RNA helicase, CrhR is indispensable for the energy redistribution and the regulation of photosystem stoichiometry at low temperature in Synechocystis sp. PCC6803

We investigated the role of a cold-inducible and redox-regulated RNA helicase, CrhR, in the energy redistribution and adjustment of stoichiometry between photosystem I (PSI) and photosystem II (PSII), at low temperature in Synechocystis sp. PCC 6803. The results suggest that during low temperature incubation, i.e., when cells are shifted from 34°C to 24°C, wild type cells exhibited light-induced state transitions, whereas the mutant deficient in CrhR failed to perform the same. At low temperature, wild type cells maintained the plastoquinone (PQ) pool in the reduced state due to enhanced respiratory electron flow to the PQ pool, whereas in ?crhR mutant cells the PQ pool was in the oxidized state. Wild type cells were in state 2 and ?crhR cells were locked in state 1 at low temperature. In both wild type and ?crhR cells, a fraction of PSI trimers were changed to PSI monomers. However, in ?crhR cells, the PSI trimer content was significantly decreased. Expression of photosystem I genes, especially the psaA and psaB, was strongly down-regulated due to oxidation of downstream components of PQ in ?crhR cells at low temperature. We demonstrated that changes in the low temperature-induced energy redistribution and regulation of photosystem stoichiometry are acclimatization responses exerted by Synechocystis cells, essentially regulated by the RNA helicase, CrhR, at low temperature.     

RNA解旋酶基因crhR对于集胞藻PCC6803低温适应的作用

蓝藻对低温胁迫的适应涉及许多基因的表达调控,RNA解旋酶基因crhR即是其中之一。研究检查了该基因在集胞藻PCC6803从30℃转到15℃后的转录情况,观察到在2h内有瞬时诱导表达。该基因失活导致集胞藻在15℃下几乎不能生长,光合作用和呼吸速率大幅下降,脂质过氧化物不能被有效清除。以PrbcL过量表达crhR基因可互补突变株表型,并且在低温下生长略优于野生型。在野生型中,低温诱导脂肪酸脱饱和酶基因desA、desB、desD表达上调,膜脂不饱和度增加;而在crhR突变株中,低温诱导的desB基因的上调表达显著削弱,同时多不饱和脂肪酸含量没有显著增加。推测crhR基因可能影响蓝藻在低温胁迫下的蛋白合成或通过伴侣蛋白发挥影响。       

High CO2 concentration alleviates the block in photosynthetic electron transport in an ndhB-inactivated mutant of Synechococcus sp. PCC 7942.

The high-concentration CO2-requiring mutant N5 of Synechococcus sp. PCC 7942 was obtained by the insertion of a kanamycin-resistant gene at the EcoRI site, 12.4 kb upstream of rbc. The mutant is unable to accumulate inorganic carbon internally and exhibits very low apparent photosynthetic affinity for inorganic carbon but a photosynthetic Vmax similar to that of the wild type. Sequence and northern analyses showed that the insertion inactivated a gene highly homologous to ndhB, encoding subunit II of NADH dehydrogenase in Synechocystis sp. PCC 6803 (T. Ogawa [1991] Proc Natl Acad Sci USA 88: 4275-4279). When the mutant and the wild-type cells were exposed to 5% CO2 in air, their photosynthetic electron transfer capabilities, as revealed by fluorescence and thermoluminescence measurements, were similar. On the other hand, a significant decrease in variable fluorescence was observed when the mutant (but not the wild-type) cells were exposed to low CO2 under continuous light. The same treatment also resulted in a shift (from 38-27 degrees C) in the temperature at which the maximal thermoluminescence emission signal was obtained in the mutant but not in the wild type. These results may indicate that subunit II of NADH dehydrogenase is essential for the functional operation of the photosynthetic electron transport in Synechococcus under low but not high levels of CO2. We suggest that the inability to accumulate inorganic carbon under air conditions stems from disrupture of electron transport in this mutant.     

Characterization of a mutant lacking carboxysomal carbonic anhydrase from the cyanobacterium Synechocystis PCC6803

A fully-segregated mutant (ccaA::kanR) defective in the ccaA gene, encoding a carboxysome-associated beta-carbonic anhydrase (CA), was generated in the cyanobacterium Synechocystis sp. PCC6803 by insertional mutagenesis. Immunoblot analysis indicated that the CcaA polypeptide was absent from the carboxysome-enriched fraction obtained from ccaA::kanR, but was present in wild-type (WT) cells. The carboxysome-enriched fraction isolated from WT cells catalyzed 18O exchange between 13C18O2 and H2O, indicative of CA activity, while ccaA::kanR carboxysomes did not. Transmission and immunogold electron microscopy revealed that carboxysomes of WT and ccaA::kanR were of similar size, shape and cellular distribution, and contained most of the cellular complement of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The ccaA::kanR cells were substantially smaller than WT and were unable to grow autotrophically at air levels of CO2. However, cell division occurred at near-WT rates when ccaA::kanR was supplied with 5% CO2 (v/v) in air. The apparent photosynthetic affinity of the mutant for inorganic carbon (Ci) was 500-fold lower than that of WT cells although intracellular Ci accumulation was comparable to WT measurements. Mass spectrometric analysis revealed that the CA-like activity associated with the active CO2 transport system was retained by ccaA::kanR cells and was inhibited by H2S, indicating that CO2 transport was distinct from the CcaA-mediated dehydration of intracellular HCO3-. The data suggest that the ccaA mutant was unable to efficiently utilize the internal Ci pool for carbon fixation and that the high-CO2-requiring phenotype of ccaA::kanR was due primarily to an inability to generate enough CO2 in the carboxysomes to sustain normal rates of photosynthesis.     

干出和沉水不同条件下羊栖菜光合作用碳源获得机制比较(英文)

经济海洋褐藻羊栖菜(Hizikia fusiforme(Harv.)Okamura)低潮时常常周期性地暴露于空气中。为了认识这种海藻在潮汐循环背景下的光合特征,对其在高潮沉水和低潮干出不同条件下的光合作用碳素获得机制进行了比较。沉水时,羊栖菜主要利用海水中HCO_3~-作为外源无机碳源驱动光合作用;而在干出条件下,其光合作用的主要碳源为空气中的CO_2。在这两种不同环境条件下,光合作用与pH值的关系不同:沉水状态时,羊栖菜在高pH值(10.0)下光合活性很弱;而在干出条件下,羊栖菜在高pH值时仍有较高的光合活性。然而,光合作用无论是在沉水还是在干出条件下,对外源碳源的获得都表现出对胞外碳酸酐酶(CA)强烈的依赖性,并且其光合速率都受周围环境中无机碳源水平的限制。此外,在沉水和干出两种环境条件下,羊栖菜光合作用都表现出对氧气的敏感性。这表明,在羊栖菜中,依赖胞外CA的碳源获得机制不能使细胞内CO_2浓度提高到阻碍其光呼吸的程度。增加空气中或海水中无机碳的浓度,能促进羊栖菜的光合作用,进而增加这种海藻的水产养殖产量。     

干出和沉水不同条件下羊栖菜光合作用碳源获得机制比较

经济海洋褐藻羊栖菜(Hizikia fusiforme(Harv.)Okamura)低潮时常常周期性地暴露于空气中.为了认识这种海藻在潮汐循环背景下的光合特征,对其在高潮沉水和低潮干出不同条件下的光合作用碳素获得机制进行了比较.沉水时,羊栖菜主要利用海水中HCO3-作为外源无机碳源驱动光合作用;而在干出条件下,其光合作用的主要碳源为空气中的CO2.在这两种不同环境条件下,光合作用与pH值的关系不同:沉水状态时,羊栖菜在高pH值(10.0)下光合活性很弱;而在干出条件下,羊栖菜在高pH值时仍有较高的光合活性.然而,光合作用无论是在沉水还是在干出条件下,对外源碳源的获得都表现出对胞外碳酸酐酶(CA)强烈的依赖性,并且其光合速率都受周围环境中无机碳源水平的限制.此外,在沉水和干出两种环境条件下,羊栖菜光合作用都表现出对氧气的敏感性.这表明,在羊栖菜中,依赖胞外CA的碳源获得机制不能使细胞内CO2浓度提高到阻碍其光呼吸的程度.增加空气中或海水中无机碳的浓度,能促进羊栖菜的光合作用,进而增加这种海藻的水产养殖产量.     

The response of the high altitude C(4) grass Muhlenbergia montana (Nutt.) A.S. Hitchc. to long- and short-term chilling.

The acclimation of C(4) photosynthesis to low temperature was studied in the montane grass Muhlenbergia montana in order to evaluate inherent limitations in the C(4) photosynthetic pathway following chilling. Plants were grown in growth cabinets at 26 degrees C days, but at night temperatures of either 16 degrees C (the control treatment), 4 degrees C for at least 28 nights (the cold-acclimated treatment), or 1 night (the cold-stress treatment). Below a measurement temperature of 25 degrees C, little difference in the thermal response of the net CO(2) assimilation rate (A) was observed between the control and cold-acclimated treatment. By contrast, above 30 degrees C, A in the cold-acclimated treatment was 10% greater than in the control treatment. The temperature responses of Rubisco activity and net CO(2) assimilation rate were similar below 22 degrees C, indicating high metabolic control of Rubisco over the rate of photosynthesis at cool temperatures. Analysis of the response of A to intercellular CO(2) level further supported a major limiting role for Rubisco below 20 degrees C. As temperature declined, the CO(2) saturated plateau of A exhibited large reductions, while the initial slope of the CO(2) response was little affected. This type of response is consistent with a Rubisco limitation, rather than limitations in PEP carboxylase capacity. Stomatal limitations at low temperature were not apparent because photosynthesis was CO(2) saturated below 23 degrees C at air levels of CO(2). In contrast to the response of photosynthesis to temperature and CO(2) in plants acclimated for 4 weeks to low night temperature, plants exposed to 4 degrees C for one night showed substantial reduction in photosynthetic capacity at temperatures above 20 degrees C. Because these reductions were at both high and low CO(2), enzymes associated with the C(4) carbon cycle were implicated as the major mechanisms for the chilling inhibition. These results demonstrate that C(4) plants from climates with low temperature during the growing season can fully acclimate to cold stress given sufficient time. This acclimation appears to involve reversal of injury to the C(4) cycle following initial exposure to low temperature. By contrast, carbon gain at low temperatures generally appears to be constrained by the carboxylation capacity of Rubisco, regardless of acclimation time. The inability to overcome the Rubisco limitation at low temperature may be an inherent limitation restricting C(4) photosynthetic performance in cooler climates.     

Source-sink imbalance increases with growth temperature in the spring geophyte Erythronium americanum

Spring geophytes produce larger storage organs and present delayed leaf senescence under lower growth temperature. Bulb and leaf carbon metabolism were investigated in Erythronium americanum to identify some of the mechanisms that permit this improved growth at low temperature. Plants were grown under three day/night temperature regimes: 18/14 °C, 12/8 °C, and 8/6 °C. Starch accumulated more slowly in the bulb at lower temperatures probably due to the combination of lower net photosynthetic rate and activation of a 'futile cycle' of sucrose synthesis and degradation. Furthermore, bulb cell maturation was delayed at lower temperatures, potentially due to the delayed activation of sucrose synthase leading to a greater sink capacity. Faster starch accumulation and the smaller sink capacity that developed at higher temperatures led to early starch saturation of the bulb. Thereafter, soluble sugars started to accumulate in both leaf and bulb, most probably inducing decreases in fructose-1,6-bisphosphatase activity, triose-phosphate utilization in the leaf, and the induction of leaf senescence. Longer leaf life span and larger bulbs at lower temperature appear to be due to an improved equilibrium between carbon fixation capacity and sink strength, thereby allowing the plant to sustain growth for a longer period of time before feedback inhibition induces leaf senescence.     

Trienoic fatty acids are required to maintain chloroplast function at low temperatures

The chloroplast membranes of all higher plants contain very high proportions of trienoic fatty acids. To investigate how these lipid structures are important in photosynthesis, we have generated a triple mutant line of Arabidopsis that contains negligible levels of trienoic fatty acids. For mutant plants grown at 22 degrees C, photosynthetic fluorescence parameters were indistinguishable from wild type at 25 degrees C. Lowering the measurement temperature led to a small decrease in photosynthetic quantum yield, Phi(II), in the mutant relative to wild-type controls. These and other results indicate that low temperature has only a small effect on photosynthesis in the short term. However, long-term growth of plants at 4 degrees C resulted in decreases in fluorescence parameters, chlorophyll content, and thylakoid membrane content in triple-mutant plants relative to wild type. Comparisons among different mutant lines indicated that these detrimental effects of growth at 4 degrees C are strongly correlated with trienoic fatty acid content with levels of 16:3 + 18:3, approximately one-third of wild type being sufficient to sustain normal photosynthetic function. In total, our results indicate that trienoic fatty acids are important to ensure the correct biogenesis and maintenance of chloroplasts during growth of plants at low temperatures.     

The carbon gain benefits of solar tracking in a desert annual

Abstract. A comparison between two sympatric winter desert annuals, Camissonia claviformis and Malvastrum rotundi folium showed that both gained similar amounts of carbon during a spring day, although by very different means. Camissonia has horizontally fixed leaves which have a very high photosynthetic capacity. The temperature optimum of photosynthesis for this species is near 20°C. Malvastrum has leaves with a lower photosynthetic capacity and a photosynthetic temperature optimum near 30°C. Leaves of the latter species remain normal to the sun throughout the course of the day. The tracking response and high temperature optimum for photosynthesis of Malvastrum result in a high daily carbon gain and also a high water-use efficiency.     

The effects of severe carbon limitation on the green seaweed,Ulva conglobata (Chlorophyta)

Low inorganic carbon (Ci) concentrations in seawater are usually an important factor controlling photosynthesis and growth of seaweeds. The green seaweed, Ulva conglobata Kjellm, collected from a rock pool in a middle intertidal zone located at Nanao Island, Shantou, China, were cultured under low Ci level for several days, to examine the effect of severe carbon limitation on photosynthesis. The rather high pH compensation points obtained from the pH-drift experiments indicated that U. conglobata was capable of acquiring HCO₃ ^? from surrounding seawater as its Ci source for photosynthesis. However, thalli of U. conglobata cultured in Ci-starved seawater exhibited a decline of biomass, showing that the realistic photosynthetic carbon gain could not compensate for the respiratory carbon consumption in the thalli under severe Ci limitation during laboratory culture. Compared with ambient Ci conditions, the culture under severe Ci limitation significantly had an increased pigment content, but a lower maximum quantum yield and photosynthetic electron transport rate. Additionally, the maximum carbon-saturating photosynthesis rate and the apparent photosynthetic conductance of U. conglobata thalli increased in cultures with severe Ci limitation compared with ambient Ci in low N-grown thalli. The results suggest that under severe Ci limitation, U. conglobata thalli increased capacities of both light absorption processes and carbon fixation pathways.     

Meeting the photosynthetic demand for inorganic carbon in an alga-invertebrate association: preferential use of CO2 by symbionts in the giant clam Tridacna gigas

Unlike most marine invertebrates which excrete respiratory CO2, giant clams (Tridacna gigas) must acquire inorganic carbon (Ci) in order to support their symbiotic population of photosynthetic dinoflagellates. Their capacity to meet this demand will be reflected in the Ci concentration of their haemolymph during periods of high photosynthesis. The Ci concentration in haemolymph was found to be inversely proportional to irradiance with a minimum Ci concentration of 0.75 mM at peak light levels increasing to 1.2 mM in the dark. The photosynthetic rate of isolated zooxanthellae under conditions that prevail in the haemolymph at peak light levels was significantly less than the potential Pmax (maximum photosynthetic rate) indicating that zooxanthellae are carbon limited in hospite. This is consistent with previous studies on the hermatypic coral symbiosis. The Pmax was not affected by pH but there was a dramatic increase in the half-saturation constant for Ci (K0.5 (Ci)) with increasing pH (6.5-9.0) and only a small decrease in K0.5 (CO2) over the same range. These results indicate that zooxanthellae in giant clams use CO2 as the primary source of their Ci in contrast to symbionts in corals, which use bicarbonate. The physiological implications are discussed and comparison is made with the coral symbiosis.     

Modelling photosynthetic responses to temperature of grapevine (Vitis vinifera cv. Semillon) leaves on vines grown in a hot climate

Field measurements of photosynthesis of Vitis vinifera cv. Semillon leaves in relation to a hot climate, and responses to photon flux densities (PFDs) and internal CO(2) concentrations (c(i) ) at leaf temperatures from 20 to 40 °C were undertaken. Average rates of photosynthesis measured in situ decreased with increasing temperature and were 60% inhibited at 45 °C compared with 25 °C. This reduction in photosynthesis was attributed to 15-30% stomatal closure. Light response curves at different temperatures revealed light-saturated photosynthesis optimal at 30 °C but also PFDs saturating photosynthesis increased from 550 to 1200 μmol (photons) m(-2)s(-1) as temperatures increased. Photosynthesis under saturating CO(2) concentrations was optimal at 36 °C while maximum rates of ribulose 1,5-bisphosphate (RuBP) carboxylation (V(cmax)) and potential maximum electron transport rates (J(max)) were also optimal at 39 and 36 °C, respectively. Furthermore, the high temperature-induced reduction in photosynthesis at ambient CO(2) was largely eliminated. The chloroplast CO(2) concentration at the transition from RuBP regeneration to RuBP carboxylation-limited assimilation increased steeply with an increase in leaf temperature. Semillon assimilation in situ was limited by RuBP regeneration below 30 °C and above limited by RuBP carboxylation, suggesting high temperatures are detrimental to carbon fixation in this species.     

The Chlamydomonas reinhardtii cia3 mutant lacking a thylakoid lumen-localized carbonic anhydrase is limited by CO2 supply to rubisco and not photosystem II function in vivo

The Chlamydomonas reinhardtii cia3 mutant has a phenotype indicating that it requires high-CO(2) levels for effective photosynthesis and growth. It was initially proposed that this mutant was defective in a carbonic anhydrase (CA) that was a key component of the photosynthetic CO(2)-concentrating mechanism (CCM). However, more recent identification of the genetic lesion as a defect in a lumenal CA associated with photosystem II (PSII) has raised questions about the role of this CA in either the CCM or PSII function. To resolve the role of this lumenal CA, we re-examined the physiology of the cia3 mutant. We confirmed and extended previous gas exchange analyses by using membrane-inlet mass spectrometry to monitor(16)O(2),(18)O(2), and CO(2) fluxes in vivo. The results demonstrate that PSII electron transport is not limited in the cia3 mutant at low inorganic carbon (Ci). We also measured metabolite pools sizes and showed that the RuBP pool does not fall to abnormally low levels at low Ci as might be expected by a photosynthetic electron transport or ATP generation limitation. Overall, the results demonstrate that under low Ci conditions, the mutant lacks the ability to supply Rubisco with adequate CO(2) for effective CO(2) fixation and is not limited directly by any aspect of PSII function. We conclude that the thylakoid CA is primarily required for the proper functioning of the CCM at low Ci by providing an ample supply of CO(2) for Rubisco.     

Conditional,Temperature-Induced Proteolytic Regulation of Cyanobacterial RNA Helicase Expression

Conditional proteolysis is a crucial process regulating the abundance of key regulatory proteins associated with the cell cycle, differentiation pathways, or cellular response to abiotic stress in eukaryotic and prokaryotic organisms. We provide evidence that conditional proteolysis is involved in the rapid and dramatic reduction in abundance of the cyanobacterial RNA helicase, CrhR, in response to a temperature upshift from 20 to 30°C. The proteolytic activity is not a general protein degradation response, since proteolysis is only present and/or functional in cells grown at 30°C and is only transiently active at 30°C. Degradation is also autoregulatory, since the CrhR proteolytic target is required for activation of the degradation machinery. This suggests that an autoregulatory feedback loop exists in which the target of the proteolytic machinery, CrhR, is required for activation of the system. Inhibition of translation revealed that only elongation is required for induction of the temperature-regulated proteolysis, suggesting that translation of an activating factor was already initiated at 20°C. The results indicate that Synechocystis responds to a temperature shift via two independent pathways: a CrhR-independent sensing and signal transduction pathway that regulates induction of crhR expression at low temperature and a CrhR-dependent conditional proteolytic pathway at elevated temperature. The data link the potential for CrhR RNA helicase alteration of RNA secondary structure with the autoregulatory induction of conditional proteolysis in the response of Synechocystis to temperature upshift.     

Carbon concentration mechanisms in photosynthetic microorganisms

Unicellular green algae and cyanobacteria have mechanism to actively concentrate dissolved inorganic carbon into the cells, only if they are grown with air levels of CO2. The carbon concentration mechanisms are commonly known as "CCM" or "DIC-pumps". The DIC-pumps are environmental adaptation that function to actively transport and accumulate inorganic carbon (HCO3- and CO2; Ci) within the cell and then uses this Ci pool to actively increase the concentration of CO2 at the site of ribulose bisphosphate carboxylase-oxygenase (Rubisco), the primary CO2-fixing enzyme. The current working model for dissolved inorganic carbon concentration mechanism in unicellular green algae includes several isoforms of carbonic anhydrase (CA), and ATPase driven active transporters at the plasmalemma and at the inner chloroplast envelopes. In the past fifteen years, significant progress has been made in isolating and characterizing the various isoforms of carbonic anhydrase at the biochemical and molecular level. However, we have an inadequate understanding of active transporters that are located on the plasmalemma and at the chloroplast envelopes. In this mini-review we focus on certain aspects of the induction, function and significance of the dissolved inorganic carbon concentration mechanisms in aquatic photosynthetic microorganisms.     

沉水植物光合作用的特点与研究进展

沉水植物属于高等植物,由陆生被子植物演化而来,它们在形态、光合生态生理方面对水下生活环境发生了一系列适应性变化。沉水植物的光合作用受水体中光、温度、pH和无机碳等影响,本文对此进行了综述。水中低CO2扩散率以及细胞外较厚的扩散层阻碍了沉水植物净碳的吸收,因此,沉水植物光合作用速率受到无机碳供应的限制。为获得无机碳,沉水植物在形态结构和生理生化上表现一定的特性,包括薄的叶片层并含有叶绿体以及对HCO3-利用的能力,拟C4型和CAM型光合代谢途径的选择。这些是沉水植物碳浓缩机制的具体体现。     

Characterization of a Photosynthetic Mutant Strain of Chlamydomonas reinhardi Deficient in Phosphoribulokinase Activity

A mutant strain of the unicellular green alga, Chlamydomonas reinhardi, is unable to fix carbon dioxide by photosynthesis because it is deficient in phosphoribulokinase activity. The absence of light-dependent carbon dioxide fixation in cells of the mutant strain supports the operation of the Calvin-Benson scheme of photosynthetic carbon dioxide fixation in this organism. No deficiency other than low phosphoribulokinase activity was found which would account for the inability of cells of the mutant strain to fix carbon dioxide by photosynthesis. Activities comparable to those in the wild-type strain were found for eight other enzymes of the Calvin cycle and two enzymes associated with the C₄ dicarboxylic acid pathway. The normal rates of nicotinamide adenine dinucleotide phosphate photoreduction and of photosynthetic phosphorylation observed in chloroplast fragments prepared from cells of the mutant strain indicated that the photosynthetic electron transport chain in the mutant is intact.