Photosynthesis Research (v.123, #1)

ATP is synthesized using ATP synthase by utilizing energy either from the oxidation of organic compounds, or from light, via redox reactions (oxidative- or photo phosphorylation), in energy-transforming membranes of mitochondria, chloroplasts, and bacteria. ATP synthase undergoes several changes during its functioning. The generally accepted model for ATP synthesis is the well-known rotatory model (see e.g., Junge et al., Nature 459:364–370, 2009; Junge and Müller, Science 333:704–705, 2011). Here, we present an alternative modified model for the coupling of electron and proton transfer to ATP synthesis, which was initially developed by Albert Lester Lehninger (1917–1986). Details of the molecular mechanism of ATP synthesis are described here that involves cyclic low-amplitude shrinkage and swelling of mitochondria. A comparison of the well-known current model and the mechano-chemiosmotic model is also presented. Based on structural, and other data, we suggest that ATP synthase is a Ca2+/H+–K+ Cl–pump–pore–enzyme complex, in which γ-subunit rotates 360° in steps of 30°, and 90° due to the binding of phosphate ions to positively charged amino acid residues in the N-terminal γ-subunit, while in the electric field. The coiled coil b 2-subunits are suggested to act as ropes that are shortened by binding of phosphate ions to positively charged lysines or arginines; this process is suggested to pull the α 3 β 3-hexamer to the membrane during the energization process. ATP is then synthesized during the reverse rotation of the γ-subunit by destabilizing the phosphated N-terminal γ-subunit and b 2-subunits under the influence of Ca2+ ions, which are pumped over from storage—intermembrane space into the matrix, during swelling of intermembrane space. In the process of ATP synthesis, energy is first, predominantly, used in the delivery of phosphate ions and protons to the α 3 β 3-hexamer against the energy barrier with the help of C-terminal alpha-helix of γ-subunit that acts as a lift; then, in the formation of phosphoryl group; and lastly, in the release of ATP molecules from the active center of the enzyme and the loading of ADP. We are aware that our model is not an accepted model for ATP synthesis, but it is presented here for further examination and test.
Keywords: F0F1-ATPase; γ-Subunit rotation; Mechano-chemiosmotic model; Shrinkage–swelling; Electron transfer

The origin of the split B800 absorption peak in the LH2 complexes from Allochromatium vinosum by Alexander Löhner; Anne-Marie Carey; Kirsty Hacking; Nichola Picken; Sharon Kelly; Richard Cogdell; Jürgen Köhler (23-31).
The absorption spectrum of the high-light peripheral light-harvesting (LH) complex from the photosynthetic purple bacterium Allochromatium vinosum features two strong absorptions around 800 and 850 nm. For the LH2 complexes from the species Rhodopseudomonas acidophila and Rhodospirillum molischianum, where high-resolution X-ray structures are available, similar bands have been observed and were assigned to two pigment pools of BChl a molecules that are arranged in two concentric rings (B800 and B850) with nine (acidophila) or eight (molischianum) repeat units, respectively. However, for the high-light peripheral LH complex from Alc. vinosum, the intruiging feature is that the B800 band is split into two components. We have studied this pigment–protein complex by ensemble CD spectroscopy and polarisation-resolved single-molecule spectroscopy. Assuming that the high-light peripheral LH complex in Alc. vinosum is constructed on the same modular principle as described for LH2 from Rps. acidophila and Rsp. molischianum, we used those repeat units as a starting point for simulating the spectra. We find the best agreement between simulation and experiment for a ring-like oligomer of 12 repeat units, where the mutual arrangement of the B800 and B850 rings resembles those from Rsp. molischianum. The splitting of the B800 band can be reproduced if both an excitonic coupling between dimers of B800 molecules and their interaction with the B850 manifold are taken into account. Such dimers predict an interesting apoprotein organisation as discussed below.
Keywords: Light-harvesting complexes; Allochromatium vinosum ; Single-molecule spectroscopy

This study identifies Salsola laricifolia as a C3–C4 intermediate in tribe Salsoleae s.l., Chenopodiaceae, and compares S. laricifolia with the previously described C3–C4 intermediates in Salsoleae. Photosynthetic pathway characteristics were studied in four species of this tribe including S. laricifolia, C3 Sympegma regelii, C3–C4 S. arbusculiformis, and C4 S. arbuscula, using the approaches of leaf anatomy and ultrastructure, activities of ribulose 1-5-bisphosphate carboxylase/oxygenase (Rubisco) and PEP carboxylase (PEPC), CO2 compensation point, and immunolocalization of Rubisco, PEPC, and the P-subunit of glycine decarboxylase (GDC). Salsola laricifolia has intermediate features, with near continuous and distinctive Kranz-like cells (KLCs) compared with the C3-Sympegmoid anatomical type and the C3–C4 intermediate S. arbusculiformis, a relatively low CO2 compensation point (30.4 μmol mol−1) and mesophyll (M)-to KLC tissue ratio, mitochondria in KLCs primarily occurring along the centripetal wall, and specific localization of P-protein GDC in the KLCs. The C3-type isotope value (−22.4 ‰), the absence of the clear labeling for PEPC in M cells, and the low activity of the PEPC enzyme (61.5 μmol mg−1 chlorophyll−1 h−1) support the identification of S. laricifolia as a type I C3–C4 intermediate. Although these C3–C4 intermediate species have different structural features, one with discontinuous KL cells and the other with continuous, they have similar characteristics in physiology and biochemistry.
Keywords: C3–C4 intermediate species; Kranz anatomy; Chenopodiaceae; C4 photosynthesis; Photosynthetic enzymes; Salsola laricifolia

The Prochlorococcus carbon dioxide-concentrating mechanism: evidence of carboxysome-associated heterogeneity by Claire S. Ting; Katharine H. Dusenbury; Reid A. Pryzant; Kathleen W. Higgins; Catherine J. Pang; Christie E. Black; Ellen M. Beauchamp (45-60).
The ability of Prochlorococcus to numerically dominate open ocean regions and contribute significantly to global carbon cycles is dependent in large part on its effectiveness in transforming light energy into compounds used in cell growth, maintenance, and division. Integral to these processes is the carbon dioxide-concentrating mechanism (CCM), which enhances photosynthetic CO2 fixation. The CCM involves both active uptake systems that permit intracellular accumulation of inorganic carbon as the pool of bicarbonate and the system of HCO3 conversion into CO2. The latter is located in the carboxysome, a microcompartment designed to promote the carboxylase activity of Rubisco. This study presents a comparative analysis of several facets of the Prochlorococcus CCM. Our analyses indicate that a core set of CCM components is shared, and their genomic organization is relatively well conserved. Moreover, certain elements, including carboxysome shell polypeptides CsoS1 and CsoS4A, exhibit striking conservation. Unexpectedly, our analyses reveal that the carbonic anhydrase (CsoSCA) and CsoS2 shell polypeptide have diversified within the lineage. Differences in csoSCA and csoS2 are consistent with a model of unequal rates of evolution rather than relaxed selection. The csoS2 and csoSCA genes form a cluster in Prochlorococcus genomes, and we identified two conserved motifs directly upstream of this cluster that differ from the motif in marine Synechococcus and could be involved in regulation of gene expression. Although several elements of the CCM remain well conserved in the Prochlorococcus lineage, the evolution of differences in specific carboxysome features could in part reflect optimization of carboxysome-associated processes in dissimilar cellular environments.
Keywords: Chlorophyll b-containing cyanobacteria; Genomic diversity; Global comparative genomics; Oxychlorobacteria; Microcompartment; Carbonic anhydrase

Isolation and characterization of a PSI–LHCI super-complex and its sub-complexes from a siphonaceous marine green alga, Bryopsis Corticulans by Xiaochun Qin; Wenda Wang; Lijing Chang; Jinghua Chen; Peng Wang; Jianping Zhang; Yikun He; Tingyun Kuang; Jian-Ren Shen (61-76).
A novel super-complex of photosystem I (PSI)–light-harvesting complex I (LHCI) was isolated from a siphonaceous marine green alga, Bryopsis corticulans. The super-complex contained 9–10 Lhca antennas as external LHCI bound to the core complex. The super-complex was further disintegrated into PSI core and LHCI sub-complexes, and analysis of the pigment compositions by high-performance liquid chromatography revealed unique characteristics of the B. corticulans PSI in that one PSI core contained around 14 α-carotenes and 1–2 ε-carotenes. This is in sharp contrast to the PSI core from higher plants and most cyanobacteria where only β-carotenes were present, and is the first report for an α-carotene-type PSI core complex among photosynthetic eukaryotes, suggesting a structural flexibility of the PSI core. Lhca antennas from B. corticulans contained seven kinds of carotenoids (siphonaxanthin, all-trans neoxanthin, 9′-cis neoxanthin, violaxanthin, siphonein, ε-carotene, and α-carotene) and showed a high carotenoid:chlorophyll ratio of around 7.5:13. PSI–LHCI super-complex and PSI core showed fluorescence emission peaks at 716 and 718 nm at 77 K, respectively; whereas two Lhca oligomers had fluorescence peaks at 681 and 684 nm, respectively. By comparison with spinach PSI preparations, it was found that B. corticulans PSI had less red chlorophylls, most of them are present in the core complex but not in the outer light-harvesting systems. These characteristics may contribute to the fine tuning of the energy transfer network, and to acclimate to the ever-changing light conditions under which the unique green alga inhabits.
Keywords: Bryopsis corticulans ; α-Carotene; Siphonaxanthin; Siphonein; Neoxanthin; Photosystem I

Theoretical prediction of effective mean PAR in optically dense samples is complicated by various optical effects, including light scattering and reflections. Direct information on the mean rate of photon absorption by PS II is provided by the kinetics of the fluorescence rise induced upon onset of strong actinic illumination (O-I1 rise). A recently introduced kinetic multi-color PAM fluorometer was applied to study the relationship between initial slope and cell density in the relatively simple model system of suspensions of Chlorella. Use of a curve fitting routine was made which was originally developed for assessment of the wavelength-dependent absorption cross-section of PS II, σ II(λ), in dilute suspensions. The model underlying analysis of the O-I1 rise kinetics is outlined and data on the relationship between fitted values of σ II(λ) and PAR in dilute samples are presented. With increasing cell density, lowering of apparent cross-section, <σ>(λ), with respect to σ II(λ), relates to a decrease of effective mean PAR, (λ), relative to incident PAR(λ). When ML and AL are applied in the same direction, the decline of <σ>(λ)/σ II(λ) with increasing optical density is less steep than that of the theoretically predicted (λ)/PAR(λ). It approaches a value of 0.5 when the same colors of ML and AL are used, in agreement with theory. These observations open the way for estimating mean PAR in optically dense samples via measurements of <σ>(λ)/σ II(λ)).
Keywords: Absorption cross-section of PS II; Chlorella ; Chlorophyll fluorescence; Multi-color-PAM; O-I1 rise kinetics; Photosynthesis; Polyphasic fluorescence rise

Photophysiology and daily primary production of a temperate symbiotic gorgonian by C. Ferrier-Pagès; S. Reynaud; E. Béraud; C. Rottier; D. Menu; G. Duong; F. Gévaert (95-104).
Gorgonians are one of the most important benthic components of tropical and temperate areas, and play a fundamental role as ecosystem engineers. Although global warming and pollution increasingly threaten them, the acquisition of nutrients, which is a key process in fitness and stress resistance, has been poorly investigated in such species. This study has thus used an advanced in situ incubation chamber for the first time with gorgonians, to assess the daily acquisition of nutrients and the photophysiology of the Mediterranean symbiotic species, Eunicella singularis. The xanthophyll cycle was assessed in parallel. This work has revealed that E. singularis presents a different functioning than the Mediterranean symbiotic corals. This gorgonian indeed relies on both autotrophy and heterotrophy in summer to optimize its energetic budget, while corals mainly shift to autotrophy for their respiratory needs and tissue growth. In addition, although E. singularis lives in the same depths/locations, and harbours the same symbiont genotype than the corals, the photosynthetic performances of their respective symbionts are significantly different. Indeed, E. singularis acquired 2–3 times less autotrophic carbon from its symbionts than corals, but maintained a positive carbon budget by reducing respiration rates, and by presenting maximal photosynthetic rates throughout the day, suggesting a very efficient light utilization. Almost no photoinhibition was observed under very high light levels, because of the induction of a xanthophyll photoprotection process. These results help understanding why gorgonians often dominate many benthic ecosystems.
Keywords: Temperate gorgonians; Symbionts; Photosynthesis; Carbon budget; Xanthophyll cycle; Photoacclimation

Teaching the Z-Scheme of electron transport in photosynthesis: a perspective by Pradipta Kumar Mohapatra; Nihar Ranjan Singh (105-114).
This paper deals with how Govindjee taught the Z-Scheme of electron transport in oxygenic photosynthesis at Ravenshaw University, Cuttack, Odisha, India, in 2014, in a unique and highly effective fashion—using students to act as molecules, representing the entire electron transport chain from water to nicotinamide adenine dinucleotide phosphate (NADP+). It culminated in a show by B.Sc. students in the garden of the Department of Botany, Ravenshaw University. The first author (PKM) personally acted as Ferredoxin NADP Reductase (FNR) catalyzing the reduction of NADP+ to NADPH, taking electrons from reduced ferredoxin at the end of Photosystem I. On the other hand, the Q-cycle was played by M.Sc. students, who acted as molecules running this ingenious cycle that produces extra protons. An interesting event was when a student, acting as a herbicide, who was dressed like a devil (fierce looking, in black clothes with a sword; “Yamaraj: The God of Death”, as he called himself), stopped all reactions by throwing out QB, the second plastoquinone molecule of Photosystem II, and that too aggressively, taking its position instead. The second author was the major organizer of the Z-scheme show. We provide here a basic background on the process, a bit on Govindjee’s teaching, and some selected pictures from the drama played in March, 2014 at Ravenshaw University. Here, we also recognize the teacher Govindjee for his ingenious and fun-filled teaching methods that touched the hearts and the souls of the students as well as the teachers of Ravenshaw University. He was rated as one of the most-admired teachers of plant biology at our university.
Keywords: Electron transport; Govindjee; Photosynthesis; Q-cycle; Z-scheme drama