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Cyanobacteria are a group of oxygenic photosynthetic bacteria with great potentials in biotechnological applications and advantages as models for photosynthesis research. The subcellular localizations of the majority of proteins in any cyanobacteria remain undetermined, representing a major challenge in using cyanobacteria for both basic and industrial researches. Here, using label-free quantitative proteomics, we map 2027 proteins of Synechocystis sp. PCC6803, a model cyanobacterium, to different subcellular compartments and generate a proteome atlas with such information. The atlas leads to numerous unexpected but important findings, including the predominant localization of the histidine kinases Hik33 and Hik27 on the thylakoid but not the plasma membrane. Such information completely changes the concept regarding how the two kinases are activated. Together, the atlas provides subcellular localization information for nearly 60% proteome of a model cyanobacterium, and will serve as an important resource for the cyanobacterial research community.

Sheepgrass [Leymus chinensis (Trin.) Tzvel] is a valuble forage plant highly significant to regional grassland productivity of Euro-Asia steppes. Although effects of environmental stress including drought have been studied, impact of nutrient deficiency in particular phosphate (Pi), one of the essential macronutrient, is not reported. Here, we investigated low-Pi effect on its photosynthetic apparatus via physiological and biochemical analysis. We show that PSII activity was significantly reduced based on chlorophyll fluorescence measurements. We observed decreased amount of core proteins of PSII by immunoblot analysis. Further analysis of thylakoid membranes using 2D-BN/SDS-PAGE and immunoblot detection demonstrated that the amount of PSII complexes was closely correlated with the Pi levels within the range. Together with reduced number of thylakoid grana stackings observed, we suggest that the maintenance of PSII is impaired under Pi-limited condition. Moreover, enzyme activity assays revealed that the activity of several ROS scavenger enzymes was stimulated by low-Pi treatment. Based on these experimental results we conclude that PSII is the component of photosynthetic apparatus most sensitive to Pi supply and the enhanced anti-ROS activity is mainly subjective to protection of PSII against low Pi-induced photo-oxidative stress in the organism.

Global climate warming has a crucial impact on many terrestrial ecosystems, including temperate steppe. In addition, phosphate deficiency is known to be the common deficiency in soils worldwide due to the low availability of the phosphate nutrient in the form of inorganic phosphate anions (Pi). Consequently, in the future, land plants are likely to simultaneously encounter heat stress and phosphate deficiency more frequently. Sheepgrass 〔Leymus chinensis (Trin.) Tzvel〕is a dominant perennial forage plant highly significant to grass productivity of Eurasian temperate grasslands. Though effects of environmental stress including drought and Pi starvation have been studied, the combined eff ;ects of phosphate deficiency and heat stress on plant physiology remain largely unclear. Here, we investigated the combined eff ;ects of heat stress and phosphate deficiency on above-ground tissue growth and photochemical properties of L. chinensis using in vivo chlorophyll fluorescence spectroscopy. We observed remarkable phenotypic alterations of reduced shoot growth and considerable leaf chlorosis in L. chinensis seedlings under the combined stress condition. Also, we compared changes in photochemical activity between the control and the corresponding stressed seedlings. Based on chlorophyll fluorescence analysis, impairment of PSI was more severe than that of PSII in the seedlings treated with the combined stress. Compared to the control, PSI and PSII activity decreased up to 35.5% and 30%, respectively, under the combined-stress condition. Moreover, our data show that the decreased photosynthetic activity is not the sum of the single-stressed conditions. These results combined with the distinction of other photochemical parameters indicate that a complex interaction between Pi-deficiency and heat stress may exist in the forage plant.

Glutaminase is widely distributed among microorganisms and mammals with important functions. Little is known regarding the biochemical properties and functions of the deamidating enzyme glutaminase in cyanobacteria. In this study a putative glutaminase encoded by gene slr2079 in Synechocystis sp. PCC 6803 was investigated. The slr2079 was expressed as histidine-tagged fusion protein in Escherichia coli. The purified protein possessed glutaminase activity, validating the functional assignment of the genomic annotation. The apparent K (m) value of the recombinant protein for glutamine was 26.6 +/- 0.9 mmol/L, which was comparable to that for some of other microbial glutaminases. Analysis of the purified protein revealed a two-fold increase in catalytic activity in the presence of 1 mol/L Na(+). Moreover, the K (m) value was decreased to 12.2 +/- 1.9 mmol/L in the presence of Na(+). These data demonstrate that the recombinant protein Slr2079 is a glutaminase which is regulated by Na(+) through increasing its affinity for substrate glutamine. The slr2079 gene was successfully disrupted in Synechocystis by targeted mutagenesis and the Deltaslr2079 mutant strain was analyzed. No differences in cell growth and oxygen evolution rate were observed between Deltaslr2079 and the wild type under standard growth conditions, demonstrating slr2079 is not essential in Synechocystis. Under high salt stress condition, however, Deltaslr2079 cells grew 1.25-fold faster than wild-type cells. Moreover, the photosynthetic oxygen evolution rate of Deltaslr2079 cells was higher than that of the wild-type. To further characterize this phenotype, a number of salt stress-related genes were analyzed by semi-quantitative RT-PCR. Expression of gdhB and prc was enhanced and expression of desD and guaA was repressed in Deltaslr2079 compared to the wild type. In addition, expression of two key enzymes of ammonium assimilation in cyanobacteria, glutamine synthetase (GS) and glutamate synthase (GOGAT) was examined by semi-quantitative RT-PCR. Expression of GOGAT was enhanced in Deltaslr2079 compared to the wild type while GS expression was unchanged. The results indicate that slr2079 functions in the salt stress response by regulating the expression of salt stress related genes and might not play a major role in glutamine breakdown in Synechocystis.

Cyanobacteria are ancient photosynthetic prokaryotes that have adapted successfully to adverse environments including high-light irradiation. Although it is known that the repair of photodamaged photosystem II (PSII) in the organisms is a highly regulated process, our knowledge of the molecular components that regulate each step of the process is limited. We have previously identified a hypothetical protein Slr0151 in the membrane fractions of cyanobacterium Synechocystis sp. PCC 6803. Here, we report that Slr0151 is involved in PSII repair of the organism. We generated a mutant strain (Δslr0151) lacking the protein Slr0151 and analyzed its characteristics under normal and high-light conditions. Targeted deletion of slr0151 resulted in decreased PSII activity in Synechocystis. Moreover, the mutant exhibited increased photoinhibition due to impairment of PSII repair under high-light condition. Further analysis using in vivo radioactive labeling and 2-D blue native/sodium dodecylsulfate polyacrylamide gel electrophoresis indicated that the PSII repair cycle was hindered at the levels of D1 synthesis and disassembly and/or assembly of PSII in the mutant. Protein interaction assays demonstrated that Slr0151 interacts with D1 and CP43 proteins. Taken together, these results indicate that Slr0151 plays an important role in regulating PSII repair in the organism under high-light stress condition.

The genome of unicellular green alga Chlamydomonas reinhardtii contains four genes encoding B-type methionine sulfoxide reductases, MSRB1.1, MSRB1.2, MSRB2.1, and MSRB2.2, with functions largely unknown. To understand the cell defense system mediated by the methionine sulfoxide reductases in Chlamydomonas, we analyzed expression and physiological roles of the MSRBs under different abiotic stress conditions using immunoblotting and quantitative polymerase chain reaction (PCR) analyses. We showed that the MSRB2.2 protein was accumulated in cells treated with high light (1,300 µE/m² per s), whereas MSRB1.1 was accumulated in the cells under 1 mmol/L H₂O₂ treatment or sulfur depletion. We observed that the cells with the MSRB2.2 knockdown and overexpression displayed increased and decreased sensitivity to high light, respectively, based on in situ chlorophyll a fluorescence measures. We also observed that the cells with the MSRB1.1 knockdown and overexpression displayed decreased and increased tolerance to sulfur-depletion and oxidative stresses, respectively, based on growth and H₂-producing performance. The physiological implications revealed from the experimental data highlight the importance of MSRB2.2 and MSRB1.1 in protecting Chlamydomonas cells against adverse conditions such as high-light, sulfur-depletion, and oxidative stresses.