Photosynthetic organisms harvest light for energy. Some eukaryotic algae have specialized in harvesting far-red light by tuning chlorophyll a absorption through a mechanism still to be elucidated. Here, we combined optically detected magnetic resonance and pulsed electron paramagnetic resonance measurements on red-adapted light-harvesting complexes, rVCP, isolated from the freshwater eustigmatophyte alga Trachydiscus minutus to identify the location of the pigments responsible for this remarkable adaptation. The pigments have been found to belong to an excitonic cluster of chlorophylls a at the core of the complex, close to the central carotenoids in L1/L2 sites. A pair of structural features of the Chl a403/a603 binding site, namely the histidine-to-asparagine substitution in the magnesium-ligation residue and the small size of the amino acid at the i-4 position, resulting in a [A/G]xxxN motif, are proposed to be the origin of this trait. Phylogenetic analysis of various eukaryotic red antennae identified several potential LHCs that could share this tuning mechanism. This knowledge of the red light acclimation mechanism in algae is a step towards rational design of algal strains in order to enhance light capture and efficiency in large-scale biotechnology applications.

• MeSH
• Chlorophyll A * metabolism   chemistry   MeSH
• Chlorophyll metabolism   MeSH
• Electron Spin Resonance Spectroscopy MeSH
• Phylogeny MeSH
• Light MeSH
• Light-Harvesting Protein Complexes * metabolism   genetics   chemistry   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll A * MeSH
• Chlorophyll MeSH
• Light-Harvesting Protein Complexes * MeSH

Antenna proteins play a major role in the regulation of light-harvesting in photosynthesis. However, less is known about a possible link between their sizes (oligomerization state) and fluorescence intensity (number of photons emitted). Here, we used a microscopy-based method, Fluorescence Correlation Spectroscopy (FCS), to analyze different antenna proteins at the particle level. The direct comparison indicated that Chromera Light Harvesting (CLH) antenna particles (isolated from Chromera velia) behaved as the monomeric Light Harvesting Complex II (LHCII) (from higher plants), in terms of their radius (based on the diffusion time) and fluorescence yields. FCS data thus indicated a monomeric oligomerization state of algal CLH antenna (at our experimental conditions) that was later confirmed also by biochemical experiments. Additionally, our data provide a proof of concept that the FCS method is well suited to measure proteins sizes (oligomerization state) and fluorescence intensities (photon counts) of antenna proteins per single particle (monomers and oligomers). We proved that antenna monomers (CLH and LHCIIm) are more "quenched" than the corresponding trimers. The FCS measurement thus represents a useful experimental approach that allows studying the role of antenna oligomerization in the mechanism of photoprotection.

• Keywords
• Chromera velia, antenna proteins, fluorescence correlation spectroscopy, light-harvesting, microscopy, photosynthesis, protein diffusion, protein oligomerization,
• MeSH
• Algal Proteins chemistry   metabolism   MeSH
• Fluorescence * MeSH
• Spectrometry, Fluorescence MeSH
• Photosynthesis * MeSH
• Kinetics MeSH
• Protein Multimerization MeSH
• Protein Transport MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Algal Proteins MeSH

Oxygenic photosynthesis takes place in thylakoid membranes (TM) of cyanobacteria, algae, and higher plants. It begins with light absorption by pigments in large (modular) assemblies of pigment-binding proteins, which then transfer excitation energy to the photosynthetic reaction centers of photosystem (PS) I and PSII. In green algae and plants, these light-harvesting protein complexes contain chlorophylls (Chls) and carotenoids (Cars). However, cyanobacteria, red algae, and glaucophytes contain, in addition, phycobiliproteins in phycobilisomes that are attached to the stromal surface of TM, and transfer excitation energy to the reaction centers via the Chl a molecules in the inner antennas of PSI and PSII. The color and the intensity of the light to which these photosynthetic organisms are exposed in their environment have a great influence on the composition and the structure of the light-harvesting complexes (the antenna) as well as the rest of the photosynthetic apparatus, thus affecting the photosynthetic process and even the entire organism. We present here a perspective on 'Light Quality and Oxygenic Photosynthesis', in memory of George Christos Papageorgiou (9 May 1933-21 November 2020; see notes a and b). Our review includes (1) the influence of the solar spectrum on the antenna composition, and the special significance of Chl a; (2) the effects of light quality on photosynthesis, measured using Chl a fluorescence; and (3) the importance of light quality, intensity, and its duration for the optimal growth of photosynthetic organisms.

• Keywords
• Chl fluorescence induction, chromatic acclimation of cyanobacteria, photoreceptors, photosynthetic pigments, photosystems I and II, stomatal and chloroplast photoinduced movements,
• Publication type
• Journal Article MeSH
• Review MeSH

Survival of phototrophic organisms depends on their ability to collect and convert enough light energy to support their metabolism. Phototrophs can extend their absorption cross section by using diverse pigments and by tuning the properties of these pigments via pigment-pigment and pigment-protein interaction. It is well known that some cyanobacteria can grow in heavily shaded habitats by utilizing far-red light harvested with far-red-absorbing chlorophylls d and f. We describe a red-shifted light-harvesting system based on chlorophyll a from a freshwater eustigmatophyte alga Trachydiscus minutus (Eustigmatophyceae, Goniochloridales). A comprehensive characterization of the photosynthetic apparatus of T. minutus is presented. We show that thylakoid membranes of T. minutus contain light-harvesting complexes of several sizes differing in the relative amount of far-red chlorophyll a forms absorbing around 700 nm. The pigment arrangement of the major red-shifted light-harvesting complex is similar to that of the red-shifted antenna of a marine alveolate alga Chromera velia. Evolutionary aspects of the algal far-red light-harvesting complexes are discussed. The presence of these antennas in eustigmatophyte algae opens up new ways to modify organisms of this promising group for effective use of far-red light in mass cultures.

• Keywords
• Chromatic acclimation, Eustigmatophyta, Light-harvesting protein, Oligomeric LHC, Red-shifted LHC, Violaxanthin,
• MeSH
• Pigments, Biological metabolism   MeSH
• Diuron MeSH
• Spectrometry, Fluorescence MeSH
• Stramenopiles metabolism   radiation effects   MeSH
• Membrane Proteins metabolism   MeSH
• Fresh Water * MeSH
• Light * MeSH
• Light-Harvesting Protein Complexes metabolism   MeSH
• Temperature MeSH
• Thylakoids metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Pigments, Biological MeSH
• Diuron MeSH
• Membrane Proteins MeSH
• Light-Harvesting Protein Complexes MeSH

Chromera velia is a marine photosynthetic relative of human apicomplexan parasites. It has been isolated from coral reefs and is indicted for being involved in symbioses with hermatypic corals. C. velia has been subject to intensive research, but still very little is known of its response to light quality and quantity. Here, we have studied the growth and compositional responses of C. velia to culture under monochromatic light (blue, green or red), at two photon flux densities (PFD, 20 and 100 μmol photons m-2 s-1). Our results show that C. velia growth rate is unaffected by the quality of light, whereas it responds to PFD. However, light quality influenced cell size, which was smaller for cells exposed to blue monochromatic light, regardless of PFD. PFD strongly influenced carbon allocation: at 20 μmol photons m-2 s-1, carbon was mainly allocated into proteins while at 100 μmol photons m-2 s-1, carbon was allocated mainly into carbohydrate and lipid pools. The blue light treatment caused a decrease in the lipids and carbohydrates to proteins and thus suggested to affect nitrogen metabolism in acclimated cells. Whole-cell absorption spectra revealed the existence of red-shifted chlorophyll a antenna not only under red light but in all low PFD treatments. These findings show the ability of C. velia to successfully adapt and thrive in spectrally very different environments of coral reefs.

• MeSH
• Alveolata growth & development   metabolism   radiation effects   MeSH
• Chlorophyll A metabolism   MeSH
• Photosynthesis radiation effects   MeSH
• Light MeSH
• Carbon metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll A MeSH
• Carbon MeSH

We have used time-resolved absorption and fluorescence spectroscopy with nanosecond resolution to study triplet energy transfer from chlorophylls to carotenoids in a protective process that prevents the formation of reactive singlet oxygen. The light-harvesting complexes studied were isolated from Chromera velia, belonging to a group Alveolata, and Xanthonema debile and Nannochloropsis oceanica, both from Stramenopiles. All three light-harvesting complexes are related to fucoxanthin-chlorophyll protein, but contain only chlorophyll a and no chlorophyll c. In addition, they differ in the carotenoid content. This composition of the complexes allowed us to study the quenching of chlorophyll a triplet states by different carotenoids in a comparable environment. The triplet states of chlorophylls bound to the light-harvesting complexes were quenched by carotenoids with an efficiency close to 100%. Carotenoid triplet states were observed to rise with a ~5 ns lifetime and were spectrally and kinetically homogeneous. The triplet states were formed predominantly on the red-most chlorophylls and were quenched by carotenoids which were further identified or at least spectrally characterized.

• Keywords
• Algae, Energy transfer, Light harvesting, Photoprotection, Photosynthesis, Transient spectroscopy,
• MeSH
• Anaerobiosis MeSH
• Time Factors MeSH
• Chlorophyll metabolism   MeSH
• Spectrometry, Fluorescence MeSH
• Photochemical Processes * MeSH
• Stramenopiles metabolism   MeSH
• Carotenoids metabolism   MeSH
• Kinetics MeSH
• Chlorophyll Binding Proteins metabolism   MeSH
• Light-Harvesting Protein Complexes metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll MeSH
• Carotenoids MeSH
• Chlorophyll Binding Proteins MeSH
• Light-Harvesting Protein Complexes MeSH

Diatoms greatly contribute to carbon fixation and thus strongly influence the global biogeochemical balance. Capable of chromatic acclimation (CA) to unfavourable light conditions, diatoms often dominate benthic ecosystems in addition to their planktonic lifestyle. Although CA has been studied at the molecular level, our understanding of this phenomenon remains incomplete. Here we provide new data to better explain the acclimation-associated changes under red-enhanced ambient light (RL) in diatom Phaeodactylum tricornutum, known to express a red-shifted antenna complex (F710). The complex was found to be an oligomer of a single polypeptide, Lhcf15. The steady-state spectroscopic properties of the oligomer were also studied. The oligomeric assembly of the Lhcf15 subunits is required for the complex to exhibit a red-shifted absorption. The presence of the red antenna in RL culture coincides with the development of a rounded phenotype of the diatom cell. A model summarizing the modulation of the photosynthetic apparatus during the acclimation response to light of different spectral quality is proposed. Our study suggests that toggling between alternative organizations of photosynthetic apparatus and distinct cell morphologies underlies the remarkable acclimation capacity of diatoms.

• MeSH
• Phenotype * MeSH
• Adaptation, Physiological MeSH
• Protein Multimerization MeSH
• Diatoms physiology   radiation effects   MeSH
• Spectrum Analysis MeSH
• Light-Harvesting Protein Complexes metabolism   MeSH
• Aquatic Organisms physiology   radiation effects   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Light-Harvesting Protein Complexes MeSH

Photosystem I (PSI) is a multi-subunit integral pigment-protein complex that performs light-driven electron transfer from plastocyanin to ferredoxin in the thylakoid membrane of oxygenic photoautotrophs. In order to achieve the optimal photosynthetic performance under ambient irradiance, the absorption cross section of PSI is extended by means of peripheral antenna complexes. In eukaryotes, this role is played mostly by the pigment-protein complexes of the LHC family. The structure of the PSI-antenna supercomplexes has been relatively well understood in organisms harboring the primary plastid: red algae, green algae and plants. The secondary endosymbiotic algae, despite their major ecological importance, have so far received less attention. Here we report a detailed structural analysis of the antenna-PSI association in the stramenopile alga Nannochloropsis oceanica (Eustigmatophyceae). Several types of PSI-antenna assemblies are identified allowing for identification of antenna docking sites on the PSI core. Instances of departure of the stramenopile system from the red algal model of PSI-Lhcr structure are recorded, and evolutionary implications of these observations are discussed.

• Keywords
• Electron microscopy, Light-harvesting complex, Nannochloropsis, Photosystem I, Stramenopila,
• MeSH
• Photosystem I Protein Complex metabolism   MeSH
• Plastids metabolism   MeSH
• Rhodophyta metabolism   MeSH
• Spectrophotometry, Ultraviolet MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Photosystem I Protein Complex MeSH

Eustigmatophyte algae represent an interesting model system for the study of the regulation of the excitation energy flow due to their use of violaxanthin both as a major light-harvesting pigment and as the basis of xanthophyll cycle. Fluorescence induction kinetics was studied in an oleaginous marine alga Nannochloropsis oceanica. Nonphotochemical fluorescence quenching was analyzed in detail with respect to the state of the cellular xanthophyll pool. Two components of nonphotochemical fluorescence quenching (NPQ), both dependent on the presence of zeaxanthin, were clearly resolved, denoted as slow and fast NPQ based on kinetics of their formation. The slow component was shown to be in direct proportion to the amount of zeaxanthin, while the fast NPQ component was transiently induced in the presence of membrane potential on subsecond timescales. The applicability of these observations to other eustigmatophyte species is demonstrated by measurements of other representatives of this algal group, both marine and freshwater.

• Keywords
• Chl a fluorescence, Eustigmatophyceae, Nannochloropsis, Nonphotochemical quenching, Xanthophyll cycle,
• MeSH
• Fluorescence MeSH
• Photosynthesis MeSH
• Seaweed chemistry   MeSH
• Publication type
• Journal Article MeSH

We present proteomic, spectroscopic, and phylogenetic analysis of light-harvesting protein (Lhc) function in oleaginous Nannochloropsis oceanica (Eustigmatophyta, Stramenopila). N. oceanica utilizes Lhcs of multiple classes: Lhcr-type proteins (related to red algae LHCI), Lhcv (VCP) proteins (violaxanthin-containing Lhcs related to Lhcf/FCP proteins of diatoms), Lhcx proteins (related to Lhcx/LhcSR of diatoms and green algae), and Lhc proteins related to Red-CLH of Chromera velia. Altogether, 17 Lhc-type proteins of the 21 known from genomic data were found in our proteomic analyses. Besides Lhcr-type antennas, a RedCAP protein and a member of the Lhcx protein subfamily were found in association with Photosystem I. The free antenna fraction is formed by trimers of a mixture of Lhcs of varied origins (Lhcv, Lhcr, Lhcx, and relatives of Red-CLH). Despite possessing several proteins of the Red-CLH-type Lhc clade, N. oceanica is not capable of chromatic adaptation under the same conditions as the diatom Phaeodactylum tricornutum or C. velia. In addition, a naming scheme of Nannochloropsis Lhcs is proposed to facilitate further work.

• Keywords
• Light harvesting, Thylakoid membrane, Vaucheriaxanthin, Violaxanthin–chlorophyll protein,
• MeSH
• Photosystem I Protein Complex metabolism   MeSH
• Phylogeny MeSH
• Stramenopiles genetics   metabolism   MeSH
• Spectrophotometry, Ultraviolet MeSH
• Light-Harvesting Protein Complexes chemistry   genetics   metabolism   MeSH
• Tandem Mass Spectrometry MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Photosystem I Protein Complex MeSH
• Light-Harvesting Protein Complexes MeSH