Effect of growth temperature on the composition of the photosynthetic apparatus in a thermophilic cyanobacterium (Synechococcus sp.)

Abstract A thermophilic strain of Synechococcus sp. was grown under light-limited conditions at its optimum temperature of 58°C and at 38°C in order to investigate the effect of growth temperature on the composition of the photosynthetic apparatus. Cells grown at 38°C had a ratio of Photosystem I to Photosystem II of 2.2, close to that known to occur in unstressed mesophilic strains of Synechococcus . Growth at the higher temperature led to a doubling of the number of Photosystem I reaction centres per cell whilst Photosystem II remained essentially unchanged, causing the Photosystem I/Photosystem II ratio to rise to 4.1. These changes were correlated with an increase in the number of concentric layers of thylakoid membranes from four to nine. It is suggested that these changes result from a higher relative demand for energy (ATP) during growth at elevated temperatures.     

Purification of RuBisCO from the thermophilic cyanobacterium Synechococcus sp. strain a-1

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the key enzyme of the Calvin Benson cycle, has been purified from a thermophilic cyanobacterium, Synechococcus sp. strain a-1 and characterized. The enzyme is an L₈S₈-type hexadecamer with a molecular mass of 530 kDa. The enzyme was stable against heat treatment up to 70°C, which is the highest value among the RuBisCOs so far purified. The K_m value for ribulose bisphosphate on the carboxylase activity was substantially higher than those observed for RuBisCOs obtained from mesophilic autotrophs. The N-terminal amino acid sequence for the large subunit of the enzyme was highly similar to those of the other cyanobacteria despite the significant differences in heat stability.     

Acclimation of Arabidopsis thaliana to the light environment: Changes in composition of the photosynthetic apparatus

Acclimation to changes in the light environment was investigated in Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta. Plants grown under four light regimes showed differences in their development, morphology, photosynthetic performance and in the composition of the photosynthetic apparatus. Plants grown under high light showed higher maximum rates of oxygen evolution and lower levels of light-harvesting complexes than their low light-grown counterparts; plants transferred to low light showed rapid changes in maximum photosynthetic rate and chlorophyll-a/b ratio as they became acclimated to the new environment. In contrast, plants grown under lights of differing spectral quality showed significant differences in the ratio of photosystem II to photosystem I. These changes are consistent with a model in which photosynthetic metabolism provides signals which regulate the composition of the thylakoid membrane.Abbreviations Aac1 gene encoding actin - Chl chlorophyll - F far-red-enriched light (R:FR = 0.72) - FR far-red light - H high light (400 mol · m^–2 · s^–1) - L low light (100 ml · m^–2 · s^–1) - LHCII light-harvesting complex of PSII - Lhcb genes encoding the proteins of LHCII - R red light - Rbcs genes encoding the small subunit of Rubisco - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - W white light (R:FR = 1.40) This work was supported by Natural Environment Research Council Grant No. GR3/7571A. We would like to thank H. Smith (Botany Department, University of Leicester) and E. Murchie (University of Sheffield) for helpful discussions.     

Oxygen and light effects on the expression of the photosynthetic apparatus in Bradyrhizobium sp. C7T1 strain

Photosynthetic bradyrhizobia are nitrogen-fixing symbionts colonizing the stem and roots of some leguminous plants like Aeschynomene. The effect of oxygen and light on the formation of the photosynthetic apparatus of Bradyrhizobium sp. C7T1 strain is described here. Oxygen is required for growth, but at high concentration inhibits the synthesis of bacteriochlorophyll (BChl) and of the photosynthetic apparatus. However, we show that in vitro, aerobic photosynthetic electron transport occurred leading to ADP photophosphorylation. The expression of the photosynthetic apparatus was regulated by oxygen in a manner which did not agree with earlier results in other photosynthetic bradyrhizobia since BChl accumulation was the highest under microaerobic conditions. This strain produces photosynthetic pigments when grown under cyclic illumination or darkness. However, under continuous white light illumination, a Northern blot analysis of the puf operon showed that, the expression of the photosynthetic genes of the antenna was considerable. Under latter conditions BChl accumulation in the cells was dependent on the oxygen concentration. It was not detectable at high oxygen tensions but became accumulated under low oxygen (microaerobiosis). It is known that in photosynthetic bradyrhizobia bacteriophytochrome photoreceptor (BphP) partially controls the synthesis of the photosystem in response to light. In C7T1 strain far-red light illumination did not stimulate the synthesis of the photosynthetic apparatus suggesting the presence of a non-functional BphP-mediated light regulatory mechanism.     

Calcium carbonate formation by Synechococcus sp. strain PCC 8806 and Synechococcus sp. strain PCC 8807

Precipitation of CaCO3 catalyzed by the growth and physiology of cyanobacteria in the genus Synechococcus represents a potential mechanism for sequestration of atmospheric CO2 produced during the burning of coal for power generation. Synechococcus sp. strain PCC 8806 and Synechococcus sp. strain PCC 8807 were tested in microcosm experiments for their ability to calcify when exposed to a fixed calcium concentration of 3.4 mM and dissolved inorganic carbon concentrations of 0.5, 1.25 and 2.5 mM. Synechococcus sp. strain PCC 8806 removed calcium continuously over the duration of the experiment producing approximately 18.6 mg of solid phase calcium. Calcium removal occurred over a two-day time period when Synechococcus sp. strain PCC 8807 was tested and only 8.9 mg of solid phase calcium was produced. Creation of an alkaline growth environment catalyzed by the physiology of the cyanobacteria appeared to be the primary factor responsible for CaCO3 precipitation in these experiments.     

Changes in the photosynthetic apparatus of diatoms in response to low and high light intensities

The centric diatom Cyclotella cryptica and two strains of the pennate diatom Phaeodactylum tricornutum were grown under low and high light intensities (300 lux and 3,000 lux) over 4-6 weeks. Growth was monitored by repetitive cell count. The culture media were replaced weekly to avoid morphological and biochemical alterations caused by nutrient depletion. The ultrastructure of the cells was examined by transmission electron microscopy. Alterations in the light-harvesting antenna systems were investigated by Western immunoblotting. Both diatoms reduced the plastid area, i.e. decreased the amount of thylakoid lamellae, under high light intensity. The thylakoids still ran in groups of three with parallel orientation within the chloroplasts. The girdle band lamellae were not affected. The amounts of storage compounds and vacuoles increased. SDS-PAGE of total cell protein followed by Western immunoblotting with antisera directed against subunits of the light-harvesting antenna systems of C. cryptica (cc-antiserum) and the cryptophyte Cryptomonas maculata (cmac-antiserum) revealed that both diatoms reduced the amount of antenna polypeptides under increased light intensity. The cc-antiserum immunodecorated two bands with relative molecular masses (Mr) of 18,000 and 22,000 in C. cryptica. Both decreased under high light conditions to 67.2 +/- 6.1%. Five to seven bands in the Mr range of 14,000-27,000 were recognized in P. tricornutum. They decreased to 83 +/- 5.3%. Furthermore, the immunolabeling pattern for both strains differed under the two light regimes. The cmac-antiserum immunodecorated two polypeptides with Mr of 24,000 and 23,000 in C. cryptica, while both strains of P. tricornutum had five polypeptides in the Mr range of 14,000-24,000 that showed some differences in staining intensities between the two strains and in response to the light intensity applied.     

Multiple forms of chlorophyll-protein complexes from a thermophilic cyanobacterium Synechococcus sp

Eight chlorophyll-proteins were resolved from the thylakoid membranes, or digitonin particles, of a thermophilic cyanobacterium Synechococcus sp. by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Six chlorophyll-proteins with slower electrophoretic mobilities were shown to be P700-chlorophyll a-protein complexes (CP1), whereas faster-moving proteins (CP2) were related to photosystem 2. Extraction of CP1 complexes from the membranes with different detergent/chlorophyll ratios and reelectrophoresis of extracted CP1 complexes indicated that the chlorophyll-proteins are closely interrelated with each other; any CP1 complex could be transformed to other CP1 complexes with faster electrophoretic mobilities. This, together with the Ferguson plot and the polypeptide composition, showed that six CP1 complexes are different in terms of polypeptide composition, oligomerization, SDS-binding, or conformation of the proteins but represent, in the order of increasing electrophoretic mobility, increasing degree of modification of the native P700-chlorophyll a-protein.     

Characterization of a zwf mutant of Synechococcus sp. strain PCC 7942.

A mutant of the cyanobacterium Synechococcus sp. strain PCC 7942 carrying a disrupted gene encoding glucose-6-phosphate dehydrogenase (zwf) produced no detectable glucose-6-phosphate dehydrogenase as assessed by enzyme assay and Western blot (immunoblot) analysis. This mutant exhibited significantly impaired dark viability.     

Acclimation of Arabidopsis thaliana to the light environment: changes in photosynthetic function

Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta was grown under light regimes of differing spectral qualities, which results in differences in the stoichiometries of the two photosynthetic reaction centres. The acclimative value of these changes was investigated by assessing photosynthetic function in these plants when exposed to two spectrally distinct actinic lights. Plants grown in an environment enriched in far-red light were better able to make efficient use of non-saturating levels of actinic light enriched in long-wavelength red light. Simultaneous measurements of chlorophyll fluorescence and absorption changes at 820 nm indicated that differences between plants grown under alternative light regimes can be ascribed to imbalances in excitation of photosystems I and II (PSI, PSII). Measurements of chlorophyll fluorescence emission and excitation spectra at 77 K provided strong evidence that there was little or no difference in the composition or function of PSI or PSII between the two sets of plants, implying that changes in photosynthetic stoichiometry are primarily responsible for the observed differences in photosynthetic function.Abbreviations Chl chlorophyll - FR far-red light - HF highirradiance FR-enriched light (400 mol·m^–2·s^–1, RFR = 0.72) - HW high-irradiance white light (400 mol·m^–2 1·1 s^–1RFR = 1.40) - LHCI, LHCII light-harvesting complex of PSI, PSII - qO quenching of dark-level chlorophyll fluorescence - qN non-photochemical quenching of variable chlorophyll fluorescence - qP photochemical quenching of variable chlorophyll fluorescence - R red light - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase We thank Dr. Sasha Ruban for assistance with the 77 K fluorescence measurements and for helpful discussions. This work was supported by Natural Environment Research Council Grant GR3/7571A.     

The grand design of photosynthesis: Acclimation of the photosynthetic apparatus to environmental cues

Dynamic acclimation of the photosynthetic apparatus in response to environmental cues, particularly light quantity and quality, is a widely-observed and important phenomenon which contributes to the tolerance of plants against stress and helps to maintain, as far as possible, optimal photosynthetic efficiency and resource utilization. This mini-review represents a scrutiny of a number of possible photoreceptors (including the two photosystems acting as light sensors) and signal transducers that may be involved in producing acclimation responses. We suggest that regulation by signal transduction may be effected at each of several possible points, and that there are multiple regulatory mechanisms for photosynthetic acclimation.Abbreviations FR far-red light - LHC I, LHC II light-harvesting chlorophyll a/b-protein complex of PS I and PS II, respectively - P700 primary electron donor of PS I - Pmax maximum photosynthetic capacity - Q_A primary quinone electron acceptor of PS II - q_N, q_P non-photochemical and photochemical quenching, respectively - R red light     

热带雨林三种树苗叶片光合机构对光强的适应

对生长在不同光强(自然日光的8％,25％,50％)下西双版纳热带雨林3种木本植物团花(Anthocephalus chinensis)、玉蕊(Barringtonia pendala)和藤黄(Garrcinia hanburyi)幼苗光合机构的研究表明,随着生长光强的升高,植物叶片的光饱和点、补偿点、净光合速率和非光化学淬灭系数(NPQ)升高,而表现量子效率(AQY)、有效光化学量子产量(Fv／Fm)、光化学淬灭系数(qP)下降．在抗氧化系统中,超氧化物歧化酶(SOD)、抗坏血酸过氧化酶(APX)活性随着光强的升高而升高,而过氢化物酶(CAT)活性与生长光强的变化不一致．抗坏血酸(AsA)含量随着光强的升高而急剧上升。最能反映PFD的变化．可以认为,除与叶黄素循环有关的热耗散增大之外,植物叶片抗氧化系统的加强也是响应强光的一种保护措施．     

The action in vivo of glycine betaine in enhancement of tolerance of Synechococcus sp. strain PCC 7942 to low temperature.

The cyanobacterium Synechococcus sp. strain PCC 7942 was transformed with the codA gene for choline oxidase from Arthrobacter globiformis under the control of a constitutive promoter. This transformation allowed the cyanobacterial cells to accumulate glycine betaine at 60 to 80 mM in the cytoplasm. The transformed cells could grow at 20 degrees C, the temperature at which the growth of control cells was markedly suppressed. Photosynthesis of the transformed cells at 20 degrees C was more tolerant to light than that of the control cells. This was caused by the enhanced ability of the photosynthetic machinery in the transformed cells to recover from low-temperature photoinhibition. In darkness, photosynthesis of the transformed cells was more tolerant to low temperature such as 0 to 10 degrees C than that of the control cells. In parallel with the improvement in the ability of the transformed cells to tolerate low temperature, the lipid phase transition of plasma membranes from the liquid-crystalline state to the gel state shifted toward lower temperatures, although the level of unsaturation of the membrane lipids was unaffected by the transformation. These findings suggest that glycine betaine enhances the tolerance of photosynthesis to low temperature.     

Molecular characterization of a thermostable cyanophycin synthetase from the thermophilic cyanobacterium Synechococcus sp. strain MA19 and in vitro synthesis of cyanophycin and related polyamides.

The thermophilic cyanobacterium Synechococcus sp. strain MA19 contained the structural genes for cyanophycin synthetase (cphA) and cyanophycinase (cphB), which were identified, cloned, and sequenced in this study. The translation products of cphA and cphB exhibited high levels of similarity to corresponding proteins of other cyanobacteria, such as Anabaena variabilis and Synechocystis sp. Recombinant cells of Escherichia coli harboring cphA colinear with lacPO accumulated cyanophycin that accounted for up to 25% (wt/wt) of the dry cell matter in the presence of isopropyl-beta-D-thiogalactopyranoside (IPTG). The cyanophycin synthetase was enriched 123-fold to electrophoretic homogeneity from the soluble fraction of the recombinant cells by anion-exchange chromatography, affinity chromatography, and gel filtration chromatography. The purified cyanophycin synthetase maintained the parental thermophilic character and was active even after prolonged incubation at 50 degrees C; in the presence of ectoine the enzyme retained 90% of its activity even after 2 h of incubation. The in vitro activity of the enzyme depended on ATP, primers, and both substrates, L-arginine and L-aspartic acid. In addition to native cyanophycin, the purified enzyme accepted a modified cyanophycin containing less arginine, alpha-arginyl aspartic acid dipeptide, and poly-alpha,beta-DL-aspartic acid as primers and also incorporated beta-hydroxyaspartic acid instead of L-aspartic acid or L-canavanine instead of L-arginine at a significant rate. The lack of specificity of this thermostable enzyme with respect to primers and substrates, the thermal stability of the enzyme, and the finding that the enzyme is suitable for in vitro production of cyanophycin make it an interesting candidate for biotechnological processes.     

Membrane lipid composition and restoration of photosynthesis during low temperature acclimation in Synechococcus sp. strain PCC 7942

We compared temperature acclimation of the cyanobacterium Synechococcus sp. strain PCC 7942 and two psbA inactivation mutants, R2K1 and R2S2C3, following shifts from 37 to 25°C. Wild-type cultures incubated in the dark at 25°C showed no chill-induction of lipid desaturation, probably because the lipid acclimation is dependent on photosynthesis. Incubation in the light at 25°C, however, induced considerable increases in membrane lipid desaturation, and within 24 h the monoenoic fatty acids increased from about 46 to about 57%. In parallel with this desaturation the ratio of monogalactosyldiacylglycerol to digalactosyldiacylglycerol (MGDG/DGDG) increased. Both of these lipid changes increase the repulsive forces of the hydrophobic chains of the membrane lipids and thereby alter the physical properties of the membrane. As expected, under irradiation this temperature shift also induced a reversible replacement of the constitutive photosystem II protein, D1:1, with an alternative stress form, D1:2. Photosynthesis decreased to 42% of the control level within the initial 2 h of cold incubation, but later recovered. The D1:2 protein accumulated to high levels between 2 and 4 h after the temperature shift, when desaturation of membrane lipids and MGDG/DGDG ratio had not yet increased significantly. Much of this accumulated D1:2 protein was in a higher molecular mass form, termed D1:2^*, which is probably an unprocessed precursor form of the protein. In contrast to the wild-type cells the psbA inactivation mutants, R2K1 and R2S2C3 did not accumulate any precursor form of D1 protein either at the optimal or low growth temperature. The R2S2C3 mutant strain expresses only the constitutive D1:1 protein and suffered severe photoinhibition following the temperature shift. Nevertheless, R2S2C3 eventually recovered some photosynthetic activity, induced lipid desaturation and slowly resumed growth at 25°C, thus demonstrating acclimation to the lower growth temperature. The R2K1 mutant synthesizes only the D1:2 stress form of D1 protein and maintained oxygen evolution at a high level (ca 70% of a control rate) after the low temperature shift. Chill-induced lipid desaturation and increase in MGDG/DGDG ratio did proceed but, for unknown reasons the strain did not resume growth at the lower temperature. The physical properties of the membrane lipids were not the limiting factor for growth resumption. Our results demonstrate that in the wild-type the chill-induced desaturation of membrane lipids follows after the exchange of the two forms of the D1 proteins, but the D1 exchange results in accumulation of unprocessed D1:2^* polypeptides until the lipid composition later acclimates. We also show that the lipid desaturation process in Synechococcus sp. strain PCC 7942 is dependent upon photosynthetic activity.     

Ultrastructure of cell wall and photosynthetic apparatus of the phycobilisome-less Synechocystis sp. strain BO 8402 and phycobilisome-containing derivative strain BO 9201

The ultrastructures of two closely related strains of a novel diazotrophic cyanobacterium, Synechocystis sp. BO 8402 and BO 9201, were examined using ultrathin sections and freeze-fracture electron microscopy. Cells of both strains were surrounded by an unusual thick peptidoglycan layer. Substructures in the layer indicated the presence of microplasmodesmata aligned perpendicular to the free cell surface and in the septum of dividing cells. Synechocystis sp. strain BO 8402 contained lobed, electronopaque, highly fluorescent inclusion bodies consisting of phycocyanin-linker complexes. The thylakoids lacked phycobilisomes and accommodated, in addition to randomly distributed exoplasmic freeze-fracture particles, patches of two-dimensionally ordered arrays of dimeric photosystem II particles in the exoplasmic fracture face. Determination of photosystem I and photosystem II suggested an increase of photosystem II in strain BO 8402. Strain BO 9201 performed phycobilisome-supported photosynthesis and showed rows of dimeric photosystem II particles in the exoplasmic fracture face. Corresponding particle-free grooves in the protoplasmic fracture face were lined by a class of large particles tentatively assigned as trimers of photosystem I. The different lateral organization of protein complexes in the thylakoid membranes and the fine structure of the cell wall are discussed with respect to absorption cross-section of photosynthesis and nitrogen fixation.Abbreviations EF Exoplasmic freeze-fracture face - P 700 Reaction centre chlorophyll of photosystem I - PF Protoplasmic freeze-fracture face - PS I Photosystem I - PS II Photosystem II     

Acclimation of the photosynthetic apparatus and alterations in sugar metabolism in response to inoculation with endophytic fungi

The role of an endophytic Zygomycete Mucor sp. in growth promotion and adaptation of the photosynthetic apparatus to increased energy demands of its hosts Arabidopsis arenosa and Arabidopsis thaliana was evaluated. Inoculation with the fungus improved the water use efficiency of the plants and allowed for them to utilize incident light for photochemistry more effectively by upregulating the expression of several photosystem I‐ and II‐related genes and their respective proteins, proteins involved in light harvesting in PSII and PSI and carbon assimilation. This effect was independent of the ability of the plants to acquire nutrients from the soil. We hypothesize that the accelerated growth of the symbiotic plants resulted from an increase in their demand for carbohydrates and carbohydrate turnover (sink strength) that triggered a simultaneous upregulation of carbon assimilation. Arabidopsis plants inoculated with Mucor sp. exhibited upregulated expression in several genes encoding proteins involved in carbohydrate catabolism, sugar transport, and smaller starch grains that indicate a significant upregulation of carbohydrate metabolism.