A novel method produces native light-harvesting complex II aggregates from the photosynthetic membrane revealing their role in nonphotochemical quenching.

Nonphotochemical quenching (NPQ) is a mechanism of regulating light harvesting that protects the photosynthetic apparatus from photodamage by dissipating excess absorbed excitation energy as heat. In higher plants, the major light-harvesting antenna complex (LHCII) of photosystem (PS) II is directly involved in NPQ. The aggregation of LHCII is proposed to be involved in quenching. However, the lack of success in isolating native LHCII aggregates has limited the direct interrogation of this process. The isolation of LHCII in its native state from thylakoid membranes has been problematic because of the use of detergent, which tends to dissociate loosely bound proteins, and the abundance of pigment-protein complexes (e.g. PSI and PSII) embedded in the photosynthetic membrane, which hinders the preparation of aggregated LHCII. Here, we used a novel purification method employing detergent and amphipols to entrap LHCII in its natural states. To enrich the photosynthetic membrane with the major LHCII, we used Arabidopsis thaliana plants lacking the PSII minor antenna complexes (NoM), treated with lincomycin to inhibit the synthesis of PSI and PSII core proteins. Using sucrose density gradients, we succeeded in isolating the trimeric and aggregated forms of LHCII antenna. Violaxanthin- and zeaxanthin-enriched complexes were investigated in dark-adapted, NPQ, and dark recovery states. Zeaxanthin-enriched antenna complexes showed the greatest amount of aggregated LHCII. Notably, the amount of aggregated LHCII decreased upon relaxation of NPQ. Employing this novel preparative method, we obtained a direct evidence for the role of in vivo LHCII aggregation in NPQ

Impaired photoprotection in Phaeodactylum tricornutum KEA3 mutants reveals the proton regulatory circuit of diatoms light acclimation

International audienceDiatoms are successful phytoplankton clades able to acclimate to changing environmental conditions, including e.g. variable light intensity. Diatoms are outstanding at dissipating light energy exceeding the maximum photosynthetic electron transfer (PET) capacity via the nonphotochemical quenching (NPQ) process. While the molecular effectors of NPQ as well as the involvement of the proton motive force (PMF) in its regulation are known, the regulators of the PET/PMF relationship remain unidentified in diatoms. We generated mutants of the H$^+$ /K$^+$ antiporter KEA3 in the model diatom $Phaeodactylum\ tricornutum$. Loss of KEA3 activity affects the PET/PMF coupling and NPQ responses at the onset of illumination, during transients and in steady-state conditions. Thus, this antiporter is a main regulator of the PET/PMF coupling. Consistent with this conclusion, a parsimonious model including only two free components, KEA3 and the diadinoxanthin de-epoxidase, describes most of the feedback loops between PET and NPQ. This simple regulatory system allows for efficient responses to fast (minutes) or slow (e.g. diel) changes in light environment, thanks to the presence of a regulatory calcium ion (Ca$^{2+}$ )-binding domain in KEA3 modulating its activity. This circuit is likely tuned by the NPQ-effector proteins, LHCXs, providing diatoms with the required flexibility to thrive in different ocean provinces

Discerning the effects of photoinhibition and photoprotection on the rate of oxygen evolution in Arabidopsis leaves

This work was supported by the Leverhulme Trust grant RPG-2012-478 and UK Biotechnology and Biological Sciences Research Council grant BB/L019027/1 to AVR. AVR would like to acknowledge The Royal Society for the Wolfson Research Merit Award

PsbS protein modulates non-photochemical chlorophyll fluorescence quenching in membranes depleted of photosystems

© 2015 Elsevier B.V. All rights reserved. Plants with varying levels of PsbS protein were grown on lincomycin. Enhanced levels of non-photochemical fluorescence quenching (NPQ) in over-expressers of the protein have been observed. This was accompanied by increased amplitude of the irreversible NPQ component, qI, previously considered to reflect mainly photoinhibition of PSII reaction centres (RCII). However, since RCIIs were largely absent the observed qI is likely to originate from the LHCII antenna. In chloroplasts of over-expressers of PsbS grown on lincomycin an abnormally large NPQ (∼7) was characterised by a 0.34 ns average chlorophyll fluorescence lifetime. Yet the lifetime in the Fm state was similar to that of wild-type plants. 77 K fluorescence emission spectra revealed a specific 700 nm peak typical of LHCII aggregates as well as quenching of the PSI fluorescence at 730 nm. The aggregated state manifested itself as a clear change in the distance between LHCII complexes detected by freeze-fracture electron microscopy. Grana thylakoids in the quenched state revealed 3 times more aggregated LHCII particles compared to the dark-adapted state. Overall, the results directly demonstrate the importance of LHCII aggregation in the NPQ mechanism and show that the PSII supercomplex structure plays no role in formation of the observed quenching.This work was supported by Queen Mary Principal’s research studentship to MAW and The Leverhulme Trust and UK Biotechnology and Biological Sciences Research Council to AVR. AVR would like to acknowledge The Royal Society for the Wolfson Research Merit Award

Spectral radiation dependent photoprotective mechanism in the diatom Pseudo-nitzschia multistriata.

Phytoplankton, such as diatoms, experience great variations of photon flux density (PFD) and light spectrum along the marine water column. Diatoms have developed some rapidly-regulated photoprotective mechanisms, such as the xanthophyll cycle activation (XC) and the non-photochemical chlorophyll fluorescence quenching (NPQ), to protect themselves from photooxidative damages caused by excess PFD. In this study, we investigate the role of blue fluence rate in combination with red radiation in shaping photoacclimative and protective responses in the coastal diatom Pseudo-nitzschia multistriata. This diatom was acclimated to four spectral light conditions (blue, red, blue-red, blue-red-green), each of them provided with low and high PFD. Our results reveal that the increase in the XC pool size and the amplitude of NPQ is determined by the blue fluence rate experienced by cells, while cells require sensing red radiation to allow the development of these processes. Variations in the light spectrum and in the blue versus red radiation modulate either the photoprotective capacity, such as the activation of the diadinoxanthin-diatoxanthin xanthophyll cycle, the diadinoxanthin de-epoxidation rate and the capacity of non-photochemical quenching, or the pigment composition of this diatom. We propose that spectral composition of light has a key role on the ability of diatoms to finely balance light harvesting and photoprotective capacity

Assessing the role of dust deposition on phytoplankton ecophysiology and succession in a low-nutrient low-chlorophyll ecosystem: a mesocosm experiment in the Mediterranean Sea

International audienceIn this study, we investigate the response of the phytoplankton community, with emphasis on ecophysiology and succession, after two experimental additions of Saharan dust in the surface water layer of a low-nutrient low-chlorophyll ecosystem in the Mediterranean Sea. Three mesocosms were amended with evapocondensed dust to simulate realistic Saharan dust events, while three additional mesocosms were kept unamended and served as controls. The experiment consisted in two consecutive dust additions and samples were daily collected at different depths (-0.1, -5 and -10 m) during one week, starting before each addition occurred. Data concerning HPLC pigment analysis on two size classes ( 3 μm), electron transport rate (ETR) vs. irradiance curves, non-photochemical fluorescence quenching (NPQ) and phytoplankton cell abundance (measured by flow cytometry), are presented and discussed in this paper. Results show that picophytoplankton mainly respond to the first dust addition, while the second addition leads to an increase of both pico- and nano-/microphytoplankton. Ecophysiological changes in the phytoplankton community occur, with NPQ and pigment concentration per cell increasing after dust additions. While biomass increases after pulses of new nutrients, ETR does not greatly vary between dust-amended and control conditions, in relation with ecophysiological changes within the phytoplankton community, such as the increase in NPQ and pigment cellular concentration. A quantitative assessment and parameterisation of the onset of a phytoplankton bloom in a nutrient-limited ecosystem is attempted on the basis of the increase in phytoplankton biomass observed during the experiment. The results of this study are discussed focusing on the adaptation of picophytoplankton to nutrient limitation in the surface water layer, as well as on size-dependent competition ability in phytoplankton