Ssl3451 is Important for Accumulation of NDH-1 Assembly Intermediates in the Cytoplasm of Synechocystis sp. Strain PCC 6803.

Two mutants sensitive to high light for growth and impaired in NDH-1 activity were isolated from a transposon-tagged library of Synechocystis sp. strain PCC 6803. Both mutants were tagged in the ssl3451 gene encoding a hypothetical protein, which shares a significant homology with the Arabidopsis (Arabidopsis thaliana) CHLORORESPIRATORY REDUCTION 42 (CRR42). In Arabidopsis, CRR42 associates only with an NDH-1 hydrophilic arm assembly intermediate (NAI) of about 400 kDa (NAI400), one of total three NAIs (NAI800, NAI500 and NAI400), and its deletion has little, if any, effect on accumulation of any NAIs in the stroma. In comparison, the ssl3451 product was localized mainly in the cytoplasm and associates with two NAIs of about 300 kDa (NAI300) and 130 kDa (NAI130). Deletion of Ssl3451 reduced the abundance of the NAI300 complex to levels no longer visible on gels and of the NAI130 complex to a low level, thereby impeding the assembly process of NDH-1 hydrophilic arm. Further, Ssl3451 interacts with another assembly factor Ssl3829 and they have a similar effect on accumulation of NAIs and NdhI maturation factor Slr1097 in the cytoplasm. We thus propose that Ssl3451 plays an important role in accumulation of the NAI300 and NAI130 complexes in the cytoplasm via its interacting protein Ssl3829.

Light Intensity is Important for Hydrogen Production in NaHSO3-Treated Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii is a unicellular green alga that can use light energy to produce H2 from H2O in the background of NaHSO3 treatment. However, the role of light intensity in such H2 production remains elusive. Here, light intensity significantly affected the yield of H2 production in NaHSO3-treated C. reinhardtii, which was consistent with its effects on the content of O2 and the expression and activity of hydrogenase. Further, NaHSO3 was found to be able to remove O2 via a reaction of bisulfite with superoxide anion produced at the acceptor side of PSI, and light intensity affected the reaction rate significantly. Accordingly, high light and strong light but not low light can create an anaerobic environment, which is important to activate hydrogenase and produce H2. Based on the above results, we conclude that light intensity plays an important role in removing O2 and consequently activating hydrogenase and producing H2 in NaHSO3-treated C. reinhardtii.

NdhV Is a Subunit of NADPH Dehydrogenase Essential for Cyclic Electron Transport in Synechocystis sp. Strain PCC 6803.

Two mutants sensitive to heat stress for growth and impaired in NADPH dehydrogenase (NDH-1)-dependent cyclic electron transport around photosystem I (NDH-CET) were isolated from the cyanobacterium Synechocystis sp. strain PCC 6803 transformed with a transposon-bearing library. Both mutants had a tag in the same sll0272 gene, encoding a protein highly homologous to NdhV identified in Arabidopsis (Arabidopsis thaliana). Deletion of the sll0272 gene (ndhV) did not influence the assembly of NDH-1 complexes and the activities of CO2 uptake and respiration but reduced the activity of NDH-CET. NdhV interacted with NdhS, a ferredoxin-binding subunit of cyanobacterial NDH-1 complex. Deletion of NdhS completely abolished NdhV, but deletion of NdhV had no effect on the amount of NdhS. Reduction of NDH-CET activity was more significant in ΔndhS than in ΔndhV. We therefore propose that NdhV cooperates with NdhS to accept electrons from reduced ferredoxin.

Identification of a c-type heme oxygenase and its function during acclimation of cyanobacteria to nitrogen fluctuations.

The mechanisms of acclimating to a nitrogen-fluctuating environment are necessary for the survival of aquatic cyanobacteria in their natural habitats, but our understanding is still far from complete. Here, the synthesis of phycobiliprotein is confirmed to be much earlier than that of photosystem components during recovery from nitrogen chlorosis and an unknown protein Ssr1698 is discovered to be involved in this synthetic process. The unknown protein is further identified as a c-type heme oxygenase (cHO) in tetrapyrrole biosynthetic pathway and catalyzes the opening of heme ring to form biliverdin IXα, which is required for phycobilin production and ensuing phycobiliprotein synthesis. In addition, the cHO-dependent phycobiliprotein is found to be vital for the growth of cyanobacterial cells during chlorosis and regreening through its nitrogen-storage and light-harvesting functions, respectively. Collectively, the cHO expressed preferentially during recovery from nitrogen chlorosis is identified in photosynthetic organisms and the dual function of this enzyme-dependent phycobiliprotein is proposed to be an important mechanism for acclimation of aquatic cyanobacteria to a nitrogen-fluctuating environment.

Identification of novel Ssl0352 protein (NdhS), essential for efficient operation of cyclic electron transport around photosystem I, in NADPH:plastoquinone oxidoreductase (NDH-1) complexes of Synechocystis sp. PCC 6803.

Cyanobacterial NADPH:plastoquinone oxidoreductase, or type I NAD(P)H dehydrogenase, or the NDH-1 complex is involved in plastoquinone reduction and cyclic electron transfer (CET) around photosystem I. CET, in turn, produces extra ATP for cell metabolism particularly under stressful conditions. Despite significant achievements in the study of cyanobacterial NDH-1 complexes during the past few years, the entire subunit composition still remains elusive. To identify missing subunits, we screened a transposon-tagged library of Synechocystis 6803 cells grown under high light. Two NDH-1-mediated CET (NDH-CET)-defective mutants were tagged in the same ssl0352 gene encoding a short unknown protein. To clarify the function of Ssl0352, the ssl0352 deletion mutant and another mutant with Ssl0352 fused to yellow fluorescent protein (YFP) and the His(6) tag were constructed. Immunoblotting, mass spectrometry, and confocal microscopy analyses revealed that the Ssl0352 protein resides in the thylakoid membrane and associates with the NDH-1L and NDH-1M complexes. We conclude that Ssl0352 is a novel subunit of cyanobacterial NDH-1 complexes and designate it NdhS. Deletion of the ssl0352 gene considerably impaired the NDH-CET activity and also retarded cell growth under high light conditions, indicating that NdhS is essential for efficient operation of NDH-CET. However, the assembly of the NDH-1L and NDH-1M complexes and their content in the cells were not affected in the mutant. NdhS contains a Src homology 3-like domain and might be involved in interaction of the NDH-1 complex with an electron donor.

Genome-Wide Insights Into the Organelle Translocation of Photosynthetic NDH-1 Genes During Evolution.

Translocation of chloroplast-located genes to mitochondria or nucleus is considered to be a safety strategy that impedes mutation of photosynthetic genes and maintains their household function during evolution. The organelle translocation strategy is also developed in photosynthetic NDH-1 (pNDH-1) genes but its understanding is still far from complete. Here, we found that the mutation rate of the conserved pNDH-1 genes was gradually reduced but their selection pressure was maintained at a high level during evolution from cyanobacteria to angiosperm. By contrast, oxygenic photosynthesis-specific (OPS) pNDH-1 genes had an opposite trend, explaining the reason why they were transferred from the reactive oxygen species (ROS)-enriched chloroplast to the ROS-barren nucleus. Further, genome-wide sequence analysis supported the possibility that all conserved pNDH-1 genes lost in chloroplast genomes of Chlorophyceae and Pinaceae were transferred to the ROS-less mitochondrial genome as deduced from their truncated pNDH-1 gene fragments. Collectively, we propose that the organelle translocation strategy of pNDH-1 genes during evolution is necessary to maintain the function of the pNDH-1 complex as an important antioxidant mechanism for efficient photosynthesis.

New insights into the effect of NdhO levels on cyanobacterial cell death triggered by high temperature.

NdhO, a regulatory oxygenic photosynthesis-specific subunit, is close to the ferredoxin-binding site of cyanobacterial NDH-1, and its levels are negatively associated with the rates of cyclic electron transfer around PSI mediated by NDH-1 (NDH-CET). However, the effect of NdhO levels on cyanobacterial cell death triggered by high temperature remains elusive. Here, our results uncovered a synergistic effect of NdhO levels on the cell death and reactive oxygen species (ROS) accumulation when cyanobacterial cells grown at 30°C for 1 day were transferred to 45°C for 2 days. Such synergistic effect was found to be closely associated with the activities of NDH-CET and CO2 assimilation during high temperature. Collectively, we propose that the effect of NdhO levels on the cyanobacterial cell bleaching and cell death triggered by high temperature is a result of influencing production of ROS by NDH-CET, which is considered to be vital to balance the ATP/NADPH ratio and improve the Calvin-Benson cycle.

Study on a new "One-stop-shop" scan protocol combining brain CT perfusion and head-and-neck CT angiography by using 256-detector CT for stroke patients.

PURPOSE: We sought to evaluate the performance of a new "one-stop-shop" scan protocol combining brain computed tomography perfusion (CTP) and head-and-neck CT angiography (CTA) imaging for acute stroke patients using a 256-detector CT scanner. METHOD: From March to August 2020, 60 patients (30 men and 30 women) aged 22-88 years with suspected acute stroke were enrolled and randomly divided into 2 groups to undergo brain CTP and head-and-neck CTA with a 256-detector CT system. Group A used traditional scan protocol with a separate brain CTP and head-and-neck CT examination that included non-contrast-enhanced and contrast-enhanced acquisitions; group B used the new "one-stop-shop" scan protocol with head-and-neck CTA data inserted into brain CTP scans at the peak time (PT) of the arterial phase. The insertion point of the head-and-neck CTA data was determined by a test bolus. The examination time, contrast dose, radiation dose, and image quality were compared between the groups. RESULTS: The total contrast dose was reduced by 40% in group B compared to group A (60 mL vs. 100 mL). The imaging time was 52.5 ± 2.6 s in group B and 74.9 ± 3.3 s in group A, showing a reduction of approximately 43% in group B. There was no significant difference in image quality both quantitatively and qualitatively between the groups (all P > 0.05). Group B had a slight reduction in dose length product (1139.0 ± 45.3 vs. 1211.6 ± 31.9 mGy·cm, P < 0.001). CONCLUSIONS: The proposed "one-stop-shop" scan protocol combining brain CTP and head-and-neck CTA on a 256-detector CT system can reduce imaging time and contrast dose, without affecting image quality or perfusion results, compared to the traditional protocol of separating the examinations.

Mechanistic insights into pH-dependent H2 photoproduction in bisulfite-treated Chlamydomonas cells.

BACKGROUND: Bisulfite addition is an important H2 photoproduction strategy that removes O2 and activates hydrogenase. The pH values of cell cultures can change the ratio of bisulfite to sulfite, which may affect H2 photoproduction. However, little is known regarding the pH effect of bisulfite addition on H2 photoproduction and relevant underlying mechanism. RESULTS: Here, changes in H2 photoproduction with different initial extracellular pH values showed a parabolic distribution and a pH of 8.0 is an optimal value for H2 photoproduction in Chlamydomonas reinhardtii cells treated with bisulfite. Compared to the growth pH (pH 7.3), increased photoproduction of H2 at this optimal pH was primarily caused by a relatively high residual activity of photosystem II (PSII), which provides a relatively plentiful source of electrons for H2 photoproduction. Such increased H2 photoproduction was most likely a result of decreased the ratio of bisulfite to sulfite, consistent with the result that the toxicity of bisulfite on PSII was much more than that of sulfite. This possibility was corroborated by the result that treatment with a combination of 7 mM bisulfite and 6 mM sulfite further enhanced H2 photoproduction compared with 13 mM bisulfite alone. CONCLUSIONS: Collectively, our findings provide novel mechanistic insights into pH-dependent H2 photoproduction in C. reinhardtii cells treated with bisulfite, and demonstrate that sulfite addition is another important strategy for H2 photoproduction, just like bisulfite addition.

The response of electron transport mediated by active NADPH dehydrogenase complexes to heat stress in the cyanobacterium Synechocystis 6803.

The electron-transport machinery in photosynthetic membranes is known to be very sensitive to heat. In this study, the rate of electron transport (ETR) driven by photosystem I (PSI) and photosystem II (PSII) during heat stress in the wild-type Synechocystis sp. strain PCC 6803 (WT) and its ndh gene inactivation mutants DeltandhB (M55) and DeltandhD1/ndhD2 (D1/D2) was simultaneously assessed by using the novel Dual-PAM-100 measuring system. The rate of electron transport driven by the photosystems (ETR(PSs)) in the WT, M55, and D1/D2 cells incubated at 30 degrees C and at 55 degrees C for 10 min was compared. Incubation at 55 degrees C for 10 min significantly inhibited PSII-driven ETR (ETR(PSII)) in the WT, M55 and D1/D2 cells, and the extent of inhibition in both the M55 and D1/D2 cells was greater than that in the WT cells. Further, PSI-driven ETR (ETR(PSI)) was stimulated in both the WT and D1/D2 cells, and this rate was increased to a greater extent in the D1/D2 than in the WT cells. However, ETR(PSI) was considerably inhibited in the M55 cells. Analysis of the effect of heat stress on ETR(PSs) with regard to the alterations in the 2 active NDH-1 complexes in the WT, M55, and D1/D2 cells indicated that the active NDH-1 supercomplex and mediumcomplex are essential for alleviating the heat-induced inhibition of ETR(PSII) and for accelerating the heat-induced stimulation of ETR(PSI), respectively. Further, it is believed that these effects are most likely brought about by the electron transport mediated by each of these 2 active NDH-1 complexes.

A Stepwise NaHSO3 Addition Mode Greatly Improves H2 Photoproduction in Chlamydomonas reinhardtii.

NaHSO3 addition greatly increases the yield of H2 photoproduction in a unicellular green alga Chlamydomonas reinhardtii through removing O2 and activating hydrogenase but significantly impairs the activity of PSII, an electron source for H2 photoproduction. Here, a stepwise addition mode of total 13 mM NaHSO3, an optimal concentration for H2 photoproduction of C. reinhardtii identified in a previous one step addition method, significantly improved H2 photoproduction. Such improvement was believed to be the result of increased residual PSII activity in an anaerobic background, but was at least independent of two alternative electron sinks for H2 photoproduction, cyclic electron transport around PSI and CO2 assimilation. Based on the above results, we propose that increased residual PSII activity in an anaerobic environment is an efficient strategy to enhance H2 photoproduction in C. reinhardtii, and the stepwise NaHSO3 addition mode is a case study in the strategy.

A simple approach for the efficient production of hydrogen from Taihu Lake Microcystis spp. blooms.

The death and subsequent decomposition of algal blooms is capable of depleting dissolved O2 to anaerobic levels, and this can de-inactivate hydrogenases. Inspired by this fact, a simple method for efficient H2 production from algal bloom biomass was developed. Direct transfer of Taihu Lake Microcystis spp. blooms into dark conditions resulted in H2 evolution, and yield was much greater than compared to Microcystis spp. cultured in the laboratory and reported previously in the literature. Further, efficient H2 production was inhibited significantly by light, which was most likely due to reduced O2 content and the stimulation of hydrogenase activity. Therefore, a simple approach for efficient H2 production from Taihu Lake Microcystis spp. blooms is presented. Furthermore, a post-treatment strategy for dealing with large quantities of refloated algal blooms is proposed.

Treatment with NaHSO3 greatly enhances photobiological H2 production in the green alga Chlamydomonas reinhardtii.

Treatment with NaHSO3 induces a 10-fold increase in H2 photoproduction in the filamentous N2-fixing cyanobacterium Anabaena sp. strain PCC 7120. However, it is unclear whether this treatment also increases H2 photoproduction in green alga. In this study, treatment with 13 mM NaHSO3 resulted in about a 200-fold increase in H2 production in Chlamydomonas reinhardtii, and this increase was most probably the result of reduced O2 content and enhanced hydrogenase activity. Compared to the conventional strategy of sulfur deprivation, NaHSO3 treatment results in a higher maximum rate of H2 photoproduction, greater efficiency of conversion of light energy into H2, shorter half-time to produce the maximum accumulated H2 levels, and reduced costs because no centrifugation is involved. We therefore conclude that NaHSO3 treatment is an efficient, rapid, and economic strategy for improving photobiological H2 production in the green alga C. reinhardtii.