Photosynthesis Research (v.131, #1)

Influence of light and nitrogen on the photosynthetic efficiency in the C4 plant Miscanthus × giganteus by Jian-Ying Ma; Wei Sun; Nuria K. Koteyeva; Elena Voznesenskaya; Samantha S. Stutz; Anthony Gandin; Andreia M. Smith-Moritz; Joshua L. Heazlewood; Asaph B. Cousins (1-13).
There are numerous studies describing how growth conditions influence the efficiency of C4 photosynthesis. However, it remains unclear how changes in the biochemical capacity versus leaf anatomy drives this acclimation. Therefore, the aim of this study was to determine how growth light and nitrogen availability influence leaf anatomy, biochemistry and the efficiency of the CO2 concentrating mechanism in Miscanthus × giganteus. There was an increase in the mesophyll cell wall surface area but not cell well thickness in the high-light (HL) compared to the low-light (LL) grown plants suggesting a higher mesophyll conductance in the HL plants, which also had greater photosynthetic capacity. Additionally, the HL plants had greater surface area and thickness of bundle-sheath cell walls compared to LL plants, suggesting limited differences in bundle-sheath CO2 conductance because the increased area was offset by thicker cell walls. The gas exchange estimates of phosphoenolpyruvate carboxylase (PEPc) activity were significantly less than the in vitro PEPc activity, suggesting limited substrate availability in the leaf due to low mesophyll CO2 conductance. Finally, leakiness was similar across all growth conditions and generally did not change under the different measurement light conditions. However, differences in the stable isotope composition of leaf material did not correlate with leakiness indicating that dry matter isotope measurements are not a good proxy for leakiness. Taken together, these data suggest that the CO2 concentrating mechanism in Miscanthus is robust under low-light and limited nitrogen growth conditions, and that the observed changes in leaf anatomy and biochemistry likely help to maintain this efficiency.
Keywords: Carbon isotope discrimination; C4 photosynthesis; Miscanthus; Nitrogen; Light

The plastoquinol–plastoquinone exchange mechanism in photosystem II: insight from molecular dynamics simulations by Veranika Zobnina; Maya D. Lambreva; Giuseppina Rea; Gaetano Campi; Amina Antonacci; Viviana Scognamiglio; Maria Teresa Giardi; Fabio Polticelli (15-30).
In the photosystem II (PSII) of oxygenic photosynthetic organisms, the reaction center (RC) core mediates the light-induced electron transfer leading to water splitting and production of reduced plastoquinone molecules. The reduction of plastoquinone to plastoquinol lowers PSII affinity for the latter and leads to its release. However, little is known about the role of protein dynamics in this process. Here, molecular dynamics simulations of the complete PSII complex embedded in a lipid bilayer have been used to investigate the plastoquinol release mechanism. A distinct dynamic behavior of PSII in the presence of plastoquinol is observed which, coupled to changes in charge distribution and electrostatic interactions, causes disruption of the interactions seen in the PSII–plastoquinone complex and leads to the “squeezing out” of plastoquinol from the binding pocket. Displacement of plastoquinol closes the second water channel, recently described in a 2.9 Å resolution PSII structure (Guskov et al. in Nat Struct Mol Biol 16:334–342, 2009), allowing to rule out the proposed “alternating” mechanism of plastoquinol–plastoquinone exchange, while giving support to the “single-channel” one. The performed simulations indicated a pivotal role of D1-Ser264 in modulating the dynamics of the plastoquinone binding pocket and plastoquinol–plastoquinone exchange via its interaction with D1-His252 residue. The effects of the disruption of this hydrogen bond network on the PSII redox reactions were experimentally assessed in the D1 site-directed mutant Ser264Lys.
Keywords: Photosystem II; plastoquinone; Plastoquinol; Protein dynamics; Molecular dynamics simulations; Plastoquinol release mechanism

Metabolism of xenobiotics by Chlamydomonas reinhardtii: Phenol degradation under conditions affecting photosynthesis by Theocharis T. Nazos; Emmanouel J. Kokarakis; Demetrios F. Ghanotakis (31-40).
In the present work, the biodegradation of phenol by axenic cultures of the unicellular microalga Chlamydomonas reinhardtii was investigated. Biodegradation proved to be a dynamic bioenergetic process, affected by changes in the culture conditions. Microalgae biodegraded defined amounts of phenol, as a result of the induced stress caused at high concentrations, despite the fact that this process proved to be energy demanding and thus affected growth of the culture. High levels of biodegradation were observed both in the absence of an alternative carbon source and in the presence of acetate as a carbon source. Biodegradation of phenol by Chlamydomonas proved to be an aerobic, photoregulated process. This is the first time that Chlamydomonas reinhardtii has been used for bioremediation purposes. This study has demonstrated that the most important factor in the biodegradation of phenol is the selection of the appropriate culture conditions (presence or absence of alternative carbon source, light intensity, and oxygen availability) that provide the best bioenergetic balance among growth, induced stress, and biodegradation of phenol.
Keywords: Chlamydomonas reinhardtii ; Phenol; Biodegradation; Photosynthesis; Bioenergetics; Stress

Essential role of the PSI–LHCII supercomplex in photosystem acclimation to light and/or heat conditions by state transitions by Yoko Marutani; Yasuo Yamauchi; Mari Higashiyama; Akihito Miyoshi; Seiji Akimoto; Kanako Inoue; Ken-ichi Ikeda; Masaharu Mizutani; Yukihiro Sugimoto (41-50).
Light and temperature affect state transitions through changes in the plastoquinone (PQ) redox state in photosynthetic organisms. We demonstrated that light and/or heat treatment induced preferential photosystem (PS) I excitation by binding light-harvesting complex II (LHCII) proteins. The photosystem of wheat was in state 1 after dark overnight treatment, wherein PQ was oxidized and most of LHCII was not bound to PSI. At the onset of the light treatment [25 °C in the light (100 µmol photons m−2 s−1)], two major LHCIIs, Lhcb1 and Lhcb2 were phosphorylated, and the PSI–LHCII supercomplex formed within 5 min, which coincided with an increase in the PQ oxidation rate. Heat treatment at 40 °C of light-adapted wheat led to further LHCII protein phosphorylation of, resultant cyclic electron flow promotion, which was accompanied by ultrafast excitation of PSI and structural changes of thylakoid membranes, thereby protecting PSII from heat damage. These results suggest that LHCIIs are required for the functionality of wheat plant PSI, as it keeps PQ oxidized by regulating photochemical electron flow, thereby helping acclimation to environmental changes.
Keywords: Heat stress; Photosynthetic electron flow; Photosystem; Protein phosphorylation; State transition; Thylakoid membrane; Wheat

Differences in photosynthetic syndromes of four halophytic marsh grasses in Pakistan by Muhammad Moinuddin; Salman Gulzar; Abdul Hameed; Bilquees Gul; M. Ajmal Khan; Gerald E. Edwards (51-64).
Salt-tolerant grasses of warm sub-tropical ecosystems differ in their distribution patterns with respect to salinity and moisture regimes. Experiments were conducted on CO2 fixation and light harvesting processes of four halophytic C4 grasses grown under different levels of salinity (0, 200 and 400 mM NaCl) under ambient environmental conditions. Two species were from a high saline coastal marsh (Aeluropus lagopoides and Sporobolus tremulus) and two were from a moderate saline sub-coastal draw-down tidal marsh (Paspalum paspalodes and Paspalidium geminatum). Analyses of the carbon isotope ratios of leaf biomass in plants indicated that carbon assimilation was occurring by C4 photosynthesis in all species during growth under varying levels of salinity. In the coastal species, with increasing salinity, there was a parallel decrease in rates of CO2 fixation (A), transpiration (E) and stomatal conductance (g s), with no effect on water use efficiency (WUE). These species were adapted for photoprotection by an increase in the Mehler reaction with an increase in activity of PSII/CO2 fixed accompanied by high levels of antioxidant enzymes, superoxide dismutase and ascorbate peroxidase. The sub-coastal species P. paspalodes and P. geminatum had high levels of carotenoid pigments and non-photochemical quenching by the xanthophyll cycle.
Keywords: C4 photosynthesis; Carbon isotope discrimination; Halophytic grasses; Mehler reaction; Nonphotochemical quenching

A two-component nonphotochemical fluorescence quenching in eustigmatophyte algae by David Bína; Karel Bouda; Radek Litvín (65-77).
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; Nonphotochemical quenching; Xanthophyll cycle; Nannochloropsis ; Eustigmatophyceae

The reduction rate of photo-oxidized Photosystem I (PSI) with various natural and artificial electron donors have been well studied by transient absorption spectroscopy. The electron transfer rate from various donors to P700 + has been measured for a wide range of photosynthetic organisms encompassing cyanobacteria, algae, and plants. PSI can be a limiting component due to tedious extraction and purification methods required for this membrane protein. In this report, we have determined the in vivo, intracellular cytochrome c 6 (cyt c 6)/PSI ratio in Thermosynechococcus elongatus (T.e.) using quantitative Western blot analysis. This information permitted the determination of P700 + reduction kinetics via recombinant cyt c 6 in a physiologically relevant ratio (cyt c 6: PSI) with a Joliot-type, LED-driven, pump-probe spectrophotometer. Dilute PSI samples were tested under varying cyt c 6 concentration, temperature, pH, and ionic strength, each of which shows similar trends to the reported literature utilizing much higher PSI concentrations with laser-based spectrophotometer. Our results do however indicate kinetic differences between actinic light sources (laser vs. LED), and we have attempted to resolve these effects by varying our LED light intensity and duration. The standardized configuration of this spectrophotometer will also allow a more uniform kinetic analysis of samples in different laboratories. We can conclude that our findings from the LED-based system display an added total protein concentration effect due to multiple turnover events of P700 + reduction by cyt c 6 during the longer illumination regime.
Keywords: Photosystem I; Cytochrome c 6 ; Electron transfer; Flash photolysis; JTS-10; P700; Thermophilic cyanobacteria

Among marine phytoplankton groups, diatoms span the widest range of cell size, with resulting effects upon their nitrogen uptake, photosynthesis and growth responses to light. We grew two strains of marine centric diatoms differing by ~4 orders of magnitude in cell biovolume in high (enriched artificial seawater with ~500 µmol L−1 µmol L−1 NO3 ) and lower-nitrogen (enriched artificial seawater with <10 µmol L−1 NO3 ) media, across a range of growth light levels. Nitrogen and total protein per cell decreased with increasing growth light in both species when grown under the lower-nitrogen media. Cells growing under lower-nitrogen media increased their cellular allocation to RUBISCO and their rate of electron transport away from PSII, for the smaller diatom under low growth light and for the larger diatom across the range of growth lights. The smaller coastal diatom Thalassiosira pseudonana is able to exploit high nitrogen in growth media by up-regulating growth rate, but the same high-nitrogen growth media inhibits growth of the larger diatom species.
Keywords: Cell size; Chlorophyll; Phytoplankton; Nitrogen metabolism; RUBISCO; Thalassiosira pseudonana ; Thalassiosira punctigera

Ultrafast spectroscopy tracks carotenoid configurations in the orange and red carotenoid proteins from cyanobacteria by Václav Šlouf; Valentyna Kuznetsova; Marcel Fuciman; Céline Bourcier de Carbon; Adjélé Wilson; Diana Kirilovsky; Tomáš Polívka (105-117).
A quenching mechanism mediated by the orange carotenoid protein (OCP) is one of the ways cyanobacteria protect themselves against photooxidative stress. Here, we present a femtosecond spectroscopic study comparing OCP and RCP (red carotenoid protein) samples binding different carotenoids. We confirmed significant changes in carotenoid configuration upon OCP activation reported by Leverenz et al. (Science 348:1463–1466. doi: 10.1126/science.aaa7234 , 2015) by comparing the transient spectra of OCP and RCP. The most important marker of these changes was the magnitude of the transient signal associated with the carotenoid intramolecular charge-transfer (ICT) state. While OCP with canthaxanthin exhibited a weak ICT signal, it increased significantly for canthaxanthin bound to RCP. On the contrary, a strong ICT signal was recorded in OCP binding echinenone excited at the red edge of the absorption spectrum. Because the carbonyl oxygen responsible for the appearance of the ICT signal is located at the end rings of both carotenoids, the magnitude of the ICT signal can be used to estimate the torsion angles of the end rings. Application of two different excitation wavelengths to study OCP demonstrated that the OCP sample contains two spectroscopically distinct populations, none of which is corresponding to the photoactivated product of OCP.
Keywords: Orange carotenoid protein; Red carotenoid protein; Non-photochemical quenching; Intramolecular charge-transfer state; Ultrafast spectroscopy