Oxygenic Photosynthesis in Far-Red Light: Strategies and Mechanisms.

Oxygenic photosynthesis, the process that converts light energy into chemical energy, is traditionally associated with the absorption of visible light by chlorophyll molecules. However, recent studies have revealed a growing number of organisms capable of using far-red light (700-800 nm) to drive oxygenic photosynthesis. This phenomenon challenges the conventional understanding of the limits of this process. In this review, we briefly introduce the organisms that exhibit far-red photosynthesis and explore the different strategies they employ to harvest far-red light. We discuss the modifications of photosynthetic complexes and their impact on the delivery of excitation energy to photochemical centers and on overall photochemical efficiency. Finally, we examine the solutions employed to drive electron transport and water oxidation using relatively low-energy photons. The findings discussed here not only expand our knowledge of the remarkable adaptation capacities of photosynthetic organisms but also offer insights into the potential for enhancing light capture in crops. Expected final online publication date for the Annual Review of Physical Chemistry, Volume 75 is April 2024. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Tenfold sensitivity increase in streak camera detection by propagation synchronous integration without compromising time resolution.

For many applications that involve measuring ultrafast optical phenomena, the streak camera is the device of choice because of its excellent time resolution, its high sensitivity, the possibility to simultaneously measure lifetimes and spectra, and because it can capture the temporal dynamics in a single shot. Nevertheless, to obtain a good time resolution, often narrow slits have to be employed that restrict the image source area and, therefore, limit the light collection efficiency in the experiment. For some applications, it is therefore challenging to find an acceptable balance between the time resolution and signal-to-noise ratio. To overcome this limitation, we have devised the propagation synchronous integration principle for the streak camera, in which an effective spatio-dependent time-shift in the excitation of a sample is introduced and counteracted by the streak sweep, thereby effectively allowing for an increased image source area while maintaining the optimal time resolution. Using the Optronis streak camera with tunable streak sweep and large (1 mm) photocathode width, we could achieve a sevenfold increase in light collection efficiency without affecting the time resolution. Furthermore, we were also able to achieve an 11-fold increase in light collection at the cost of a 26% decrease in the time resolution.

Coloring Outside the Lines: Exploiting Pigment-Protein Synergy for Far-Red Absorption in Plant Light-Harvesting Complexes.

Plants are designed to utilize visible light for photosynthesis. Expanding this light absorption toward the far-red could boost growth in low-light conditions and potentially increase crop productivity in dense canopies. A promising strategy is broadening the absorption of antenna complexes to the far-red. In this study, we investigated the capacity of the photosystem I antenna protein Lhca4 to incorporate far-red absorbing chlorophylls d and f and optimize their spectra. We demonstrate that these pigments can successfully bind to Lhca4, with the protein environment further red-shifting the chlorophyll d absorption, markedly extending the absorption range of this complex above 750 nm. Notably, chlorophyll d substitutes the canonical chlorophyll a red-forms, resulting in the most red-shifted emission observed in a plant light-harvesting complex. Using ultrafast spectroscopy, we show that the introduction of these novel chlorophylls does not interfere with the excited state decay or the energy equilibration processes within the complex. The results demonstrate the feasibility of engineering plant antennae to absorb deeper into the far-red region while preserving their functional and structural integrity, paving the way for innovative strategies to enhance photosynthesis.

Structural comparison of allophycocyanin variants reveals the molecular basis for their spectral differences.

Allophycocyanins are phycobiliproteins that absorb red light and transfer the energy to the reaction centers of oxygenic photosynthesis in cyanobacteria and red algae. Recently, it was shown that some allophycocyanins absorb far-red light and that one subset of these allophycocyanins, comprising subunits from the ApcD4 and ApcB3 subfamilies (FRL-AP), form helical nanotubes. The lowest energy absorbance maximum of the oligomeric ApcD4-ApcB3 complexes occurs at 709 nm, which is unlike allophycocyanin (AP; ApcA-ApcB) and allophycocyanin B (AP-B; ApcD-ApcB) trimers that absorb maximally at ~ 650 nm and ~ 670 nm, respectively. The molecular bases of the different spectra of AP variants are presently unclear. To address this, we structurally compared FRL-AP with AP and AP-B, performed spectroscopic analyses on FRL-AP, and leveraged computational approaches. We show that among AP variants, the α-subunit constrains pyrrole ring A of its phycocyanobilin chromophore to different extents, and the coplanarity of ring A with rings B and C sets a baseline for the absorbance maximum of the chromophore. Upon oligomerization, the α-chromophores of all AP variants exhibit a red shift of the absorbance maximum of ~ 25 to 30 nm and band narrowing. We exclude excitonic coupling in FRL-AP as the basis for this red shift and extend the results to discuss AP and AP-B. Instead, we attribute these spectral changes to a conformational alteration of pyrrole ring D, which becomes more coplanar with rings B and C upon oligomerization. This study expands the molecular understanding of light-harvesting attributes of phycobiliproteins and will aid in designing phycobiliproteins for biotechnological applications.

Drought affects both photosystems in Arabidopsis thaliana.

Drought is a major abiotic stress that impairs plant growth and development. Despite this, a comprehensive understanding of drought effects on the photosynthetic apparatus is lacking. In this work, we studied the consequences of 14-d drought treatment on Arabidopsis thaliana. We used biochemical and spectroscopic methods to examine photosynthetic membrane composition and functionality. Drought led to the disassembly of PSII supercomplexes and the degradation of PSII core. The light-harvesting complexes (LHCII) instead remain in the membrane but cannot act as an antenna for active PSII, thus representing a potential source of photodamage. This effect can also be observed during nonphotochemical quenching (NPQ) induction when even short pulses of saturating light can lead to photoinhibition. At a later stage, under severe drought stress, the PSI antenna size is also reduced and the PSI-LHCI supercomplexes disassemble. Surprisingly, although we did not observe changes in the PSI core protein content, the functionality of PSI is severely affected, suggesting the accumulation of nonfunctional PSI complexes. We conclude that drought affects both photosystems, although at a different stage, and that the operative quantum efficiency of PSII (ΦPSII ) is very sensitive to drought and can thus be used as a parameter for early detection of drought stress.

At the origin of the selectivity of the chlorophyll-binding sites in Light Harvesting Complex II (LHCII).

The photosynthetic light-harvesting complexes (LHCs) are responsible for light absorption due to their pigment-binding properties. These pigments are primarily Chlorophyll (Chl) molecules of type a and b, which ensure an excellent coverage of the visible light spectrum. To date, it is unclear which factors drive the selective binding of different Chl types in the LHC binding pockets. To gain insights into this, we employed molecular dynamics simulations on LHCII binding different Chl types. From the resulting trajectories, we have calculated the binding affinities per each Chl-binding pocket using the Molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) model. To further examine the importance of the nature of the axial ligand in tuning the Chl selectivity of the binding sites, we used Density Functional Theory (DFT) calculations. The results indicate that some binding pockets have a clear Chl selectivity, and the factors governing these selectivities are identified. Other binding pockets are promiscuous, which is consistent with previous in vitro reconstitution studies. DFT calculations show that the nature of the axial ligand is not a major factor in determining the Chl binding pocket selectivity, which is instead probably controlled by the folding process.

Helical allophycocyanin nanotubes absorb far-red light in a thermophilic cyanobacterium.

To compete in certain low-light environments, some cyanobacteria express a paralog of the light-harvesting phycobiliprotein, allophycocyanin (AP), that strongly absorbs far-red light (FRL). Using cryo-electron microscopy and time-resolved absorption spectroscopy, we reveal the structure-function relationship of this FRL-absorbing AP complex (FRL-AP) that is expressed during acclimation to low light and that likely associates with chlorophyll a-containing photosystem I. FRL-AP assembles as helical nanotubes rather than typical toroids due to alterations of the domain geometry within each subunit. Spectroscopic characterization suggests that FRL-AP nanotubes are somewhat inefficient antenna; however, the enhanced ability to harvest FRL when visible light is severely attenuated represents a beneficial trade-off. The results expand the known diversity of light-harvesting proteins in nature and exemplify how biological plasticity is achieved by balancing resource accessibility with efficiency.

The photosynthetic apparatus of the CAM plant Tillandsia flabellate and its response to water deficit.

CAM plants are superior to C3 plants in drought resistance because of their peculiar photosynthesis pathway and morphological features. While those aspects have been studied for decades, little is known about the photosynthetic machinery of CAM plants. Here, we used a combination of biochemical and biophysical methods to study the photosynthetic apparatus of Tillandsia flabellate, an obligatory CAM plant. Most of the Photosystems super- and sub-complexes have properties very similar to those of Arabidopsis, with the main difference that in Tillandsia PSI-LHCI complexes bind extra LHCI. Functional measurements show that the PSI/PSII ratio is rather low compared to other plants and that the antenna size of both PSI and PSII is small. Upon 30-day water deficiency, the composition of the photosystems does not change significantly, PSII efficiency remains high and no Photosystem II photoinhibition was detected despite a reduction of non-photochemical quenching (NPQ).

The origin of pigment-binding differences in CP29 and LHCII: the role of protein structure and dynamics.

The first step of photosynthesis in plants is performed by the light-harvesting complexes (LHC), a large family of pigment-binding proteins embedded in the photosynthetic membranes. These complexes are conserved across species, suggesting that each has a distinct role. However, they display a high degree of sequence homology and their static structures are almost identical. What are then the structural features that determine their different properties? In this work, we compared the two best-characterized LHCs of plants: LHCII and CP29. Using molecular dynamics simulations, we could rationalize the difference between them in terms of pigment-binding properties. The data also show that while the loops between the helices are very flexible, the structure of the transmembrane regions remains very similar in the crystal and the membranes. However, the small structural differences significantly affect the excitonic coupling between some pigment pairs. Finally, we analyzed in detail the structure of the long N-terminus of CP29, showing that it is structurally stable and it remains on top of the membrane even in the absence of other proteins. Although the structural changes upon phosphorylation are minor, they can explain the differences in the absorption properties of the pigments observed experimentally.

Harvesting Far-Red Light with Plant Antenna Complexes Incorporating Chlorophyll d.

Increasing the absorption cross section of plants by introducing far-red absorbing chlorophylls (Chls) has been proposed as a strategy to boost crop yields. To make this strategy effective, these Chls should bind to the photosynthetic complexes without altering their functional architecture. To investigate if plant-specific antenna complexes can provide the protein scaffold to accommodate these Chls, we have reconstituted the main light-harvesting complex (LHC) of plants LHCII in vitro and in silico, with Chl d. The results demonstrate that LHCII can bind Chl d in a number of binding sites, shifting the maximum absorption ∼25 nm toward the red with respect to the wild-type complex (LHCII with Chl a and b) while maintaining the native LHC architecture. Ultrafast spectroscopic measurements show that the complex is functional in light harvesting and excitation energy transfer. Overall, we here demonstrate that it is possible to obtain plant LHCs with enhanced far-red absorption and intact functional properties.

Uncovering the interactions driving carotenoid binding in light-harvesting complexes.

Carotenoids are essential constituents of plant light-harvesting complexes (LHCs), being involved in protein stability, light harvesting, and photoprotection. Unlike chlorophylls, whose binding to LHCs is known to require coordination of the central magnesium, carotenoid binding relies on weaker intermolecular interactions (such as hydrogen bonds and van der Waals forces), whose character is far more elusive. Here we addressed the key interactions responsible for carotenoid binding to LHCs by combining molecular dynamics simulations and polarizable quantum mechanics/molecular mechanics calculations on the major LHC, LHCII. We found that carotenoid binding is mainly stabilized by van der Waals interactions with the surrounding chlorophyll macrocycles rather than by hydrogen bonds to the protein, the latter being more labile than predicted from structural data. Furthermore, the interaction network in the binding pockets is relatively insensitive to the chemical structure of the embedded carotenoid. Our results are consistent with a number of experimental data and challenge the role played by specific interactions in the assembly of pigment-protein complexes.