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      Lhcx proteins provide photoprotection via thermal dissipation of absorbed light in the diatom Phaeodactylum tricornutum

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          Abstract

          Diatoms possess an impressive capacity for rapidly inducible thermal dissipation of excess absorbed energy (qE), provided by the xanthophyll diatoxanthin and Lhcx proteins. By knocking out the Lhcx1 and Lhcx2 genes individually in Phaeodactylum tricornutum strain 4 and complementing the knockout lines with different Lhcx proteins, multiple mutants with varying qE capacities are obtained, ranging from zero to high values. We demonstrate that qE is entirely dependent on the concerted action of diatoxanthin and Lhcx proteins, with Lhcx1, Lhcx2 and Lhcx3 having similar functions. Moreover, we establish a clear link between Lhcx1/2/3 mediated inducible thermal energy dissipation and a reduction in the functional absorption cross-section of photosystem II. This regulation of the functional absorption cross-section can be tuned by altered Lhcx protein expression in response to environmental conditions. Our results provide a holistic understanding of the rapidly inducible thermal energy dissipation process and its mechanistic implications in diatoms.

          Abstract

          Photosynthetic organisms can dissipate excess absorbed light energy as heat to avoid photodamage. Here the authors show that induced thermal dissipation in the diatom Phaeodactylum tricornutum Pt4 is Lhcx protein-dependent and correlates with a reduced functional absorption cross-section of photosystem II.

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          New fluorescence parameters for the determination of q(a) redox state and excitation energy fluxes.

          A number of useful photosynthetic parameters are commonly derived from saturation pulse-induced fluorescence analysis. We show, that q(P), an estimate of the fraction of open centers, is based on a pure 'puddle' antenna model, where each Photosystem (PS) II center possesses its own independent antenna system. This parameter is incompatible with more realistic models of the photosynthetic unit, where reaction centers are connected by shared antenna, that is, the so-called 'lake' or 'connected units' models. We thus introduce a new parameter, q(L), based on a Stern-Volmer approach using a lake model, which estimates the fraction of open PS II centers. We suggest that q(L) should be a useful parameter for terrestrial plants consistent with a high connectivity of PS II units, whereas some marine species with distinct antenna architecture, may require the use of more complex parameters based on intermediate models of the photosynthetic unit. Another useful parameter calculated from fluorescence analysis is Phi(II), the yield of PS II. In contrast to q(L), we show that the Phi(II) parameter can be derived from either a pure 'lake' or pure 'puddle' model, and is thus likely to be a robust parameter. The energy absorbed by PS II is divided between the fraction used in photochemistry, Phi(II), and that lost non-photochemically. We introduce two additional parameters that can be used to estimate the flux of excitation energy into competing non-photochemical pathways, the yield induced by downregulatory processes, Phi(NPQ), and the yield for other energy losses, Phi(NO).
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            Comprehensive algorithm for quantitative real-time polymerase chain reaction.

            Quantitative real-time polymerase chain reactions (qRT-PCR) have become the method of choice for rapid, sensitive, quantitative comparison of RNA transcript abundance. Useful data from this method depend on fitting data to theoretical curves that allow computation of mRNA levels. Calculating accurate mRNA levels requires important parameters such as reaction efficiency and the fractional cycle number at threshold (CT) to be used; however, many algorithms currently in use estimate these important parameters. Here we describe an objective method for quantifying qRT-PCR results using calculations based on the kinetics of individual PCR reactions without the need of the standard curve, independent of any assumptions or subjective judgments which allow direct calculation of efficiency and CT. We use a four-parameter logistic model to fit the raw fluorescence data as a function of PCR cycles to identify the exponential phase of the reaction. Next, we use a three-parameter simple exponent model to fit the exponential phase using an iterative nonlinear regression algorithm. Within the exponential portion of the curve, our technique automatically identifies candidate regression values using the P-value of regression and then uses a weighted average to compute a final efficiency for quantification. For CT determination, we chose the first positive second derivative maximum from the logistic model. This algorithm provides an objective and noise-resistant method for quantification of qRT-PCR results that is independent of the specific equipment used to perform PCR reactions.
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              A pigment-binding protein essential for regulation of photosynthetic light harvesting.

              Photosynthetic light harvesting in plants is regulated in response to changes in incident light intensity. Absorption of light that exceeds a plant's capacity for fixation of CO2 results in thermal dissipation of excitation energy in the pigment antenna of photosystem II by a poorly understood mechanism. This regulatory process, termed nonphotochemical quenching, maintains the balance between dissipation and utilization of light energy to minimize generation of oxidizing molecules, thereby protecting the plant against photo-oxidative damage. To identify specific proteins that are involved in nonphotochemical quenching, we have isolated mutants of Arabidopsis thaliana that cannot dissipate excess absorbed light energy. Here we show that the gene encoding PsbS, an intrinsic chlorophyll-binding protein of photosystem II, is necessary for nonphotochemical quenching but not for efficient light harvesting and photosynthesis. These results indicate that PsbS may be the site for nonphotochemical quenching, a finding that has implications for the functional evolution of pigment-binding proteins.
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                Author and article information

                Contributors
                bernard.lepetit@uni-konstanz.de
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                13 September 2019
                13 September 2019
                2019
                : 10
                : 4167
                Affiliations
                [1 ]ISNI 0000 0001 0658 7699, GRID grid.9811.1, Plant Ecophysiology, Department of Biology, , University of Konstanz, ; 78457 Konstanz, Germany
                [2 ]ISNI 0000 0004 1936 8796, GRID grid.430387.b, Environmental Biophysics and Molecular Ecology Program, Department of Marine and Coastal Sciences, Rutgers, , The State University of New Jersey, ; New Brunswick, NJ 08901 USA
                [3 ]ISNI 0000 0001 0726 5157, GRID grid.5734.5, Institute of Genetics, Vetsuisse Faculty, , University of Bern, ; 3001 Bern, Switzerland
                [4 ]ISNI 0000 0004 0459 3739, GRID grid.503320.7, Sorbonne Université, Centre National de la Recherche Scientifique, Institut de Biologie Paris-Seine, , Laboratory of Computational and Quantitative Biology, ; F-75005 Paris, France
                [5 ]ISNI 0000 0004 1936 8390, GRID grid.23856.3a, UMI 3376 Takuvik, CNRS/ULaval, Département de Biologie, Pavillon Alexandre-Vachon, , Université Laval, ; Québec (Québec), G1V 0A6 Canada
                [6 ]ISNI 0000 0001 0658 7699, GRID grid.9811.1, Zukunftskolleg, , University of Konstanz, ; 78457 Konstanz, Germany
                Author information
                http://orcid.org/0000-0002-5644-0182
                http://orcid.org/0000-0002-0584-6748
                http://orcid.org/0000-0003-4734-8955
                http://orcid.org/0000-0001-9980-9210
                Article
                12043
                10.1038/s41467-019-12043-6
                6744471
                31519883
                570cf90f-3883-4e65-9019-73835e5c2728
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 30 March 2019
                : 16 August 2019
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100001711, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Swiss National Science Foundation);
                Award ID: 31003A_172964
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100004794, Centre National de la Recherche Scientifique (National Center for Scientific Research);
                Funded by: FundRef https://doi.org/10.13039/501100002790, Canadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada (NSERC Canadian Network for Research and Innovation in Machining Technology);
                Funded by: FundRef https://doi.org/10.13039/501100001659, Deutsche Forschungsgemeinschaft (German Research Foundation);
                Award ID: KR 1661/8-2
                Award ID: LE 3358/3-1
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100008316, Baden-Württemberg Stiftung (Baden-Württemberg Foundation);
                Award ID: Elite program
                Award Recipient :
                Funded by: -Zukunftskolleg Interdisciplinary Collaborative Project -Marie Curie Zukunftskolleg Incoming Fellowship (grant no. 291784) -Zukunftskolleg Interim grant
                Categories
                Article
                Custom metadata
                © The Author(s) 2019

                Uncategorized
                molecular biology,physiology,non-photochemical quenching
                Uncategorized
                molecular biology, physiology, non-photochemical quenching

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