Photosynthesis Research (v.116, #1)

Temporal and spectral characterization of the photosynthetic reaction center from Heliobacterium modesticaldum by Adrien Chauvet; Josephine Sarrou; Su Lin; Steven P. Romberger; John H. Golbeck; Sergei Savikhin; Kevin E. Redding (1-9).
A time-resolved spectroscopic study of the isolated photosynthetic reaction center (RC) from Heliobacterium modesticaldum reveals that thermal equilibration of light excitation among the antenna pigments followed by trapping of excitation and the formation of the charge-separated state P800 +A0 occurs within ~25 ps. This time scale is similar to that reported for plant and cyanobacterial photosystem I (PS I) complexes. Subsequent electron transfer from the primary electron acceptor A0 occurs with a lifetime of ~600 ps, suggesting that the RC of H. modesticaldum is functionally similar to that of Heliobacillus mobilis and Heliobacterium chlorum. The (A0  − A0) and (P800 + − P800) absorption difference spectra imply that an 81-OH-Chl a F molecule serves as the primary electron acceptor and occupies the position analogous to ec3 (A0) in PS I, while a monomeric BChl g pigment occupies the position analogous to ec2 (accessory Chl). The presence of an intense photobleaching band at 790 nm in the (A0  − A0) spectrum suggests that the excitonic coupling between the monomeric accessory BChl g and the 81-OH-Chl a F in the heliobacterial RC is significantly stronger than the excitonic coupling between the equivalent pigments in PS I.
Keywords: Heliobacterium modesticaldum ; Reaction center; Electrochromic shift; Accessory pigment; Bacteriochlorophyll g

Effect of chemical oxidation by ferricyanide on bacteriochlorophyll a (BChl a) in the Fenna–Matthews–Olson protein (FMO) was studied using absorbance and fluorescence spectroscopy at ambient and cryogenic temperatures. Partially selective oxidation of pigments bound to the antenna complex was achieved and the probable absorption wavelength corresponding to the recently discovered bacteriochlorophyll No. 8 of 806 nm was obtained by comparative analysis of the effect of chemical oxidation and the effect of different isolation procedures. Formation of a stable product identified as a chlorophyll a derivative occurred upon chemical oxidation. This new pigment remained bound within the pigment–protein complex, and exhibited an efficient energy transfer to BChl a. Furthermore, complex effects of the pigment oxidation upon the fluorescence yield of the FMO protein were observed. Utility of this approach based on chemical modifications for the investigation of the native regulatory mechanisms involved in the energy transfer in the FMO protein is discussed.
Keywords: FMO; Bacteriochlorophyll; Oxidation; Absorbance spectra; Fluorescence

Alleviation of heat damage to photosystem II by nitric oxide in tall fescue by Ke Chen; Liang Chen; Jibiao Fan; Jinmin Fu (21-31).
Nitric oxide (NO) has been found to mediate plant responses to heat stress. The objective of this study was to investigate the protective role of NO in the recovery process of photosystem II (PSII) in tall fescue (Festuca arundinacea) against heat stress. Treatment of tall fescue leaves with NO donor sodium nitroprusside significantly improved the overall behavior of PSII probed by the chlorophyll a fluorescence transients, while the inhibition of NO accumulation by 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (PTIO, a NO scavenger) plus N G-nitro-l-arginine-methyl ester (L-NAME, NO synthase inhibitor) dramatically disrupted the operation of PSII. Specifically, under heat stress, the exogenous NO reduced the initial fluorescence (F 0), increased the maximal quantum yield (F V/F M), and disappeared the K-step of 0.3 ms. By the analysis of the JIP-test, the exogenous NO improved the quantum yield of the electron transport flux from Q A to Q B (ET0/ABS), and decreased the trapped excitation flux per reaction center (RC) (TR0/RC), electron transport flux per RC (ET0/RC), and electron flux reducing end electron acceptors per RC (RE0/RC). In addition, the exogenous NO reduced the content of H2O2, O 2 •− , and malondialdehyde and electrolyte leakage of tall fescue leaves. These data suggest that exogenous NO could protect plants, increase the amount of activated RC and improve the electron transport from oxygen evolving complex to D1 protein. Moreover, quantitative RT-PCR revealed that, in the presence of hydrogen peroxide, NO induced the gene expression of psbA, psbB, and psbC, which encode proteins belonging to subunits of PSII core reaction center (Psb) complex. These findings indicate that, as an important strategy to protect plants against heat stress, NO could improve the recovery process of PSII by the up regulation of the transcriptions of genes encoding PSII core proteins.
Keywords: Nitric oxide; Photosystem II; OJIP transient; Tall fescue; Heat damage

2-epi-5-epi-Valiolone synthase activity is essential for maintaining phycobilisome composition in the cyanobacterium Anabaena variabilis ATCC 29413 when grown in the presence of a carbon source by Edward Spence; Samantha J. Bryan; Mohamed Lisfi; John Cullum; Walter C. Dunlap; J. Malcolm Shick; Conrad W. Mullineaux; Paul F. Long (33-43).
The cyclase 2-epi-5-epi-valiolone synthase (EVS) is reported to be a key enzyme for biosynthesis of the mycosporine-like amino acid shinorine in the cyanobacterium Anabaena variabilis ATCC 29413. Subsequently, we demonstrated that an in-frame complete deletion of the EVS gene had little effect on in vivo production of shinorine. Complete segregation of the EVS gene deletion mutant proved difficult and was achieved only when the mutant was grown in the dark and in a medium supplemented with fructose. The segregated mutant showed a striking colour change from native blue-green to pale yellow-green, corresponding to substantial loss of the photosynthetic pigment phycocyanin, as evinced by combinations of absorbance and emission spectra. Transcriptional analysis of the mutant grown in the presence of fructose under dark or light conditions revealed downregulation of the cpcA gene that encodes the alpha subunit of phycocyanin, whereas the gene encoding nblA, a protease chaperone essential for phycobilisome degradation, was not expressed. We propose that the substrate of EVS (sedoheptulose 7-phosphate) or possibly lack of its EVS-downstream products, represses transcription of cpcA to exert a hitherto unknown control over photosynthesis in this cyanobacterium. The significance of this finding is enhanced by phylogenetic analyses revealing horizontal gene transfer of the EVS gene of cyanobacteria to fungi and dinoflagellates. It is also conceivable that the EVS gene has been transferred from dinoflagellates, as evident in the host genome of symbiotic corals. A role of EVS in regulating sedoheptulose 7-phosphate concentrations in the photophysiology of coral symbiosis is yet to be determined.
Keywords: Fructose; Pentose phosphate pathway; Phycobilisome; Phycocyanin; Mycosporine-like amino acids; Sedoheptulose 7-phosphate

Photosynthetic electron flow changed considerably during desiccation and re-hydration of the intertidal macroalgae Porphyra haitanensis. Activities of both photosystem (PSI) and photosystem (PSII) increased significantly at moderate desiccation levels. Whereas PSII activity was abolished at an absolute water content (AWC) <24 %, PSI remained active with progressive decreases in AWC to values as low as 16 %. This result suggested that cyclic electron flow around PSI was still active after inactivation of linear electron flow following severe desiccation. Moreover, the PSI activity was restored more rapidly than that of PSII upon re-hydration. Pretreatment of the blades with 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU) suppressed PSII activity following desiccation to an AWC of ~16 % AWC. Cyclic electron flow around PSI decreased markedly in blades pretreated with DCMU than in blades without pretreatment of DCMU during re-hydration in seawater containing DCMU. All results suggested that the activity of PSII under desiccation conditions plays an important role in the operation of cyclic electron flow during desiccation and its recovery during re-hydration. Therefore, we proposed the PSII activity during desiccation could eventually lead to the accumulation of NADPH, which could serve as electron donor for P700+ and promote its recovery during re-hydration, thereby favoring the operation of cyclic electron flow.
Keywords: Cyclic electron flow; Desiccation; Photosystem I and II; Porphyra haitanensis

The stoichiometry and energetics of oxygenic phototrophic growth by Igor G. Minkevich; Polina V. Fursova; Lada D. Tjorlova; Anatoly A. Tsygankov; Galina Yu. Riznichenko (55-78).
The values of gross metabolic flows in cells are essentially interconnected due to conservation laws of chemical elements and interrelations of biochemical coupling. Therefore, the overall stoichiometry of cellular metabolism, such as the biomass quantum yield, the ratio between linear and circular flows via the electron transport chain, etc., can be calculated using balances of metabolic flows in the network branching points and coupling ratios related to ATP formation and expenditures. This work has studied the energetic stoichiometry of photosynthetic cells by considering the transfer of reductivity in the course of biochemical reactions. This approach yielded rigorous mathematical expressions for biomass quantum yield and other integral bioenergetic indices of cellular growth as functions of ATP balance parameters. The effect of cellular substance turnover has been taken into account. The obtained theoretical estimation of biomass quantum yield is rather close to experimental data which confirms the predictive capacity of this approach.
Keywords: Photosystems; Electron transport chain; High-energy bonds; Reductivity; Cell bioenergetics; Biomass quantum yield

Regulation of photosynthesis during heterocyst differentiation in Anabaena sp. strain PCC 7120 investigated in vivo at single-cell level by chlorophyll fluorescence kinetic microscopy by Naila Ferimazova; Kristina Felcmanová; Eva Šetlíková; Hendrik Küpper; Iris Maldener; Günther Hauska; Barbora Šedivá; Ondřej Prášil (79-91).
Changes of photosynthetic activity in vivo of individual heterocysts and vegetative cells in the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120 during the course of diazotrophic acclimation were determined using fluorescence kinetic microscopy (FKM). Distinct phases of stress and acclimation following nitrogen step-down were observed. The first was a period of perception, in which the cells used their internally stored nitrogen without detectable loss of PS II activity or pigments. In the second, the stress phase of nitrogen limitation, the cell differentiation occurred and an abrupt decline of fluorescence yield was observed. This decline in fluorescence was not paralleled by a corresponding decline in photosynthetic pigment content and PS II activity. Both maximal quantum yield and sustained electron flow were not altered in vegetative cells, only in the forming heterocysts. The third, acclimation phase started first in the differentiating heterocysts with a recovery of PS II photochemical yields $$F_{ ext{v}} /F_{ ext{m}} ,;F^{prime}_{ ext{v}} /F^{prime}_{ ext{m}}.$$ F v / F m , F v ′ / F m ′ . Afterwards, the onset of nitrogenase activity was observed, followed by the restoration of antenna pigments in the vegetative cells, but not in the heterocysts. Surprisingly, mature heterocysts were found to have an intact PS II as judged by photochemical yields, but a strongly reduced PS II-associated antenna as judged by decreased F 0. The possible importance of the functional PS II in heterocysts is discussed. Also, the FKM approach allowed to follow in vivo and evaluate the heterogeneity in photosynthetic performance among individual vegetative cells as well as heterocysts in the course of diazotrophic acclimation. Some cells along the filament (so-called “superbright cells”) were observed to display transiently increased fluorescence yield, which apparently proceeded by apoptosis.
Keywords: Acclimation to abiotic stress; Heterocyst differentiation; Nitrogen fixation; Two-dimensional (imaging) measurements of chlorophyll fluorescence kinetics of individual cells; Topography of photosynthetic performance at single-cell level

Characterization of photosystem I in rice (Oryza sativa L.) seedlings upon exposure to random positioning machine by Boya Chen; Aihong Zhang; Qingtao Lu; Tingyun Kuang; Congming Lu; Xiaogang Wen (93-105).
To gain a better understanding of how photosynthesis is adapted under altered gravity forces, photosynthetic apparatus and its functioning were investigated in rice (Oryza sativa L.) seedlings grown in a random positioning machine (RPM). A decrease in fresh weight and dry weight was observed in rice seedlings grown under RPM condition. No significant changes were found in the chloroplast ultrastructure and total chlorophyll content between the RPM and control samples. Analyses of chlorophyll fluorescence and thermoluminescence demonstrate that PSII activity was unchanged under RPM condition. However, PSI activity decreased significantly under RPM condition. 77 K fluorescence emission spectra show a blue-shift and reduction of PSI fluorescence emission peak in the RPM seedlings. In addition, RPM caused a significant decrease in the amplitude of absorbance changes of P700 at 820 nm (A 820) induced by saturated far-red light. Moreover, the PSI efficiency (Φ I) decreased significantly under RPM condition. Immunoblot and blue native gel analyses further illustrate that accumulation of PSI proteins was greatly decreased in the RPM seedlings. Our results suggest that PSI, but not PSII, is down-regulated under RPM condition.
Keywords: Random positioning machine; Clinorotation; Rice (Oryza sativa L.); Photosynthesis; Photosystem I; Photosystem II