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5. Antenna complexes protect Photosystem I from Photoinhibition
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Hienerwadel Rainer, Ballottari Matteo, Alboresi Alessandro, Giacometti Giorgio M, and Morosinotto Tomas
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Botany ,QK1-989 - Abstract
Abstract Background Photosystems are composed of two moieties, a reaction center and a peripheral antenna system. In photosynthetic eukaryotes the latter system is composed of proteins belonging to Lhc family. An increasing set of evidences demonstrated how these polypeptides play a relevant physiological function in both light harvesting and photoprotection. Despite the sequence similarity between antenna proteins associated with the two Photosystems, present knowledge on their physiological role is mostly limited to complexes associated to Photosystem II. Results In this work we analyzed the physiological role of Photosystem I antenna system in Arabidopsis thaliana both in vivo and in vitro. Plants depleted in individual antenna polypeptides showed a reduced capacity for photoprotection and an increased production of reactive oxygen species upon high light exposure. In vitro experiments on isolated complexes confirmed that depletion of antenna proteins reduced the resistance of isolated Photosystem I particles to high light and that the antenna is effective in photoprotection only upon the interaction with the core complex. Conclusion We show that antenna proteins play a dual role in Arabidopsis thaliana Photosystem I photoprotection: first, a Photosystem I with an intact antenna system is more resistant to high light because of a reduced production of reactive oxygen species and, second, antenna chlorophyll-proteins are the first target of high light damages. When photoprotection mechanisms become insufficient, the antenna chlorophyll proteins act as fuses: LHCI chlorophylls are degraded while the reaction center photochemical activity is maintained. Differences with respect to photoprotection strategy in Photosystem II, where the reaction center is the first target of photoinhibition, are discussed.
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- 2009
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6. Physcomitrium patens flavodiiron proteins form heterotetrametric complexes.
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Beraldo C, Traverso E, Boschin M, Cendron L, Morosinotto T, and Alboresi A
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- Photosystem I Protein Complex metabolism, Photosystem I Protein Complex chemistry, Protein Multimerization, Iron metabolism, Iron chemistry, Escherichia coli metabolism, Escherichia coli genetics, Bryopsida metabolism, Plant Proteins metabolism, Plant Proteins chemistry
- Abstract
Flavodiiron proteins (FLVs) catalyze the reduction of oxygen to water by using electrons from Photosystem I (PSI). In several photosynthetic organisms such as cyanobacteria, green algae, mosses and gymnosperms, FLV-dependent electron flow protects PSI from over-reduction and consequent damage especially under fluctuating light conditions. In this work we investigated biochemical and structural properties of FLVA and FLVB from the model moss Physcomitrium patens. The two proteins, expressed and purified from Escherichia coli, bind both iron and flavin cofactors and show NAD(P)H oxidase activity as well as oxygen reductase capacities. Moreover, the co-expression of both FLVA and FLVB, coupled to a tandem affinity purification procedure with two different affinity tags, enabled the isolation of the stable and catalytically active FLVA/B hetero tetrameric protein complex with cooperative nature. The multimeric organization was shown to be stabilized by inter-subunit disulfide bonds. This investigation provides valuable new information on the biochemical properties of FLVs, with new insights into their in vivo activity., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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7. Mitochondrial respiration is essential for photosynthesis-dependent ATP supply of the plant cytosol.
- Author
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Vera-Vives AM, Novel P, Zheng K, Tan SL, Schwarzländer M, Alboresi A, and Morosinotto T
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- Light, Adenosine Triphosphate metabolism, Photosynthesis, Cytosol metabolism, Mitochondria metabolism, Cell Respiration, Bryopsida metabolism, Bryopsida genetics, Bryopsida growth & development
- Abstract
Plants rely on solar energy to synthesize ATP and NADPH for photosynthetic carbon fixation and all cellular need. Mitochondrial respiration is essential in plants, but this may be due to heterotrophic bottlenecks during plant development or because it is also necessary in photosynthetically active cells. In this study, we examined in vivo changes of cytosolic ATP concentration in response to light, employing a biosensing strategy in the moss Physcomitrium patens and revealing increased cytosolic ATP concentration caused by photosynthetic activity. Plants depleted of respiratory Complex I showed decreased cytosolic ATP accumulation, highlighting a critical role of mitochondrial respiration in light-dependent ATP supply of the cytosol. Consistently, targeting mitochondrial ATP production directly, through the construction of mutants deficient in mitochondrial ATPase (complex V), led to drastic growth reduction, despite only minor alterations in photosynthetic electron transport activity. Since P. patens is photoautotrophic throughout its development, we conclude that heterotrophic bottlenecks cannot account for the indispensable role of mitochondrial respiration in plants. Instead, our results support that mitochondrial respiration is essential for ATP provision to the cytosol in photosynthesizing cells. Mitochondrial respiration provides metabolic integration, ensuring supply of cytosolic ATP essential for supporting plant growth and development., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)
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- 2024
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8. Autotrophic poly-3-hydroxybutyrate accumulation in Cupriavidus necator for sustainable bioplastic production triggered by nutrient starvation.
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Santin A, Spatola Rossi T, Morlino MS, Gupte AP, Favaro L, Morosinotto T, Treu L, and Campanaro S
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- Autotrophic Processes, Oxygen metabolism, Phosphorus metabolism, Carbon metabolism, Nutrients metabolism, Plastics metabolism, Hydrogen metabolism, Biodegradable Plastics metabolism, Magnesium metabolism, Polyhydroxybutyrates, Cupriavidus necator metabolism, Hydroxybutyrates metabolism, Polyesters metabolism, Nitrogen metabolism
- Abstract
Cupriavidus necator is a facultative chemolithoautotrophic bacterium able to convert carbon dioxide into poly-3-hydroxybutyrate. This is highly promising as the conversion process allows the production of sustainable and biodegradable plastics. Poly-3-hydroxybutyrate accumulation is known to be induced by nutrient starvation, but information regarding the optimal stress conditions controlling the process is still heterogeneous and fragmentary. This study presents a comprehensive comparison of the effects of nutrient stress conditions, namely nitrogen, hydrogen, phosphorus, oxygen, and magnesium deprivation, on poly-3-hydroxybutyrate accumulation in C. necator DSM545. Nitrogen starvation exhibited the highest poly-3-hydroxybutyrate accumulation, achieving 54% of total cell dry weight after four days of nutrient stress, and a carbon conversion efficiency of 85%. The gas consumption patterns indicated flexible physiological mechanisms underlying polymer accumulation and depolymerization. These findings provide insights into strategies for efficient carbon conversion into bioplastics, and highlight the key role of C. necator for future industrial-scale applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
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- 2024
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9. Deciphering the genetic landscape of enhanced poly-3-hydroxybutyrate production in Synechocystis sp. B12.
- Author
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Santin A, Collura F, Singh G, Morlino MS, Bizzotto E, Bellan A, Gupte AP, Favaro L, Campanaro S, Treu L, and Morosinotto T
- Abstract
Background: Microbial biopolymers such as poly-3-hydroxybutyrate (PHB) are emerging as promising alternatives for sustainable production of biodegradable bioplastics. Their promise is heightened by the potential utilisation of photosynthetic organisms, thus exploiting sunlight and carbon dioxide as source of energy and carbon, respectively. The cyanobacterium Synechocystis sp. B12 is an attractive candidate for its superior ability to accumulate high amounts of PHB as well as for its high-light tolerance, which makes it extremely suitable for large-scale cultivation. Beyond its practical applications, B12 serves as an intriguing model for unravelling the molecular mechanisms behind PHB accumulation., Results: Through a multifaceted approach, integrating physiological, genomic and transcriptomic analyses, this work identified genes involved in the upregulation of chlorophyll biosynthesis and phycobilisome degradation as the possible candidates providing Synechocystis sp. B12 an advantage in growth under high-light conditions. Gene expression differences in pentose phosphate pathway and acetyl-CoA metabolism were instead recognised as mainly responsible for the increased Synechocystis sp. B12 PHB production during nitrogen starvation. In both response to strong illumination and PHB accumulation, Synechocystis sp. B12 showed a metabolic modulation similar but more pronounced than the reference strain, yielding in better performances., Conclusions: Our findings shed light on the molecular mechanisms of PHB biosynthesis, providing valuable insights for optimising the use of Synechocystis in economically viable and sustainable PHB production. In addition, this work supplies crucial knowledge about the metabolic processes involved in production and accumulation of these molecules, which can be seminal for the application to other microorganisms as well., (© 2024. The Author(s).)
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- 2024
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10. Net O 2 exchange rates under dark and light conditions across different stem compartments.
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Natale S, Peralta Ogorek LL, Caracciolo L, Morosinotto T, van Amerongen H, Casolo V, Pedersen O, and Nardini A
- Subjects
- Darkness, Fraxinus metabolism, Chloroplasts metabolism, Chloroplasts radiation effects, Plant Bark metabolism, Photosynthesis radiation effects, Photosystem II Protein Complex metabolism, Plant Stems metabolism, Plant Stems radiation effects, Oxygen metabolism, Light, Wood metabolism
- Abstract
Woody plants display some photosynthetic activity in stems, but the biological role of stem photosynthesis and the specific contributions of bark and wood to carbon uptake and oxygen evolution remain poorly understood. We aimed to elucidate the functional characteristics of chloroplasts in stems of different ages in Fraxinus ornus. Our investigation employed diverse experimental approaches, including microsensor technology to assess oxygen production rates in whole stem, bark, and wood separately. Additionally, we utilized fluorescence lifetime imaging microscopy (FLIM) to characterize the relative abundance of photosystems I and II (PSI : PSII chlorophyll ratio) in bark and wood. Our findings revealed light-induced increases in O
2 production in whole stem, bark, and wood. We present the radial profile of O2 production in F. ornus stems, demonstrating the capability of stem chloroplasts to perform light-dependent electron transport. Younger stems exhibited higher light-induced O2 production and dark respiration rates than older ones. While bark emerged as the primary contributor to net O2 production under light conditions, our data underscored that wood chloroplasts are also photosynthetically active. The FLIM analysis unveiled a lower PSI abundance in wood than in bark, suggesting stem chloroplasts are not only active but also acclimate to the spectral composition of light reaching inner compartments., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)- Published
- 2024
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11. Biological carbon capture from biogas streams: Insights into Cupriavidus necator autotrophic growth and transcriptional profile.
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Serna-García R, Silvia Morlino M, Bucci L, Savio F, Favaro L, Morosinotto T, Seco A, Bouzas A, Campanaro S, and Treu L
- Subjects
- Carbon Dioxide, Biofuels, Rivers, Autotrophic Processes, Cupriavidus necator genetics, Polyhydroxyalkanoates metabolism
- Abstract
Recycling carbon-rich wastes into high-value platform chemicals through biological processes provides a sustainable alternative to petrochemicals. Cupriavidus necator, known for converting carbon dioxide (CO
2 ) into polyhydroxyalkanoates (PHA) was studied for the first time using biogas streams as the sole carbon source. The bacterium efficiently consumed biogenic CO2 from raw biogas with methane at high concentrations (50%) proving non-toxic. Continuous addition of H2 and O2 enabled growth trends comparable to glucose-based heterotrophic growth. Transcriptomic analysis revealed CO2 -adaptated cultures exhibited upregulation of hydrogenases and Calvin cycle enzymes, as well as genes related to electron transport, nutrient uptake, and glyoxylate cycle. Non-adapted samples displayed activation of stress response mechanisms, suggesting potential lags in large-scale processes. These findings showcase the setting of growth parameters for a pioneering biological biogas upgrading strategy, emphasizing the importance of inoculum adaptation for autotrophic growth and providing potential targets for genetic engineering to push PHA yields in future applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)- Published
- 2024
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12. Assessment of photosynthetic activity in dense microalgae cultures using oxygen production.
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Vera-Vives AM, Michelberger T, Morosinotto T, and Perin G
- Subjects
- Ecosystem, Oxygen metabolism, Photosynthesis, Photobioreactors, Biomass, Microalgae
- Abstract
Microalgae are photosynthetic microorganisms playing a pivotal role in primary production in aquatic ecosystems, sustaining the entry of carbon in the biosphere. Microalgae have also been recognized as sustainable source of biomass to complement crops. For this objective they are cultivated in photobioreactors or ponds at high cell density to maximize biomass productivity and lower the cost of downstream processes. Photosynthesis depends on light availability, that is often not constant over time. In nature, sunlight fluctuates over diurnal cycles and weather conditions. In high-density microalgae cultures of photobioreactors outdoors, on top of natural variations, microalgae are subjected to further complexity in light exposure. Because of the high-density cells experience self-shading effects that heavily limit light availability in most of the mass culture volume. This limitation strongly affects biomass productivity of industrial microalgae cultivation plants with important implications on economic feasibility. Understanding how photosynthesis responds to cell density is informative to assess functionality in the inhomogeneous light environment of industrial photobioreactors. In this work we exploited a high-sensitivity Clark electrode to measure microalgae photosynthesis and compare cultures with different densities, using Nannochloropsis as model organism. We observed that cell density has a substantial impact on photosynthetic activity, and demonstrated the reduction of the cell's light-absorption capacity by genetic modification is a valuable strategy to increase photosynthetic functionality on a chlorophyll-basis of dense microalgae cultures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.)
- Published
- 2024
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13. Cupriavidus necator as a platform for polyhydroxyalkanoate production: An overview of strains, metabolism, and modeling approaches.
- Author
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Morlino MS, Serna García R, Savio F, Zampieri G, Morosinotto T, Treu L, and Campanaro S
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- Fermentation, Biotechnology, Carbon metabolism, Polyhydroxyalkanoates genetics, Polyhydroxyalkanoates metabolism, Cupriavidus necator genetics, Cupriavidus necator metabolism
- Abstract
Cupriavidus necator is a bacterium with a high phenotypic diversity and versatile metabolic capabilities. It has been extensively studied as a model hydrogen oxidizer, as well as a producer of polyhydroxyalkanoates (PHA), plastic-like biopolymers with a high potential to substitute petroleum-based materials. Thanks to its adaptability to diverse metabolic lifestyles and to the ability to accumulate large amounts of PHA, C. necator is employed in many biotechnological processes, with particular focus on PHA production from waste carbon sources. The large availability of genomic information has enabled a characterization of C. necator's metabolism, leading to the establishment of metabolic models which are used to devise and optimize culture conditions and genetic engineering approaches. In this work, the characteristics of available C. necator strains and genomes are reviewed, underlining how a thorough comprehension of the genetic variability of C. necator is lacking and it could be instrumental for wider application of this microorganism. The metabolic paradigms of C. necator and how they are connected to PHA production and accumulation are described, also recapitulating the variety of carbon substrates used for PHA accumulation, highlighting the most promising strategies to increase the yield. Finally, the review describes and critically analyzes currently available genome-scale metabolic models and reduced metabolic network applications commonly employed in the optimization of PHA production. Overall, it appears that the capacity of C. necator of performing CO
2 bioconversion to PHA is still underexplored, both in biotechnological applications and in metabolic modeling. However, the accurate characterization of this organism and the efforts in using it for gas fermentation can help tackle this challenging perspective in the future., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2023
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14. Improving photosynthetic efficiency toward food security: Strategies, advances, and perspectives.
- Author
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Smith EN, van Aalst M, Tosens T, Niinemets Ü, Stich B, Morosinotto T, Alboresi A, Erb TJ, Gómez-Coronado PA, Tolleter D, Finazzi G, Curien G, Heinemann M, Ebenhöh O, Hibberd JM, Schlüter U, Sun T, and Weber APM
- Subjects
- Crops, Agricultural, Nutrients, Food Security, Plant Breeding, Photosynthesis
- Abstract
Photosynthesis in crops and natural vegetation allows light energy to be converted into chemical energy and thus forms the foundation for almost all terrestrial trophic networks on Earth. The efficiency of photosynthetic energy conversion plays a crucial role in determining the portion of incident solar radiation that can be used to generate plant biomass throughout a growth season. Consequently, alongside the factors such as resource availability, crop management, crop selection, maintenance costs, and intrinsic yield potential, photosynthetic energy use efficiency significantly influences crop yield. Photosynthetic efficiency is relevant to sustainability and food security because it affects water use efficiency, nutrient use efficiency, and land use efficiency. This review focuses specifically on the potential for improvements in photosynthetic efficiency to drive a sustainable increase in crop yields. We discuss bypassing photorespiration, enhancing light use efficiency, harnessing natural variation in photosynthetic parameters for breeding purposes, and adopting new-to-nature approaches that show promise for achieving unprecedented gains in photosynthetic efficiency., (Copyright © 2023 The Author. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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15. Modulation of xanthophyll cycle impacts biomass productivity in the marine microalga Nannochloropsis .
- Author
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Perin G, Bellan A, Michelberger T, Lyska D, Wakao S, Niyogi KK, and Morosinotto T
- Subjects
- Biomass, Zeaxanthins, Xanthophylls, Microalgae
- Abstract
Life on earth depends on photosynthetic primary producers that exploit sunlight to fix CO
2 into biomass. Approximately half of global primary production is associated with microalgae living in aquatic environments. Microalgae also represent a promising source of biomass to complement crop cultivation, and they could contribute to the development of a more sustainable bioeconomy. Photosynthetic organisms evolved multiple mechanisms involved in the regulation of photosynthesis to respond to highly variable environmental conditions. While essential to avoid photodamage, regulation of photosynthesis results in dissipation of absorbed light energy, generating a complex trade-off between protection from stress and light-use efficiency. This work investigates the impact of the xanthophyll cycle, the light-induced reversible conversion of violaxanthin into zeaxanthin, on the protection from excess light and on biomass productivity in the marine microalgae of the genus Nannochloropsis. Zeaxanthin is shown to have an essential role in protection from excess light, contributing to the induction of nonphotochemical quenching and scavenging of reactive oxygen species. On the contrary, the overexpression of zeaxanthin epoxidase enables a faster reconversion of zeaxanthin to violaxanthin that is shown to be advantageous for biomass productivity in dense cultures in photobioreactors. These results demonstrate that zeaxanthin accumulation is critical to respond to strong illumination, but it may lead to unnecessary energy losses in light-limiting conditions and accelerating its reconversion to violaxanthin provides an advantage for biomass productivity in microalgae.- Published
- 2023
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16. Structure and function of bark and wood chloroplasts in a drought-tolerant tree (Fraxinus ornus L.).
- Author
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Natale S, La Rocca N, Battistuzzi M, Morosinotto T, Nardini A, and Alboresi A
- Subjects
- Wood metabolism, Chlorophyll A metabolism, Droughts, Plant Bark metabolism, Chloroplasts metabolism, Photosynthesis, Chlorophyll metabolism, Light, Plant Leaves metabolism, Photosystem II Protein Complex, Trees metabolism, Fraxinus
- Abstract
Leaves are the most important photosynthetic organs in most woody plants, but chloroplasts are also found in organs optimized for other functions. However, the actual photosynthetic efficiency of these chloroplasts is still unclear. We analyzed bark and wood chloroplasts of Fraxinus ornus L. saplings. Optical and spectroscopic methods were applied to stem samples and compared with leaves. A sharp light gradient was detected along the stem radial direction, with blue light mainly absorbed by the outer bark, and far-red-enriched light reaching the underlying xylem and pith. Chlorophylls were evident in the xylem rays and the pith and showed an increasing concentration gradient toward the bark. The stem photosynthetic apparatus showed features typical of acclimation to a low-light environment, such as larger grana stacks, lower chlorophyll a/b and photosystem I/II ratios compared with leaves. Despite likely receiving very few photons, wood chloroplasts were photosynthetically active and fully capable of generating a light-dependent electron transport. Our data provide a comprehensive scenario of the functional features of bark and wood chloroplasts in a woody species and suggest that stem photosynthesis is coherently optimized to the prevailing micro-environmental conditions at the bark and wood level., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)
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- 2023
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17. Functional analysis of PsbS transmembrane domains through base editing in Physcomitrium patens.
- Author
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Beraldo C, Guyon-Debast A, Alboresi A, Nogué F, and Morosinotto T
- Subjects
- Photosystem II Protein Complex metabolism, Gene Editing, Light-Harvesting Protein Complexes metabolism, Photosynthesis, Light
- Abstract
Plants exposed to light fluctuations are protected from photodamage by non-photochemical quenching (NPQ), a reversible mechanism that enables dissipation of excess absorbed energy as heat, which is essential for plant fitness and crop productivity. In plants NPQ requires the presence of the membrane protein PsbS, which upon activation interacts with antenna proteins, inducing their dissipative conformation. Here, we exploited base editing (BE) in the moss Physcomitrium patens to introduce specific amino acid changes in vivo and assess their impact on PsbS activity, targeting transmembrane regions to investigate their role in essential protein-protein interactions. This approach enabled the recognition of residues essential for protein stability and the identification of a hydrophobic cluster of amino acids impacting PsbS activity. This work provides new information on the molecular mechanism of PsbS while also demonstrating the potential of BE approaches for in planta gene function analysis., (© 2023 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)
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- 2023
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18. Oxygenic photosynthetic responses of cyanobacteria exposed under an M-dwarf starlight simulator: Implications for exoplanet's habitability.
- Author
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Battistuzzi M, Cocola L, Claudi R, Pozzer AC, Segalla A, Simionato D, Morosinotto T, Poletto L, and La Rocca N
- Abstract
Introduction: The search for life on distant exoplanets is expected to rely on atmospheric biosignatures detection, such as oxygen of biological origin. However, it is not demonstrated how much oxygenic photosynthesis, which on Earth depends on visible light, could work under spectral conditions simulating exoplanets orbiting the Habitable Zone of M-dwarf stars, which have low light emission in the visible and high light emission in the far-red/near-infrared. By utilizing cyanobacteria, the first organisms to evolve oxygenic photosynthesis on our planet, and a starlight simulator capable of accurately reproducing the emission spectrum of an M-dwarf in the range 350-900 nm, we could answer this question., Methods: We performed experiments with the cyanobacterium Chlorogloeopsis fritschii PCC6912, capable of Far-Red Light Photoacclimation (FaRLiP), which allows the strain to harvest far-red in addition to visible light for photosynthesis, and Synechocystis sp. PCC6803, a species unable to perform this photoacclimation, comparing their responses when exposed to three simulated light spectra: M-dwarf, solar and far-red. We analysed growth and photosynthetic acclimation features in terms of pigment composition and photosystems organization. Finally, we determined the oxygen production of the strains directly exposed to the different spectra., Results: Both cyanobacteria were shown to grow and photosynthesize similarly under M-dwarf and solar light conditions: Synechocystis sp. by utilizing the few photons in the visible, C. fritschii by harvesting both visible and far-red light, activating the FaRLiP response., Discussion: Our results experimentally show that an M-dwarf light spectrum could support a biological oxygen production similar to that in solar light at the tested light intensities, suggesting the possibility to discover such atmospheric biosignatures on those exoplanets if other boundary conditions are met., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Battistuzzi, Cocola, Claudi, Pozzer, Segalla, Simionato, Morosinotto, Poletto and La Rocca.)
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- 2023
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19. An increase in the membrane lipids recycling by PDAT overexpression stimulates the accumulation of triacylglycerol in Nannochloropsis gaditana.
- Author
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Fattore N, Bucci F, Bellan A, Bossi S, Maffei ME, and Morosinotto T
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- Membrane Lipids metabolism, Nitrogen metabolism, Plants metabolism, Triglycerides metabolism, Microalgae genetics, Microalgae metabolism, Stramenopiles genetics, Stramenopiles metabolism
- Abstract
Oleaginous microalgae represent potential feedstocks for the sustainable production of lipids thanks to their ability to accumulate triacylglycerols (TAGs). TAG accumulation in several algal species is strongly induced under specific conditions such as nutrient deprivation and high light which, however, also negatively impact growth. Genetic modification of lipogenic pathways can potentially enhance TAG accumulation without negatively affecting growth, avoiding the trade-off between biomass and lipid productivity. In this study, the phospholipid: diacylglycerol acyltransferase (PDAT), an enzyme involved in membrane lipid recycling, was overexpressed in the seawater alga Nannochloropsis gaditana. PDAT overexpression induced increased TAG content in actively growing algae cultures while no effects were observed in conditions naturally stimulating strong lipid accumulation such as high light and nitrogen starvation. The increase of TAG content was confirmed also in a strain cultivated in industrially relevant conditions even though PDAT overexpression, if too strong, the gene overexpression becomes detrimental for growth in the longer term. Results overall suggest that genetic modulation of the PDAT gene represents a promising strategy to increase microalgae lipid content by minimizing negative effects on biomass productivity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)
- Published
- 2022
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20. Role of serine/threonine protein kinase STN7 in the formation of two distinct photosystem I supercomplexes in Physcomitrium patens.
- Author
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Gerotto C, Trotta A, Bajwa AA, Morosinotto T, and Aro EM
- Subjects
- Light, Light-Harvesting Protein Complexes genetics, Light-Harvesting Protein Complexes metabolism, Phosphorylation, Photosystem I Protein Complex metabolism, Photosystem II Protein Complex metabolism, Protein Serine-Threonine Kinases, Serine metabolism, Threonine metabolism, Arabidopsis metabolism, Arabidopsis Proteins metabolism, Bryopsida genetics, Bryopsida metabolism
- Abstract
Reversible thylakoid protein phosphorylation provides most flowering plants with dynamic acclimation to short-term changes in environmental light conditions. Here, through generating Serine/Threonine protein kinase 7 (STN7)-depleted mutants in the moss Physcomitrella (Physcomitrium patens), we identified phosphorylation targets of STN7 kinase and their roles in short- and long-term acclimation of the moss to changing light conditions. Biochemical and mass spectrometry analyses revealed STN7-dependent phosphorylation of N-terminal Thr in specific Light-Harvesting Complex II (LHCII) trimer subunits (LHCBM2 and LHCBM4/8) and provided evidence that phospho-LHCBM accumulation is responsible for the assembly of two distinct Photosystem I (PSI) supercomplexes (SCs), both of which are largely absent in STN7-depleted mutants. Besides the canonical state transition complex (PSI-LHCI-LHCII), we isolated the larger moss-specific PSI-Large (PSI-LHCI-LHCB9-LHCII) from stroma-exposed thylakoids. Unlike PSI-LHCI-LHCII, PSI-Large did not demonstrate short-term dynamics for balancing the distribution of excitation energy between PSII and PSI. Instead, PSI-Large contributed to a more stable increase in PSI antenna size in Physcomitrella, except under prolonged high irradiance. Additionally, the STN7-depleted mutants revealed altered light-dependent phosphorylation of a monomeric antenna protein, LHCB6, whose phosphorylation displayed a complex regulation by multiple kinases. Collectively, the unique phosphorylation plasticity and dynamics of Physcomitrella monomeric LHCB6 and trimeric LHCBM isoforms, together with the presence of PSI SCs with different antenna sizes and responsiveness to light changes, reflect the evolutionary position of mosses between green algae and vascular plants, yet with clear moss-specific features emphasizing their adaptation to terrestrial low-light environments., (© The Author(s) 2022. Published by Oxford University Press on behalf of American Society of Plant Biologists.)
- Published
- 2022
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21. Knowledge of Regulation of Photosynthesis in Outdoor Microalgae Cultures Is Essential for the Optimization of Biomass Productivity.
- Author
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Perin G, Gambaro F, and Morosinotto T
- Abstract
Microalgae represent a sustainable source of biomass that can be exploited for pharmaceutical, nutraceutical, cosmetic applications, as well as for food, feed, chemicals, and energy. To make microalgae applications economically competitive and maximize their positive environmental impact, it is however necessary to optimize productivity when cultivated at a large scale. Independently from the final product, this objective requires the optimization of biomass productivity and thus of microalgae ability to exploit light for CO
2 fixation. Light is a highly variable environmental parameter, continuously changing depending on seasons, time of the day, and weather conditions. In microalgae large scale cultures, cell self-shading causes inhomogeneity in light distribution and, because of mixing, cells move between different parts of the culture, experiencing abrupt changes in light exposure. Microalgae evolved multiple regulatory mechanisms to deal with dynamic light conditions that, however, are not adapted to respond to the complex mixture of natural and artificial fluctuations found in large-scale cultures, which can thus drive to oversaturation of the photosynthetic machinery, leading to consequent oxidative stress. In this work, the present knowledge on the regulation of photosynthesis and its implications for the maximization of microalgae biomass productivity are discussed. Fast mechanisms of regulations, such as Non-Photochemical-Quenching and cyclic electron flow, are seminal to respond to sudden fluctuations of light intensity. However, they are less effective especially in the 1-100 s time range, where light fluctuations were shown to have the strongest negative impact on biomass productivity. On the longer term, microalgae modulate the composition and activity of the photosynthetic apparatus to environmental conditions, an acclimation response activated also in cultures outdoors. While regulation of photosynthesis has been investigated mainly in controlled lab-scale conditions so far, these mechanisms are highly impactful also in cultures outdoors, suggesting that the integration of detailed knowledge from microalgae large-scale cultivation is essential to drive more effective efforts to optimize biomass productivity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Perin, Gambaro and Morosinotto.)- Published
- 2022
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22. Protein dynamics and lipid affinity of monomeric, zeaxanthin-binding LHCII in thylakoid membranes.
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Azadi-Chegeni F, Thallmair S, Ward ME, Perin G, Marrink SJ, Baldus M, Morosinotto T, and Pandit A
- Subjects
- Photosynthesis, Photosystem II Protein Complex chemistry, Proteins metabolism, Zeaxanthins metabolism, Light-Harvesting Protein Complexes chemistry, Thylakoids metabolism
- Abstract
The xanthophyll cycle in the antenna of photosynthetic organisms under light stress is one of the most well-known processes in photosynthesis, but its role is not well understood. In the xanthophyll cycle, violaxanthin (Vio) is reversibly transformed to zeaxanthin (Zea) that occupies Vio binding sites of light-harvesting antenna proteins. Higher monomer/trimer ratios of the most abundant light-harvesting protein, the light-harvesting complex II (LHCII), usually occur in Zea accumulating membranes and have been observed in plants after prolonged illumination and during high-light acclimation. We present a combined NMR and coarse-grained simulation study on monomeric LHCII from the npq2 mutant that constitutively binds Zea in the Vio binding pocket. LHCII was isolated from
13 C-enriched npq2 Chlamydomonas reinhardtii (Cr) cells and reconstituted in thylakoid lipid membranes. NMR results reveal selective changes in the fold and dynamics of npq2 LHCII compared with the trimeric, wild-type and show that npq2 LHCII contains multiple mono- or digalactosyl diacylglycerol lipids (MGDG and DGDG) that are strongly protein bound. Coarse-grained simulations on npq2 LHCII embedded in a thylakoid lipid membrane agree with these observations. The simulations show that LHCII monomers have more extensive lipid contacts than LHCII trimers and that protein-lipid contacts are influenced by Zea. We propose that both monomerization and Zea binding could have a functional role in modulating membrane fluidity and influence the aggregation and conformational dynamics of LHCII with a likely impact on photoprotection ability., (Copyright © 2021 Biophysical Society. Published by Elsevier Inc. All rights reserved.)- Published
- 2022
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23. Acclimation of photosynthetic apparatus in the mesophilic red alga Dixoniella giordanoi.
- Author
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Fattore N, Savio S, Vera-Vives AM, Battistuzzi M, Moro I, La Rocca N, and Morosinotto T
- Subjects
- Acclimatization, Chlorophyll, Light, Photosynthesis, Photosystem II Protein Complex metabolism, Rhodophyta metabolism
- Abstract
Eukaryotic algae are photosynthetic organisms capable of exploiting sunlight to fix carbon dioxide into biomass with highly variable genetic and metabolic features. Information on algae metabolism from different species is inhomogeneous and, while green algae are, in general, more characterized, information on red algae is relatively scarce despite their relevant position in eukaryotic algae diversity. Within red algae, the best-known species are extremophiles or multicellular, while information on mesophilic unicellular organisms is still lacunose. Here, we investigate the photosynthetic properties of a recently isolated seawater unicellular mesophilic red alga, Dixoniella giordanoi. Upon exposure to different illuminations, D. giordanoi shows the ability to acclimate, modulate chlorophyll content, and re-organize thylakoid membranes. Phycobilisome content is also largely regulated, leading to almost complete disassembly of this antenna system in cells grown under intense illumination. Despite the absence of a light-induced xanthophyll cycle, cells accumulate zeaxanthin upon prolonged exposure to strong light, likely contributing to photoprotection. D. giordanoi cells show the ability to perform cyclic electron transport that is enhanced under strong illumination, likely contributing to the protection of Photosystem I from over-reduction and enabling cells to survive PSII photoinhibition without negative impact on growth., (© 2021 The Authors. Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)
- Published
- 2021
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24. Inactivation of mitochondrial complex I stimulates chloroplast ATPase in Physcomitrium patens.
- Author
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Mellon M, Storti M, Vera-Vives AM, Kramer DM, Alboresi A, and Morosinotto T
- Subjects
- Adenosine Triphosphatases metabolism, Bryopsida enzymology, Chloroplast Proteins metabolism, Plant Proteins metabolism, Adenosine Triphosphatases genetics, Bryopsida genetics, Chloroplast Proteins genetics, Plant Proteins genetics
- Abstract
Light is the ultimate source of energy for photosynthetic organisms, but respiration is fundamental for supporting metabolism during the night or in heterotrophic tissues. In this work, we isolated Physcomitrella (Physcomitrium patens) plants with altered respiration by inactivating Complex I (CI) of the mitochondrial electron transport chain by independently targeting on two essential subunits. Inactivation of CI caused a strong growth impairment even in fully autotrophic conditions in tissues where all cells are photosynthetically active, demonstrating that respiration is essential for photosynthesis. CI mutants showed alterations in the stoichiometry of respiratory complexes while the composition of photosynthetic apparatus was substantially unaffected. CI mutants showed altered photosynthesis with high activity of both Photosystems I and II, likely the result of high chloroplast ATPase activity that led to smaller ΔpH formation across thylakoid membranes, decreasing photosynthetic control on cytochrome b6f in CI mutants. These results demonstrate that alteration of respiratory activity directly impacts photosynthesis in P. patens and that metabolic interaction between organelles is essential in their ability to use light energy for growth., (© American Society of Plant Biologists 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2021
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25. Lipid Polymorphism of the Subchloroplast-Granum and Stroma Thylakoid Membrane-Particles. II. Structure and Functions.
- Author
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Dlouhý O, Karlický V, Arshad R, Zsiros O, Domonkos I, Kurasová I, Wacha AF, Morosinotto T, Bóta A, Kouřil R, Špunda V, and Garab G
- Subjects
- Circular Dichroism methods, Magnetic Resonance Spectroscopy methods, Microscopy, Electron methods, Photosynthesis genetics, Lipids genetics, Thylakoids genetics
- Abstract
In Part I, by using
31 P-NMR spectroscopy, we have shown that isolated granum and stroma thylakoid membranes (TMs), in addition to the bilayer, display two isotropic phases and an inverted hexagonal (HII ) phase; saturation transfer experiments and selective effects of lipase and thermal treatments have shown that these phases arise from distinct, yet interconnectable structural entities. To obtain information on the functional roles and origin of the different lipid phases, here we performed spectroscopic measurements and inspected the ultrastructure of these TM fragments. Circular dichroism, 77 K fluorescence emission spectroscopy, and variable chlorophyll-a fluorescence measurements revealed only minor lipase- or thermally induced changes in the photosynthetic machinery. Electrochromic absorbance transients showed that the TM fragments were re-sealed, and the vesicles largely retained their impermeabilities after lipase treatments-in line with the low susceptibility of the bilayer against the same treatment, as reflected by our31 P-NMR spectroscopy. Signatures of HII -phase could not be discerned with small-angle X-ray scattering-but traces of HII structures, without long-range order, were found by freeze-fracture electron microscopy (FF-EM) and cryo-electron tomography (CET). EM and CET images also revealed the presence of small vesicles and fusion of membrane particles, which might account for one of the isotropic phases. Interaction of VDE (violaxanthin de-epoxidase, detected by Western blot technique in both membrane fragments) with TM lipids might account for the other isotropic phase. In general, non-bilayer lipids are proposed to play role in the self-assembly of the highly organized yet dynamic TM network in chloroplasts.- Published
- 2021
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26. A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens.
- Author
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Guyon-Debast A, Alboresi A, Terret Z, Charlot F, Berthier F, Vendrell-Mir P, Casacuberta JM, Veillet F, Morosinotto T, Gallois JL, and Nogué F
- Subjects
- CRISPR-Cas Systems genetics, Clustered Regularly Interspaced Short Palindromic Repeats, Gene Editing, Mutagenesis, Site-Directed, Bryopsida genetics
- Abstract
CRISPR-Cas9 has proven to be highly valuable for genome editing in plants, including the model plant Physcomitrium patens. However, the fact that most of the editing events produced using the native Cas9 nuclease correspond to small insertions and deletions is a limitation. CRISPR-Cas9 base editors enable targeted mutation of single nucleotides in eukaryotic genomes and therefore overcome this limitation. Here, we report two programmable base-editing systems to induce precise cytosine or adenine conversions in P. patens. Using cytosine or adenine base editors, site-specific single-base mutations can be achieved with an efficiency up to 55%, without off-target mutations. Using the APT gene as a reporter of editing, we could show that both base editors can be used in simplex or multiplex, allowing for the production of protein variants with multiple amino-acid changes. Finally, we set up a co-editing selection system, named selecting modification of APRT to report gene targeting (SMART), allowing up to 90% efficiency site-specific base editing in P. patens. These two base editors will facilitate gene functional analysis in P. patens, allowing for site-specific editing of a given base through single sgRNA base editing or for in planta evolution of a given gene through the production of randomly mutagenised variants using multiple sgRNA base editing., (© 2021 The Authors New Phytologist © 2021 New Phytologist Foundation.)
- Published
- 2021
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27. Role of an ancient light-harvesting protein of PSI in light absorption and photoprotection.
- Author
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Lu Y, Gan Q, Iwai M, Alboresi A, Burlacot A, Dautermann O, Takahashi H, Crisanto T, Peltier G, Morosinotto T, Melis A, and Niyogi KK
- Subjects
- Adaptation, Physiological radiation effects, Chlorophyll A metabolism, Chlorophyll Binding Proteins genetics, Chlorophyll Binding Proteins isolation & purification, Mutation, Photosynthesis genetics, Photosynthesis radiation effects, Photosystem I Protein Complex genetics, Stramenopiles radiation effects, Adaptation, Physiological genetics, Chlorophyll Binding Proteins metabolism, Light adverse effects, Photosystem I Protein Complex metabolism, Stramenopiles physiology
- Abstract
Diverse algae of the red lineage possess chlorophyll a-binding proteins termed LHCR, comprising the PSI light-harvesting system, which represent an ancient antenna form that evolved in red algae and was acquired through secondary endosymbiosis. However, the function and regulation of LHCR complexes remain obscure. Here we describe isolation of a Nannochloropsis oceanica LHCR mutant, named hlr1, which exhibits a greater tolerance to high-light (HL) stress compared to the wild type. We show that increased tolerance to HL of the mutant can be attributed to alterations in PSI, making it less prone to ROS production, thereby limiting oxidative damage and favoring growth in HL. HLR1 deficiency attenuates PSI light-harvesting capacity and growth of the mutant under light-limiting conditions. We conclude that HLR1, a member of a conserved and broadly distributed clade of LHCR proteins, plays a pivotal role in a dynamic balancing act between photoprotection and efficient light harvesting for photosynthesis.
- Published
- 2021
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28. Conformational Dynamics of Light-Harvesting Complex II in a Native Membrane Environment.
- Author
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Azadi-Chegeni F, Ward ME, Perin G, Simionato D, Morosinotto T, Baldus M, and Pandit A
- Subjects
- Chlorophyll, Light-Harvesting Protein Complexes metabolism, Photosynthesis, Photosystem II Protein Complex metabolism, Chlamydomonas reinhardtii metabolism, Thylakoids metabolism
- Abstract
Photosynthetic light-harvesting complexes (LHCs) of higher plants, moss, and green algae can undergo dynamic conformational transitions, which have been correlated to their ability to adapt to fluctuations in the light environment. Herein, we demonstrate the application of solid-state NMR spectroscopy on native, heterogeneous thylakoid membranes of Chlamydomonas reinhardtii (Cr) and on Cr light-harvesting complex II (LHCII) in thylakoid lipid bilayers to detect LHCII conformational dynamics in its native membrane environment. We show that membrane-reconstituted LHCII contains selective sites that undergo fast, large-amplitude motions, including the phytol tails of two chlorophylls. Protein plasticity is also observed in the N-terminal stromal loop and in protein fragments facing the lumen, involving sites that stabilize the xanthophyll-cycle carotenoid violaxanthin and the two luteins. The results report on the intrinsic flexibility of LHCII pigment-protein complexes in a membrane environment, revealing putative sites for conformational switching. In thylakoid membranes, fast dynamics of protein and pigment sites is significantly reduced, which suggests that in their native organelle membranes, LHCII complexes are locked in specific conformational states., (Copyright © 2020 Biophysical Society. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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29. Light excess stimulates Poly-beta-hydroxybutyrate yield in a mangrove-isolated strain of Synechocystis sp.
- Author
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Gracioso LH, Bellan A, Karolski B, Cardoso LOB, Perpetuo EA, Nascimento CAOD, Giudici R, Pizzocchero V, Basaglia M, and Morosinotto T
- Subjects
- Carbon, Hydroxybutyrates, Polyesters, Synechocystis
- Abstract
Poly-β-hydroxybutyrate (PHB) is a biodegradable biopolymer that may replace fossil-based plastics reducing its negative environmental impact. One highly sustainable strategy to produce these biopolymers is the exploitation of photosynthetic microorganisms that use sunlight and CO
2 to produce biomass and subsequently, PHB. Exploring environmental biological diversity is a powerful tool to find resilient microorganisms potentially exploitable to produce bioproducts. In this work, a cyanobacterium (Synechocystis sp.) isolated from a contaminated area close to an important industrial complex was shown to produce PHB under different culture conditions. Carbon, nutrients supply and light intensity impact on biomass and PHB productivity were assessed, showing that the highest yield of PHB achieved was 241 mg L-1 (31%dcw ) under high light intensity. Remarkably this condition not only stimulated PHB accumulation by 70% compared to other conditions tested but also high cellular duplication rate, maximizing the potential of this strain for PHB production., (Copyright © 2020 Elsevier Ltd. All rights reserved.)- Published
- 2021
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30. Regulation of electron transport is essential for photosystem I stability and plant growth.
- Author
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Storti M, Segalla A, Mellon M, Alboresi A, and Morosinotto T
- Subjects
- Electron Transport, Light, Photosynthesis, Plant Development, Bryopsida metabolism, Photosystem I Protein Complex metabolism
- Abstract
Photosynthetic electron transport is regulated by cyclic and pseudocyclic electron flow (CEF and PCEF) to maintain the balance between light availability and metabolic demands. CEF transfers electrons from photosystem I to the plastoquinone pool with two mechanisms, dependent either on PGR5/PGRL1 or on the type I NADH dehydrogenase-like (NDH) complex. PCEF uses electrons from photosystem I to reduce oxygen and in many groups of photosynthetic organisms, but remarkably not in angiosperms, it is catalyzed by flavodiiron proteins (FLVs). In this study, Physcomitrella patens plants depleted in PGRL1, NDH and FLVs in different combinations were generated and characterized, showing that all these mechanisms are active in this moss. Surprisingly, in contrast to flowering plants, Physcomitrella patens can cope with the simultaneous inactivation of PGR5- and NDH-dependent CEF but, when FLVs are also depleted, plants show strong growth reduction and photosynthetic activity is drastically reduced. The results demonstrate that mechanisms for modulation of photosynthetic electron transport have large functional overlap but are together indispensable to protect photosystem I from damage and they are an essential component for photosynthesis in any light regime., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Foundation.)
- Published
- 2020
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31. The chloroplast NADH dehydrogenase-like complex influences the photosynthetic activity of the moss Physcomitrella patens.
- Author
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Storti M, Puggioni MP, Segalla A, Morosinotto T, and Alboresi A
- Subjects
- Chloroplasts metabolism, Electron Transport, Light, NADH Dehydrogenase genetics, NADH Dehydrogenase metabolism, Photosynthesis, Photosystem I Protein Complex metabolism, Bryopsida genetics, Bryopsida metabolism
- Abstract
Alternative electron pathways contribute to regulation of photosynthetic light reactions to adjust to metabolic demands in dynamic environments. The chloroplast NADH dehydrogenase-like (NDH) complex mediates the cyclic electron transport pathway around PSI in different cyanobacteria, algae, and plant species, but it is not fully conserved in all photosynthetic organisms. In order to assess how the physiological role of this complex changed during plant evolution, we isolated Physcomitrella patens lines knocked out for the NDHM gene that encodes a subunit fundamental for the activity of the complex. ndhm knockout mosses indicated high PSI acceptor side limitation upon abrupt changes in illumination. In P. patens, pseudo-cyclic electron transport mediated by flavodiiron proteins (FLVs) was also shown to prevent PSI over-reduction in plants exposed to light fluctuations. flva ndhm double knockout mosses had altered photosynthetic performance and growth defects under fluctuating light compared with the wild type and single knockout mutants. The results showed that while the contribution of NDH to electron transport is minor compared with FLV, NDH still participates in modulating photosynthetic activity, and it is critical to avoid PSI photoinhibition, especially when FLVs are inactive. The functional overlap between NDH- and FLV-dependent electron transport supports PSI activity and prevents its photoinhibition under light variations., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
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32. Higher order photoprotection mutants reveal the importance of ΔpH-dependent photosynthesis-control in preventing light induced damage to both photosystem II and photosystem I.
- Author
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Barbato R, Tadini L, Cannata R, Peracchio C, Jeran N, Alboresi A, Morosinotto T, Bajwa AA, Paakkarinen V, Suorsa M, Aro EM, and Pesaresi P
- Subjects
- Arabidopsis genetics, Arabidopsis physiology, Genotype, Hydrogen-Ion Concentration, Light, Photosynthesis genetics, Photosynthesis radiation effects, Photosystem I Protein Complex radiation effects, Photosystem II Protein Complex radiation effects, Arabidopsis Proteins genetics, Membrane Proteins genetics, Photosynthetic Reaction Center Complex Proteins genetics, Photosystem I Protein Complex genetics, Photosystem II Protein Complex genetics
- Abstract
Although light is essential for photosynthesis, when in excess, it may damage the photosynthetic apparatus, leading to a phenomenon known as photoinhibition. Photoinhibition was thought as a light-induced damage to photosystem II; however, it is now clear that even photosystem I may become very vulnerable to light. One main characteristic of light induced damage to photosystem II (PSII) is the increased turnover of the reaction center protein, D1: when rate of degradation exceeds the rate of synthesis, loss of PSII activity is observed. With respect to photosystem I (PSI), an excess of electrons, instead of an excess of light, may be very dangerous. Plants possess a number of mechanisms able to prevent, or limit, such damages by safe thermal dissipation of light energy (non-photochemical quenching, NPQ), slowing-down of electron transfer through the intersystem transport chain (photosynthesis-control, PSC) in co-operation with the Proton Gradient Regulation (PGR) proteins, PGR5 and PGRL1, collectively called as short-term photoprotection mechanisms, and the redistribution of light between photosystems, called state transitions (responsible of fluorescence quenching at PSII, qT), is superimposed to these short term photoprotective mechanisms. In this manuscript we have generated a number of higher order mutants by crossing genotypes carrying defects in each of the short-term photoprotection mechanisms, with the final aim to obtain a direct comparison of their role and efficiency in photoprotection. We found that mutants carrying a defect in the ΔpH-dependent photosynthesis-control are characterized by photoinhibition of both photosystems, irrespectively of whether PSBS-dependent NPQ or state transitions defects were present or not in the same individual, demonstrating the primary role of PSC in photoprotection. Moreover, mutants with a limited capability to develop a strong PSBS-dependent NPQ, were characterized by a high turnover of the D1 protein and high values of Y(NO), which might reflect energy quenching processes occurring within the PSII reaction center.
- Published
- 2020
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33. A New Remote Sensing-Based System for the Monitoring and Analysis of Growth and Gas Exchange Rates of Photosynthetic Microorganisms Under Simulated Non-Terrestrial Conditions.
- Author
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Battistuzzi M, Cocola L, Salasnich B, Erculiani MS, Alei E, Morosinotto T, Claudi R, Poletto L, and La Rocca N
- Abstract
Oxygenic photosynthetic microorganisms are a focal point of research in the context of human space exploration. As part of the bioregenerative life-support systems, they could have a key role in the production of breathable O
2 , edible biomasses and in the regeneration of CO2 rich-atmospheres and wastewaters produced by astronauts. The test of the organism's response to simulated physico-chemical parameters of planetary bodies could also provide important information about their habitability potential. It is believed that the success of future planetary and space missions will require innovative technologies, developed on the base of preliminary experiments in custom-made laboratory facilities. In this context, simulation chambers will play a pivotal role by allowing the growth of the microorganisms under controlled conditions and the evaluation in real-time of their biomass productivity and impact on atmosphere composition. We here present a system capable of addressing these requirements with high replicability and low costs. The setup is composed by three main parts: 1) a Star Light Simulator, able to generate different light intensities and spectra, including those of non-solar stars; 2) an Atmosphere Simulator Chamber where cultures of photosynthetic microorganisms can be exposed to different gas compositions; 3) a reflectivity detection system to measure from remote the Normalized Difference Vegetation Indexes (NDVI). Such a setup allows us to monitor photosynthetic microorganism's growth and gas exchange performances under selected conditions of light quality and intensity, temperature, pressure, and atmospheres simulating non-terrestrial environments. All parameters are detected by remote sensing techniques, thus without interfering with the experiments and altering the environmental conditions set. We validated the setup by growing cyanobacteria liquid cultures under different light intensities of solar illumination, collecting data on their growth rate, photosynthetic activity, and gas exchange capacity. We utilized the reflectivity detection system to measure the reflection spectra of the growing cultures, obtaining their relative NDVI that was shown to correlate with optical density, chlorophyll content, and dry weight, demonstrating the potential application of this index as a proxy of growth., (Copyright © 2020 Battistuzzi, Cocola, Salasnich, Erculiani, Alei, Morosinotto, Claudi, Poletto and La Rocca.)- Published
- 2020
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34. Photosynthesis Regulation in Response to Fluctuating Light in the Secondary Endosymbiont Alga Nannochloropsis gaditana.
- Author
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Bellan A, Bucci F, Perin G, Alboresi A, and Morosinotto T
- Subjects
- Biodiversity, Electron Transport physiology, Oxidative Stress, Photosystem I Protein Complex metabolism, Photosystem I Protein Complex radiation effects, Plants metabolism, Stramenopiles growth & development, Stramenopiles radiation effects, Light, Photosynthesis physiology, Stramenopiles metabolism, Symbiosis physiology
- Abstract
In nature, photosynthetic organisms are exposed to highly dynamic environmental conditions where the excitation energy and electron flow in the photosynthetic apparatus need to be continuously modulated. Fluctuations in incident light are particularly challenging because they drive oversaturation of photosynthesis with consequent oxidative stress and photoinhibition. Plants and algae have evolved several mechanisms to modulate their photosynthetic machinery to cope with light dynamics, such as thermal dissipation of excited chlorophyll states (non-photochemical quenching, NPQ) and regulation of electron transport. The regulatory mechanisms involved in the response to light dynamics have adapted during evolution, and exploring biodiversity is a valuable strategy for expanding our understanding of their biological roles. In this work, we investigated the response to fluctuating light in Nannochloropsis gaditana, a eukaryotic microalga of the phylum Heterokonta originating from a secondary endosymbiotic event. Nannochloropsis gaditana is negatively affected by light fluctuations, leading to large reductions in growth and photosynthetic electron transport. Exposure to light fluctuations specifically damages photosystem I, likely because of the ineffective regulation of electron transport in this species. The role of NPQ, also assessed using a mutant strain specifically depleted of this response, was instead found to be minor, especially in responding to the fastest light fluctuations., (� The Author(s) 2019. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
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35. Role and regulation of class-C flavodiiron proteins in photosynthetic organisms.
- Author
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Alboresi A, Storti M, Cendron L, and Morosinotto T
- Subjects
- Bacterial Proteins genetics, Cyanobacteria genetics, Oxidation-Reduction, Photosystem I Protein Complex genetics, Bacterial Proteins metabolism, Cyanobacteria metabolism, Nitrogen Fixation physiology, Oxygen metabolism, Photosynthesis physiology, Photosystem I Protein Complex metabolism
- Abstract
The regulation of photosynthesis is crucial to efficiently support the assimilation of carbon dioxide and to prevent photodamage. One key regulatory mechanism is the pseudo-cyclic electron flow (PCEF) mediated by class-C flavodiiron proteins (FLVs). These enzymes use electrons coming from Photosystem I (PSI) to reduce oxygen to water, preventing over-reduction in the acceptor side of PSI. FLVs are widely distributed among organisms performing oxygenic photosynthesis and they have been shown to be fundamental in many different conditions such as fluctuating light, sulfur deprivation and plant submersion. Moreover, since FLVs reduce oxygen they can help controlling the redox status of the cell and maintaining the microoxic environment essential for processes such as nitrogen fixation in cyanobacteria. Despite these important roles identified in various species, the genes encoding for FLV proteins have been lost in angiosperms where their activity could have been at least partially compensated by a more efficient cyclic electron flow (CEF). The present work reviews the information emerged on FLV function, analyzing recent structural data that suggest FLV could be regulated through a conformational change., (© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)
- Published
- 2019
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36. Thylakoid Protein Phosphorylation Dynamics in a Moss Mutant Lacking SERINE/THREONINE PROTEIN KINASE STN8.
- Author
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Gerotto C, Trotta A, Bajwa AA, Mancini I, Morosinotto T, and Aro EM
- Subjects
- Arabidopsis genetics, Arabidopsis metabolism, Bryopsida genetics, Chloroplasts genetics, Chloroplasts ultrastructure, Kinetics, Light, Light-Harvesting Protein Complexes genetics, Light-Harvesting Protein Complexes metabolism, Microscopy, Electron, Transmission, Mutation, Phosphorylation, Photosynthesis genetics, Photosynthesis radiation effects, Photosystem II Protein Complex genetics, Photosystem II Protein Complex metabolism, Plant Proteins genetics, Protein Kinases genetics, Protein Kinases metabolism, Protein Serine-Threonine Kinases genetics, Thylakoids genetics, Thylakoids ultrastructure, Bryopsida metabolism, Chloroplasts metabolism, Plant Proteins metabolism, Protein Serine-Threonine Kinases metabolism, Thylakoids metabolism
- Abstract
In all eukaryotes, protein phosphorylation is a key regulatory mechanism in several cellular processes, including the acclimation of photosynthesis to environmental cues. Despite being a well-conserved regulatory mechanism in the chloroplasts of land plants, distinct differences in thylakoid protein phosphorylation patterns have emerged from studies on species of different phylogenetic groups. Here, we analyzed thylakoid protein phosphorylation in the moss Physcomitrella patens , assessing the thylakoid phospho-protein profile and dynamics in response to changes in white light intensity. Compared with Arabidopsis ( Arabidopsis thaliana ), parallel characterization of wild-type P patens and the knockout mutant stn8 (depleted in SER/THR PROTEIN KINASE8 [STN8]) disclosed a moss-specific pattern of thylakoid protein phosphorylation, both with respect to specific targets and to their dynamic phosphorylation in response to environmental cues. Unlike vascular plants, (1) phosphorylation of the PSII protein D1 in P patens was negligible under all light conditions, (2) phosphorylation of the PSII core subunits CP43 and D2 showed only minor changes upon fluctuations in light intensity, and (3) absence of STN8 completely abolished all PSII core protein phosphorylation. Further, we detected light-induced phosphorylation in the minor light harvesting complex (LHC) antenna protein LHCB6, which was dependent on STN8 kinase activity, and found specific phosphorylations on LHCB3. Data presented here provide further insights into the appearance and physiological role of thylakoid protein phosphorylation during evolution of oxygenic photosynthetic organisms and their colonization of land., (© 2019 American Society of Plant Biologists. All Rights Reserved.)
- Published
- 2019
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37. Role of cyclic and pseudo-cyclic electron transport in response to dynamic light changes in Physcomitrella patens.
- Author
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Storti M, Alboresi A, Gerotto C, Aro EM, Finazzi G, and Morosinotto T
- Subjects
- Bryopsida genetics, Chloroplasts metabolism, Electron Transport genetics, Light, Membrane Proteins genetics, Membrane Proteins metabolism, Mutation, Photosynthesis genetics, Photosynthesis physiology, Photosynthetic Reaction Center Complex Proteins metabolism, Photosystem I Protein Complex genetics, Plants, Genetically Modified, Sunlight, Thylakoids metabolism, Bryopsida physiology, Electron Transport physiology, Photosynthetic Reaction Center Complex Proteins genetics, Photosystem I Protein Complex metabolism
- Abstract
Photosynthetic organisms support cell metabolism by harvesting sunlight and driving the electron transport chain at the level of thylakoid membranes. Excitation energy and electron flow in the photosynthetic apparatus is continuously modulated in response to dynamic environmental conditions. Alternative electron flow around photosystem I plays a seminal role in this regulation contributing to photoprotection by mitigating overreduction of the electron carriers. Different pathways of alternative electron flow coexist in the moss Physcomitrella patens, including cyclic electron flow mediated by the PGRL1/PGR5 complex and pseudo-cyclic electron flow mediated by the flavodiiron proteins FLV. In this work, we generated P. patens plants carrying both pgrl1 and flva knock-out mutations. A comparative analysis of the WT, pgrl1, flva, and pgrl1 flva lines suggests that cyclic and pseudo-cyclic processes have a synergic role in the regulation of photosynthetic electron transport. However, although both contribute to photosystem I protection from overreduction by modulating electron flow following changes in environmental conditions, FLV activity is particularly relevant in the first seconds after a light change whereas PGRL1 has a major role upon sustained strong illumination., (© 2018 John Wiley & Sons Ltd.)
- Published
- 2019
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38. The potential of quantitative models to improve microalgae photosynthetic efficiency.
- Author
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Perin G, Bellan A, Bernardi A, Bezzo F, and Morosinotto T
- Subjects
- Biomass, Biotechnology, Carbon Dioxide metabolism, Microalgae metabolism, Photosynthesis physiology
- Abstract
The massive increase in carbon dioxide concentration in the atmosphere driven by human activities is causing huge negative consequences and new sustainable sources of energy, food and materials are highly needed. Algae are unicellular photosynthetic microorganisms that can provide a highly strategic contribution to this challenge as alternative source of biomass to complement crops cultivation. Algae industrial cultures are commonly limited by light availability, and biomass accumulation is strongly dependent on their photon-to-biomass conversion efficiency. Investigation of algae photosynthetic metabolism is thus strategic for the generation of more efficient strains with higher productivity. Algae are cultivated at industrial scale in conditions highly different from the natural niches they adapted to and strains development efforts must fully consider the seminal influence on productivity of regulatory mechanism of photosynthesis as well as of cultivation parameters like cells concentration, light distribution in the culture, mixing, nutrients and carbon dioxide availability. In this review we will focus in particular on how mathematical models can account for the complex influence of all environmental parameters and can be exploited for development of improved algae strains., (© 2018 Scandinavian Plant Physiology Society.)
- Published
- 2019
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39. Plant biodiversity and regulation of photosynthesis in the natural environment.
- Author
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Sello S, Meneghesso A, Alboresi A, Baldan B, and Morosinotto T
- Subjects
- Electron Transport, Environment, Light, Photosystem II Protein Complex metabolism, Biodiversity, Photosynthesis physiology, Plants metabolism
- Abstract
Main Conclusion: Investigation of photosynthesis regulation in different plant groups exposed to variable conditions showed that all species have similar photosynthetic electron transport modulation while excess energy dissipation is species specific. Photosynthesis is regulated in response to dynamic environmental conditions to satisfy plant metabolic demands while also avoiding possible over-excitation of the electron transport chain and the generation of harmful reactive oxygen species. Photosynthetic organisms evolved several mechanisms to modulate light harvesting and electron transport efficiency to respond to conditions changing at different timescales, going from fast sun flecks to slow seasonal variations. These regulatory mechanisms changed during evolution of photosynthetic organisms, also adapting to various ecological niches, making the investigation of plant biodiversity highly valuable to uncover conserved traits and plasticity of photosynthetic regulation and complement studies on model species. In this work, a set of plants belonging to different genera of angiosperms, gymnosperms, ferns and lycophytes were investigated by monitoring their photosynthetic parameters in different seasons looking for common trends and differences. In all plants, analysed photosynthetic electron transport rate was found to be modulated by growth light intensity, ensuring a balance between available energy and photochemical capacity. Growth light also influenced the threshold where heat dissipation of excitation energy, a mechanism called non-photochemical quenching (NPQ), was activated. On the contrary, NPQ amplitude did not correlate with light intensity experienced by the plants but was a species-specific feature. The zeaxanthin-dependent component of NPQ, qZ, was found to be the most variable in different plants and its modulation influenced the intensity and the kinetic properties of the response.
- Published
- 2019
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40. Balancing protection and efficiency in the regulation of photosynthetic electron transport across plant evolution.
- Author
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Alboresi A, Storti M, and Morosinotto T
- Subjects
- Magnoliopsida physiology, Photosystem I Protein Complex metabolism, Biological Evolution, Electron Transport, Photosynthesis physiology, Plant Physiological Phenomena, Plant Proteins metabolism
- Abstract
Contents Summary 105 I. Introduction 105 II. Diversity of molecular mechanisms for regulation of photosynthetic electron transport 106 III. Role of FLVs in the regulation of photosynthesis in eukaryotes 107 IV. Why were FLVs lost in angiosperms? 108 V. Conclusions 108 Acknowledgements 109 References 109 SUMMARY: Photosynthetic electron transport requires continuous modulation to maintain the balance between light availability and metabolic demands. Multiple mechanisms for the regulation of electron transport have been identified and are unevenly distributed among photosynthetic organisms. Flavodiiron proteins (FLVs) influence photosynthetic electron transport by accepting electrons downstream of photosystem I to reduce oxygen to water. FLV activity has been demonstrated in cyanobacteria, green algae and mosses to be important in avoiding photosystem I overreduction upon changes in light intensity. FLV-encoding sequences were nevertheless lost during evolution by angiosperms, suggesting that these plants increased the efficiency of other mechanisms capable of accepting electrons from photosystem I, making the FLV activity for protection from overreduction superfluous or even detrimental for photosynthetic efficiency., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)
- Published
- 2019
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41. Global spectroscopic analysis to study the regulation of the photosynthetic proton motive force: A critical reappraisal.
- Author
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Allorent G, Byrdin M, Carraretto L, Morosinotto T, Szabo I, and Finazzi G
- Subjects
- Arabidopsis genetics, Arabidopsis radiation effects, Light, Light-Harvesting Protein Complexes genetics, Photosynthesis, Plants, Genetically Modified genetics, Plants, Genetically Modified radiation effects, Potassium Channels genetics, Thylakoids metabolism, Arabidopsis metabolism, Chlorophyll metabolism, Light-Harvesting Protein Complexes metabolism, Plants, Genetically Modified metabolism, Potassium Channels metabolism, Proton-Motive Force
- Abstract
In natural variable environments, plants rapidly adjust photosynthesis for optimum balance between photochemistry and photoprotection. These adjustments mainly occur via changes in their proton motive force (pmf). Recent studies based on time resolved analysis of the Electro Chromic Signal (ECS) bandshift of photosynthetic pigments in the model plant Arabidopsis thaliana have suggested an active role of ion fluxes across the thylakoid membranes in the regulation of the pmf. Among the different channels and transporters possibly involved in this phenomenon, we previously identified the TPK3 potassium channel. Plants silenced for TPK3 expression displayed light stress signatures, with reduced Non Photochemical Quenching (NPQ) capacity and sustained anthocyanin accumulation, even at moderate intensities. In this work we re-examined the role of this protein in pmf regulation, starting from the observation that both TPK3 knock-down (TPK3 KD) and WT plants display enhanced anthocyanin accumulation in the light under certain growth conditions, especially in old leaves. We thus compared the pmf features of young "green" (without anthocyanins) and old "red" (with anthocyanins) leaves in both genotypes using a global fit analysis of the ECS. We found that the differences in the ECS profile measured between the two genotypes reflect not only differences in TPK3 expression level, but also a modified photosynthetic activity of stressed red leaves, which are present in a larger amounts in the TPK3 KD plants., (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Published
- 2018
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42. Systemic Calcium Wave Propagation in Physcomitrella patens.
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Storti M, Costa A, Golin S, Zottini M, Morosinotto T, and Alboresi A
- Subjects
- Arabidopsis metabolism, Bryopsida cytology, Bryopsida physiology, Calcium analysis, Calcium metabolism, Calmodulin metabolism, Dehydration, Fluorescence Resonance Energy Transfer, Luminescent Proteins metabolism, Molecular Imaging methods, Osmotic Pressure, Plant Cells metabolism, Plants, Genetically Modified, Recombinant Fusion Proteins metabolism, Bryopsida metabolism, Calcium Signaling
- Abstract
The adaptation to dehydration and rehydration cycles represents a key step in the evolution of photosynthetic organisms and requires the development of mechanisms by which to sense external stimuli and translate them into signaling components. In this study, we used genetically encoded fluorescent sensors to detect specific transient increases in the Ca2+ concentration in the moss Physcomitrella patens upon dehydration and rehydration treatment. Observation of the entire plant in a single time-series acquisition revealed that various cell types exhibited different sensitivities to osmotic stress and that Ca2+ waves originated from the basal part of the gametophore and were directionally propagated towards the top of the plant. Under similar conditions, the vascular plant Arabidopsis thaliana exhibited Ca2+ waves that propagated at a higher speed than those of P. patens. Our results suggest that systemic Ca2+ propagation occurs in plants even in the absence of vascular tissue, even though the rates can be different.
- Published
- 2018
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43. Transcriptome and Cell Physiological Analyses in Different Rice Cultivars Provide New Insights Into Adaptive and Salinity Stress Responses.
- Author
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Formentin E, Sudiro C, Perin G, Riccadonna S, Barizza E, Baldoni E, Lavezzo E, Stevanato P, Sacchi GA, Fontana P, Toppo S, Morosinotto T, Zottini M, and Lo Schiavo F
- Abstract
Salinity tolerance has been extensively investigated in recent years due to its agricultural importance. Several features, such as the regulation of ionic transporters and metabolic adjustments, have been identified as salt tolerance hallmarks. Nevertheless, due to the complexity of the trait, the results achieved to date have met with limited success in improving the salt tolerance of rice plants when tested in the field, thus suggesting that a better understanding of the tolerance mechanisms is still required. In this work, differences between two varieties of rice with contrasting salt sensitivities were revealed by the imaging of photosynthetic parameters, ion content analysis and a transcriptomic approach. The transcriptomic analysis conducted on tolerant plants supported the setting up of an adaptive program consisting of sodium distribution preferentially limited to the roots and older leaves, and in the activation of regulatory mechanisms of photosynthesis in the new leaves. As a result, plants resumed grow even under prolonged saline stress. In contrast, in the sensitive variety, RNA-seq analysis revealed a misleading response, ending in senescence and cell death. The physiological response at the cellular level was investigated by measuring the intracellular profile of H
2 O2 in the roots, using a fluorescent probe. In the roots of tolerant plants, a quick response was observed with an increase in H2 O2 production within 5 min after salt treatment. The expression analysis of some of the genes involved in perception, signal transduction and salt stress response confirmed their early induction in the roots of tolerant plants compared to sensitive ones. By inhibiting the synthesis of apoplastic H2 O2 , a reduction in the expression of these genes was detected. Our results indicate that quick H2 O2 signaling in the roots is part of a coordinated response that leads to adaptation instead of senescence in salt-treated rice plants.- Published
- 2018
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44. Mitochondria Affect Photosynthetic Electron Transport and Photosensitivity in a Green Alga.
- Author
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Larosa V, Meneghesso A, La Rocca N, Steinbeck J, Hippler M, Szabò I, and Morosinotto T
- Subjects
- Chloroplasts genetics, Chloroplasts metabolism, Electron Transport genetics, Light, Mutation, NADH Dehydrogenase genetics, NADH Dehydrogenase metabolism, Photosystem I Protein Complex metabolism, Photosystem II Protein Complex metabolism, Plastoquinone metabolism, Chlamydomonas reinhardtii physiology, Mitochondria metabolism, Photosynthesis physiology
- Abstract
Photosynthetic organisms use sunlight as the primary source of energy to support their metabolism. In eukaryotes, reactions responsible of the conversion of light into chemical energy occur in specific organelles, the chloroplasts. In this study, we showed that mitochondria also have a seminal influence on cells' energy metabolism and on photosynthetic reactions. This is illustrated by the observation that the strong photosensitivity of Chlamydomonas reinhardtii cells depleted of the chloroplast protein PGRL1 was rescued by the introduction of a mitochondrial mutation affecting respiratory complex I. Functional analysis showed that such a reduced respiratory activity influenced chloroplast electron transport with consequent overreduction of plastoquinone and donor-side limitation of photosystem I (PSI). As a consequence, damage due to excess light affected more photosystem II (PSII) rather than PSI. Double mutant cells are able to grow under excess illumination, while single pgrl1 are not, thanks to the presence of an efficient repair mechanism of PSII. These results also underline the seminal biological relevance of the regulation of electron transport reactions within the photosynthetic complexes. Photosynthetic organisms evolved a strategy to respond to excess light where damage is targeting preferentially to a specific complex, PSII. Cells are able to endure extensive damage targeting this complex thanks to an efficient repair mechanisms, while if PSI is affected, there are drastic consequences on growth., (© 2018 American Society of Plant Biologists. All Rights Reserved.)
- Published
- 2018
- Full Text
- View/download PDF
45. A mathematical model to guide genetic engineering of photosynthetic metabolism.
- Author
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Perin G, Bernardi A, Bellan A, Bezzo F, and Morosinotto T
- Subjects
- Genetic Engineering, Models, Biological, Photosynthesis physiology, Stramenopiles genetics, Stramenopiles metabolism
- Abstract
The optimization of algae biomass productivity in industrial cultivation systems requires genetic improvement of wild type strains isolated from nature. One of the main factors affecting algae productivity is their efficiency in converting light into chemical energy and this has been a major target of recent genetic efforts. However, photosynthetic productivity in algae cultures depends on many environmental parameters, making the identification of advantageous genotypes complex and the achievement of concrete improvements slow. In this work, we developed a mathematical model to describe the key factors influencing algae photosynthetic productivity in a photobioreactor, using experimental measurements for the WT strain of Nannochloropsis gaditana. The model was then exploited to predict the effect of potential genetic modifications on algae performances in an industrial context, showing the ability to predict the productivity of mutants with specific photosynthetic phenotypes. These results show that a quantitative model can be exploited to identify the genetic modifications with the highest impact on productivity taking into full account the complex influence of environmental conditions, efficiently guiding engineering efforts., (Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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46. Semi-empirical modeling of microalgae photosynthesis in different acclimation states - Application to N. gaditana.
- Author
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Bernardi A, Nikolaou A, Meneghesso A, Chachuat B, Morosinotto T, and Bezzo F
- Subjects
- Acclimatization, Microalgae metabolism, Microalgae physiology, Models, Biological, Photosynthesis physiology, Stramenopiles metabolism, Stramenopiles physiology
- Abstract
The development of mathematical models capable of accurate predictions of the photosynthetic productivity of microalgae under variable light conditions is paramount to the development of large-scale production systems. The process of photoacclimation is particularly important in outdoor cultivation systems, whereby seasonal variation of the light irradiance can greatly influence microalgae growth. This paper presents a dynamic model that captures the effect of photoacclimation on the photosynthetic production. It builds upon an existing semi-empirical model describing the processes of photoproduction, photoregulation and photoinhibition via the introduction of acclimation rules for key parameters. The model is calibrated against a dataset comprising pulsed amplitude modulation fluorescence, photosynthesis rate, and antenna size measurements for the microalga Nannochloropsis gaditana in several acclimation states. It is shown that the calibrated model is capable of accurate predictions of fluorescence and respirometry data, both in interpolation and in extrapolation., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2017
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47. Photoprotection strategies of the alga Nannochloropsis gaditana.
- Author
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Chukhutsina VU, Fristedt R, Morosinotto T, and Croce R
- Subjects
- Algal Proteins chemistry, Algal Proteins isolation & purification, Algal Proteins physiology, Fluorescence, Light, Light-Harvesting Protein Complexes chemistry, Light-Harvesting Protein Complexes physiology, Photosystem I Protein Complex metabolism, Photosystem II Protein Complex metabolism, Radiation Tolerance physiology, Spectrometry, Fluorescence, Stramenopiles chemistry, Stramenopiles radiation effects, Xanthophylls chemistry, Stramenopiles physiology
- Abstract
Nannochloropsis spp. are algae with high potential for biotechnological applications due to their capacity to accumulate lipids. However, little is known about their photosynthetic apparatus and acclimation/photoprotective strategies. In this work, we studied the mechanisms of non-photochemical quenching (NPQ), the fast response to high light stress, in Nannochloropsis gaditana by "locking" the cells in six different states during quenching activation and relaxation. Combining biochemical analysis with time-resolved fluorescence spectroscopy, we correlated each NPQ state with the presence of two well-known NPQ components: de-epoxidized xanthophylls and stress-related antenna proteins (LHCXs). We demonstrated that after exposure to strong light, the rapid quenching that takes place in the antennas of both photosystems was associated with the presence of LHCXs. At later stages, quenching occurs mainly in the antennas of PSII and correlates with the amount of de-epoxidised xanthophylls. We also observed changes in the distribution of excitation energy between photosystems, which suggests redistribution of excitation between photosystems as part of the photo-protective strategy. A multistep model for NPQ induction and relaxation in N. gaditana is discussed., (Copyright © 2017. Published by Elsevier B.V.)
- Published
- 2017
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48. Alternative electron transport mediated by flavodiiron proteins is operational in organisms from cyanobacteria up to gymnosperms.
- Author
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Ilík P, Pavlovič A, Kouřil R, Alboresi A, Morosinotto T, Allahverdiyeva Y, Aro EM, Yamamoto H, and Shikanai T
- Subjects
- Cyanobacteria radiation effects, Cycadopsida radiation effects, Electron Transport, Kinetics, Light, Oxidation-Reduction, Photosynthesis radiation effects, Phylogeny, Cyanobacteria metabolism, Cycadopsida metabolism, Flavoproteins metabolism
- Abstract
Photo-reduction of O
2 to water mediated by flavodiiron proteins (FDPs) represents a safety valve for the photosynthetic electron transport chain in fluctuating light. So far, the FDP-mediated O2 photo-reduction has been evidenced only in cyanobacteria and the moss Physcomitrella; however, a recent phylogenetic analysis of transcriptomes of photosynthetic organisms has also revealed the presence of FDP genes in several nonflowering plant groups. What remains to be clarified is whether the FDP-dependent O2 photo-reduction is actually operational in these organisms. We have established a simple method for the monitoring of FDP-mediated O2 photo-reduction, based on the measurement of redox kinetics of P700 (the electron donor of photosystem I) upon dark-to-light transition. The O2 photo-reduction is manifested as a fast re-oxidation of P700. The validity of the method was verified by experiments with transgenic organisms, namely FDP knock-out mutants of Synechocystis and Physcomitrella and transgenic Arabidopsis plants expressing FDPs from Physcomitrella. We observed the fast P700 re-oxidation in representatives of all green plant groups excluding angiosperms. Our results provide strong evidence that the FDP-mediated O2 photo-reduction is functional in all nonflowering green plant groups. This finding suggests a major change in the strategy of photosynthetic regulation during the evolution of angiosperms., (© 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.)- Published
- 2017
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49. A Palmitic Acid Elongase Affects Eicosapentaenoic Acid and Plastidial Monogalactosyldiacylglycerol Levels in Nannochloropsis.
- Author
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Dolch LJ, Rak C, Perin G, Tourcier G, Broughton R, Leterrier M, Morosinotto T, Tellier F, Faure JD, Falconet D, Jouhet J, Sayanova O, Beaudoin F, and Maréchal E
- Subjects
- Acetyltransferases genetics, Algal Proteins genetics, Cloning, Molecular, Eicosapentaenoic Acid genetics, Fatty Acids, Unsaturated metabolism, Fluorescence, Gene Expression Regulation, Plant, Photosynthesis, Phylogeny, Plant Proteins genetics, Plants, Genetically Modified, Sphingolipids metabolism, Stramenopiles genetics, Thylakoids genetics, Thylakoids ultrastructure, Triglycerides metabolism, Yeasts genetics, Acetyltransferases metabolism, Algal Proteins metabolism, Eicosapentaenoic Acid metabolism, Galactolipids metabolism, Plant Proteins metabolism, Plastids metabolism, Stramenopiles metabolism
- Abstract
Nannochloropsis species are oleaginous eukaryotes containing a plastid limited by four membranes, deriving from a secondary endosymbiosis. In Nannochloropsis, thylakoid lipids, including monogalactosyldiacylglycerol (MGDG), are enriched in eicosapentaenoic acid (EPA). The need for EPA in MGDG is not understood. Fatty acids are de novo synthesized in the stroma, then converted into very-long-chain polyunsaturated fatty acids (FAs) at the endoplasmic reticulum (ER). The production of MGDG relies therefore on an EPA supply from the ER to the plastid, following an unknown process. We identified seven elongases and five desaturases possibly involved in EPA production in Nannochloropsis gaditana Among the six heterokont-specific saturated FA elongases possibly acting upstream in this pathway, we characterized the highly expressed isoform Δ0-ELO1 Heterologous expression in yeast (Saccharomyces cerevisiae) showed that NgΔ0-ELO1 could elongate palmitic acid. Nannochloropsis Δ0-elo1 mutants exhibited a reduced EPA level and a specific decrease in MGDG In NgΔ0-elo1 lines, the impairment of photosynthesis is consistent with a role of EPA-rich MGDG in nonphotochemical quenching control, possibly providing an appropriate MGDG platform for the xanthophyll cycle. Concomitantly with MGDG decrease, the level of triacylglycerol (TAG) containing medium chain FAs increased. In Nannochloropsis, part of EPA used for MGDG production is therefore biosynthesized by a channeled process initiated at the elongation step of palmitic acid by Δ0-ELO1, thus acting as a committing enzyme for galactolipid production. Based on the MGDG/TAG balance controlled by Δ0-ELO1, this study also provides novel prospects for the engineering of oleaginous microalgae for biotechnological applications., (© 2017 American Society of Plant Biologists. All Rights Reserved.)
- Published
- 2017
- Full Text
- View/download PDF
50. Conservation of core complex subunits shaped the structure and function of photosystem I in the secondary endosymbiont alga Nannochloropsis gaditana.
- Author
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Alboresi A, Le Quiniou C, Yadav SK, Scholz M, Meneghesso A, Gerotto C, Simionato D, Hippler M, Boekema EJ, Croce R, and Morosinotto T
- Subjects
- Amino Acid Sequence, Light-Harvesting Protein Complexes chemistry, Light-Harvesting Protein Complexes metabolism, Light-Harvesting Protein Complexes ultrastructure, Models, Biological, Photosystem I Protein Complex ultrastructure, Pigments, Biological metabolism, Protein Subunits chemistry, Spectrometry, Fluorescence, Thylakoids metabolism, Conserved Sequence, Photosystem I Protein Complex chemistry, Photosystem I Protein Complex metabolism, Protein Subunits metabolism, Stramenopiles metabolism, Symbiosis
- Abstract
Photosystem I (PSI) is a pigment protein complex catalyzing the light-driven electron transport from plastocyanin to ferredoxin in oxygenic photosynthetic organisms. Several PSI subunits are highly conserved in cyanobacteria, algae and plants, whereas others are distributed differentially in the various organisms. Here we characterized the structural and functional properties of PSI purified from the heterokont alga Nannochloropsis gaditana, showing that it is organized as a supercomplex including a core complex and an outer antenna, as in plants and other eukaryotic algae. Differently from all known organisms, the N. gaditana PSI supercomplex contains five peripheral antenna proteins, identified by proteome analysis as type-R light-harvesting complexes (LHCr4-8). Two antenna subunits are bound in a conserved position, as in PSI in plants, whereas three additional antennae are associated with the core on the other side. This peculiar antenna association correlates with the presence of PsaF/J and the absence of PsaH, G and K in the N. gaditana genome and proteome. Excitation energy transfer in the supercomplex is highly efficient, leading to a very high trapping efficiency as observed in all other PSI eukaryotes, showing that although the supramolecular organization of PSI changed during evolution, fundamental functional properties such as trapping efficiency were maintained., (© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.)
- Published
- 2017
- Full Text
- View/download PDF
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