Convergence and divergence of the photoregulation of pigmentation and cellular morphology in Fremyella diplosiphon

Photosynthetic pigment accumulation and cellular and filament morphology are regulated reversibly by green light (GL) and red light (RL) in the cyanobacterium Fremyella diplosiphon during complementary chromatic adaptation (CCA). The photoreceptor RcaE (regulator of chromatic adaptation), which appears to function as a light-responsive sensor kinase, controls both of these responses. Recent findings indicate that downstream of RcaE, the signaling pathways leading to light-dependent changes in morphology or pigment synthesis and/or accumulation branch, and utilize distinct molecular components. We recently reported that the regulation of the accumulation of the GL-absorbing photosynthetic accessory protein phycoerythrin (PE) and photoregulation of cellular morphology are largely independent, as many mutants with severe PE accumulation defects do not have major disruptions in the regulation of cellular morphology. Furthermore, morphology can be disrupted under GL without impacting GL-dependent PE accumulation. Most recently, however, we determined that the disruption of the cpeR gene, which encodes a protein that is known to function as an activator of PE synthesis under GL, results in disruption of cellular morphology under GL and RL. Thus, apart from RcaE, CpeR is only the second known regulator to impact morphology under both light conditions in F. diplosiphon.     

Characterization of green mutants in Fremyella diplosiphon provides insight into the impact of phycoerythrin deficiency and linker function on complementary chromatic adaptation

Functions of phycobiliprotein (PBP) linkers are less well studied than other PBP polypeptides that are structural components or required for the synthesis of the light-harvesting phycobilisome (PBS) complexes. Linkers serve both structural and functional roles in PBSs. Here, we report the isolation of a phycoerythrin (PE) rod-linker mutant and a novel PE-deficient mutant in Fremyella diplosiphon. We describe their phenotypic characterization, including light-dependent photosynthetic pigment accumulation and photoregulation of cellular morphology. PE-linker protein CpeE and a novel protein impact PE accumulation, and thus PBS function, primarily under green light conditions.     

A novel role for a TonB-family protein and photoregulation of iron acclimation in Fremyella diplosiphon

Photosynthetic organisms display adaptations to changes in light and nutrient availability. Iron, which is required for the function of photosynthetic photosystems and other important biochemical processes, is an essential mineral that consequently impacts not only overall photosynthetic efficiency, but also the physiology of organisms in general. Our recent study represents the first functional characterization of a cyanobacterial TonB protein. TonB proteins classically are membrane proteins that support the transport of iron and vitamin B12 into cells. TonB proteins thus generally serve a critical role in organismal iron acclimation. We recently identified FdTonB, a TonB-family protein, in the filamentous freshwater cyanobacterium Fremyella diplosiphon. FdTonB contains conserved TonB residues and domains, as well as novel protein domains. Our recent study, however, supports a novel function for this protein in the photoregulation of morphology, rather than iron acclimation, in F. diplosiphon. Our detailed investigations into the responses of SF33 wild-type and ΔtonB mutant strains did not support a role for FdTonB in organismal responses to iron limitation. However, close examination of our recent results did highlight a novel interaction between light and iron acclimation in F. diplosiphon.Key words: cellular morphology, complementary chromatic adaptation, cyanobacteria, iron, phycobiliprotein, photomorphogenesis, photoregulation, siderophores     

CpcF-dependent regulation of pigmentation and development in Fremyella diplosiphon

Cyanobacteria harvest light for photosynthesis using photosynthetic light-harvesting complexes called phycobilisomes (PBSs). Lyases are enzymes responsible for covalent attachment of light-absorbing chromophores to the phycobiliproteins (PBPs) contained in PBSs. We isolated a pigmentation mutant in the filamentous cyanobacterium Fremyella diplosiphon and determined that it possesses an insertional mutation in cpcF, which encodes one component of a heterodimeric phycocyanin lyase. Here, we discuss the implications of the mutation in cpcF on light-dependent pigmentation and morphology responses characteristic of complementary chromatic adaptation in F. diplosiphon. Although cpcF encodes a phycocyanin lyase, significant decreases in the levels of all classes of PBPs are associated with CpcF deficiency in F. diplosiphon. Notably, CpcF deficiency has a limited effect on the shape of F. diplosiphon cells, but significantly impacts filament length. Possible mechanisms for the broad impact of CpcF deficiency on pigmentation and filament morphology are discussed.     

Temporal dynamics of changes in reactive oxygen species (ROS) levels and cellular morphology are coordinated during complementary chromatic acclimation in Fremyella diplosiphon

Fremyella diplosiphon alters the phycobiliprotein composition of its light-harvesting complexes, i.e., phycobilisomes, and its cellular morphology in response to changes in the prevalent wavelengths of light in the external environment in a phenomenon known as complementary chromatic acclimation (CCA). The organism primarily responds to red light (RL) and green light (GL) during CCA to maximize light absorption for supporting optimal photosynthetic efficiency. Recently, we found that RL-characteristic spherical cell morphology is associated with higher levels of reactive oxygen species (ROS) compared to growth under GL where lower ROS levels and rectangular cell shape are observed. The RL-dependent association of increased ROS levels with cellular morphology was demonstrated by treating cells with a ROS-scavenging antioxidant which resulted in the observation of GL-characteristic rectangular morphology under RL. To gain additional insights into the involvement of ROS in impacting cellular morphology changes during CCA, we conducted experiments to study the temporal dynamics of changes in ROS levels and cellular morphology during transition to growth under RL or GL. Alterations in ROS levels and cell morphology were found to be correlated with each other at early stages of acclimation of low white light-grown cells to growth under high RL or cells transitioned between growth in RL and GL. These results provide further general evidence that significant RL-dependent increases in ROS levels are temporally correlated with changes in morphology toward spherical. Future studies will explore the light-dependent mechanisms by which ROS levels may be regulated and the direct impacts of ROS on the observed morphology changes.     

<Emphasis Type="Italic">Fremyella diplosiphon</Emphasis> as a Biodiesel Agent: Identification of Fatty Acid Methyl Esters via Microwave-Assisted Direct In Situ Transesterification

Increasing concerns on environmental and economic issues linked to fossil fuel use has driven great interest in cyanobacteria as third-generation biofuel agents. In this study, the biodiesel potential of a model photosynthetic cyanobacterium, Fremyella diplosiphon, was identified by fatty acid methyl esters (FAME) via direct transesterification. Total lipids in wild type (Fd33) and halotolerant (HSF33-1 and HSF33-2) strains determined by gravimetric analysis yielded 19% cellular dry weight (CDW) for HSF33-1 and 20% CDW for HSF33-2, which were comparable to Fd33 (18% CDW). Gas chromatography-mass spectrometry detected a high ratio of saturated to unsaturated FAMEs (2.48–2.61) in transesterified lipids, with methyl palmitate being the most abundant (C16:0). While theoretical biodiesel properties revealed high cetane number and oxidative stability, high cloud and pour point values indicated that fuel blending could be a viable approach. Significantly high FAME abundance in total transesterified lipids of HSF33-1 (40.2%) and HSF33-2 (69.9%) relative to Fd33 (25.4%) was identified using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry, indicating that robust salt stress response corresponds to higher levels of extractable FAME. Alkanes, a key component in conventional fuels, were present in F. diplosiphon transesterified lipids across all strains confirming that natural synthesis of these hydrocarbons is not inhibited during biodiesel production. While analysis of photosynthetic pigments and phycobiliproteins did not reveal significant differences, FAME abundance varied significantly in wild type and halotolerant strains indicating that photosynthetic pathways are not the sole factors that determine fatty acid production. We characterize the potential of F. diplosiphon for biofuel production with FAME yields in halotolerant strains higher than the wild type with no loss in photosynthetic pigmentation.     

Piriformospora indica?rescues growth diminution of rice seedlings during high salt stress

Piriformospora indica association has been reported to increase biotic as well as abiotic stress tolerance of its host plants. We analyzed the beneficial effect of P. indica association on rice seedlings during high salt stress conditions (200 and 300 mM NaCl). The growth parameters of rice seedlings such as root and shoot lengths or fresh and dry weights were found to be enhanced in P. indica-inoculated rice seedlings as compared with non-inoculated control seedlings, irrespective of whether they are exposed to salt stress or not. However, salt-stressed seedlings performed much better in the presence of the fungus compared with non-inoculated control seedlings. The photosynthetic pigment content [chlorophyll (Chl) a, Chl b, and carotenoids] was significantly higher in P. indica-inoculated rice seedlings under high salt stress conditions as compared with salt-treated non-inoculated rice seedlings, in which these pigments were found to be decreased. Proline accumulation was also observed during P. indica colonization, which may help the inoculated plants to become salt tolerant. Taken together, P. indica rescues growth diminution of rice seedlings under salt stress.     

集球藻对盐胁迫的生理适应及细胞结构变化

盐胁迫是影响荒漠区土壤藻类生存的重要环境因子。集球藻是一种广泛分布于生物土壤结皮中的球状绿藻, 能够积累红色素(如虾青素)和油滴, 显示出其独特的生理特性和潜在的应用价值。目前对集球藻的生理、细胞结构以及色素积累的研究非常匮乏。以从荒漠生物结皮中分离的一种集球藻为材料, 在实验室条件下研究盐胁迫对集球藻生物量、光合活性、膜脂过氧化产物丙二醛含量、细胞可溶性蛋白质含量和可溶性糖含量以及细胞结构的影响。研究结果表明, 与对照处理相比, 盐胁迫导致集球藻生物量和光合活性的显著降低, 细胞可溶性蛋白和可溶性糖呈现一定的积累。同时盐处理导致集球藻膜脂丙二醛含量大量增加, SOD和CAT 活性升高。研究还表明, 对照处理下细胞结构完整, 细胞器形态清晰, 生长后期有大量脂肪体积累。在盐处理下藻体细胞形态结构出现阶段性破坏特征和脂肪体以及淀粉粒的积累, 此外细胞器结构模糊和消失, 细胞出现质壁分离和空泡化等。研究为更好地揭示集球藻在盐胁迫环境中的生理适应特性、微结构特征以及色素积累机制具有重要的科学意义, 并为该藻的基础和应用研究提供实验资料。       

Physiological responses of Plantago algarbiensis and P. almogravensis shoots and plantlets to low pH and aluminum stress

We investigated the impact of low pH and aluminum (Al) stress on the growth, nutrients concentration, chlorophyll a fluorescence, photosynthetic pigment contents, proline and carbohydrate accumulation in shoots and plantlets (leaves and roots) of Plantago almogravensis and P. algarbiensis. Both species accumulated considerable and similar amounts of Al in their tissues, mainly in the roots. The presence of Al caused a significant reduction on root elongation in P. algarbiensis. Low pH and Al induced significant changes on nutrient accumulation, but no significant alterations on the maximum efficiency of PSII (F _v/F _m), quantum yield of PSII photochemistry (?_PSII), quantum yield of regulated energy dissipation (?_NPQ) and quantum yield of non-regulated energy dissipation (?_NO) were detected in both species in response to these stresses. However, Al increased significantly the non-photochemical quenching and the chlorophyll b content and decreased the PSII excitation pressure (1 ? q _p) in P. almogravensis leaves. Both stress treatments induced carbohydrate accumulation in the shoots and roots of this species, but not in leaves. In P. algarbiensis, low pH and Al decreased the photosynthetic pigment contents in the shoots, whereas Al stimulated the carbohydrate accumulation in the leaves. Although our data showed that both species are tolerant to Al³⁺ and H⁺, P. almogravensis appeared to be more adapted to maintain cellular physiology and growth under those conditions.     

Osmoregulation and cell composition in salt-adaptation of Nostoc muscorum

Adaptation to salt in the cyanobacterium Nostocmuscorum, is composed of a few mechanisms which together lead to the generation of a salt-tolerant cell. The initial mechanism combines a stimulation of photosynthetic activity with the accumulation of sucrose as an osmoregulator. The secondary mechanism involves the adaptation of N₂ fixation activity and protein biosynthesis. The adaptation is most efficient in response to NaCl-induced stress and functions only partially under stress induced by either KCl or a nonionic osmoticum such as mannitol.     

Salt adaptation of the cyanobacterium synechococcus 6311 growing in a continuous culture (turbidostat)

Physiological aspects of salt-adaptation in the cyanobacterium Synechococcus 6311 growing in a continuous culture (turbidostat) were studied. The process of salt-adaptation was completed within 3 days, as expressed by the specific growth rate of cells grown in the presence of 0.2 and 0.4 molar NaCl. An increase in photosynthetic activity during the adaptation period leads to the accumulation of soluble sugars, essential for osmoregulation in the salt-grown cells. Cells grown in the presence of 0.4 molar NaCl showed an initial inhibition in the rate of protein synthesis which was enhanced after the 1st day of salt stress. After adaptation, salt-grown cells showed an increase in phycobiliprotein synthesis and a higher phycobiliprotein to protein ratio.     

Differences in growth,pigment composition and photosynthetic rates of two phenotypes Microcystis aeruginosa strains under high and low iron conditions

This paper provided insight into the influence of iron on the growth of Microcystis aeruginosa strains related to different phenotypes of this species. In this research it was intended to compare the growth, pigment composition, photosynthetic efficiency and extracellular polysaccharides production of unicellular and colonial strains of M. aeruginosa. A significantly growth inhibition under iron-limited condition on unicellular M. aeruginosa was noted, whereas the colonial strain could maintain a steady growth along with the culture time. This observation was reconfirmed by the content of chlorophyll a. Compared with unicellular strain; the colonial strain exhibited a higher PSII maximum light energy transformation, photosynthetic oxygen evolution and extracellular polysaccharides (EPS) production in iron-limited condition. Further, in order to gain more information about the accessibility of iron in the two phenotypic Microcystis, we found the two strains could produce hydroxamate-type siderophores, the content of siderophores produced by the colonial strain was more than those in unicellular strain under the iron-limited condition. It was interpreted as an adaptation to the dilute environment. Our results demonstrated that the colonial phenotypes possessed stronger ability to endure iron-limited condition than unicellular strain by higher pigment contents, higher photosynthetic activities, higher EPS production and higher siderophores secretion. It might elucidate that the colonial M. aeruginosa bloom can sustain in eutrophic reservoirs and lakes.     

ROS accumulation and oxidative damage to cell structures in Saccharomyces cerevisiae wine strains during fermentation of high-sugar-containing medium

To further elucidate the impact of fermentative stress on Saccharomyces cerevisiae wine strains, we have here evaluated markers of oxidative stress, oxidative damage and antioxidant response in four oenological strains of S. cerevisiae, relating these to membrane integrity, ethanol production and cell viability during fermentation in high-sugar-containing medium. The cells were sampled at different fermentation stages and analysed by flow cytometry to evaluate membrane integrity and accumulation of reactive oxygen species (ROS). At the same time, catalase and superoxide dismutase activities, trehalose accumulation, and protein carbonylation and degradation were measured. The results indicate that the stress conditions occurring during hypoxic fermentation in high-sugar-containing medium result in the production of ROS and trigger an antioxidant response. This involves superoxide dismutase and trehalose for the protection of cell structures from oxidative damage, and protein catabolism for the removal of damaged proteins. Cell viability, membrane integrity and ethanol production depend on the extent of oxidative damage to cellular components. This is, in turn, related to the ‘fitness’ of each strain, which depends on the contribution of individual cells to ROS accumulation and scavenging. These findings highlight that the differences in individual cell resistances to ROS contribute to the persistence of wine strains during growth under unfavourable culture conditions, and they provide further insights into our understanding of yeast behaviour during industrial fermentation.     

Low-molecular mass carbohydrate accumulation in cyanobacteria from a marine microbial mat in response to salt

Forty seven strains of cyanobacteria, all isolated from microbial mats of intertidal sediments of the island of Mellum (North Sea), were analyzed for the presence of organic osmotica. The cyanobacteria examined belonged to taxonomically different groups and were classified according to their salt optimum and salt tolerance as either freshwater, brackish or marine. Except betaine, all organic osmotica known to occur in cyanobacteria, were found. The results showed no clear correlation between the chemical nature of the organic solute and the salt optimum or salt tolerance of the cyanobacteria examined, indicating that these solutes are not specific to this marine habitat. All strains belonging to the Nostoc/Anabea-group accumulated sucrose as the sole organic osmoticum. The marine, heterocystous Calothrix sp. accumulated trehalose. All strains of the LPP-group (Lyngbya, Plectonema, Phormidium) accumulated glucosylglycerol as sole or primary organic solute. Some LPP-strains accumulated a disaccharide as a secundary solute, e.g. sucrose or trehalose. Gloeocapsa, Synechocystis and Spirulina accumulated glucosylglycerol. Two marine Oscillatoria accumulated trehalose, whereas a freshwater Oscillatoria with a broad salinity tolerance, accumulated sucrose.Analysis of field samples of the microbial mats demonstrated the presence of glycerol, glucosylglycerol, sucrose and trehalose. The relative abundance of the different compounds was related to the species composition as could be predicted from laboratory observations. These data suggest that these carbohydrates have a function in maintaining osmotic balance in the organisms within the microbial mat.     

Volatile organic compounds of some Trichoderma spp. increase growth and induce salt tolerance in Arabidopsis thaliana

Many beneficial effects of Trichoderma spp. on plant growth and/or resistance to biotic/abiotic stresses can result from the production of bioactive compounds including volatile organic compounds (VOCs). We evaluated the effects of the volatile mixtures from 13 strains of different Trichoderma species on induction of tolerance to salt stress (100 mM NaCl) as well as growth promotion of Arabidopsis thaliana. Plants responded differently due to the presence of VOCs from various Trichoderma species ranging from both growth promotion and induction of salt tolerance to no significant changes under any of the conditions tested. In plants exposed for 2 weeks to VOCs of the selected strain, i.e. Trichoderma koningii, there was less H₂O₂ accumulation under salt stress compared to that in control plants. This result may reflect the possible role of VOCs of this strain in plant protection against oxidative damage under salt stress. Together, induction of salt tolerance using VOCs should be added to the known mechanisms of plant vigor enhancement by Trichoderma spp.     

Role of light and photosynthesis on the acclimation process of the cyanobacteriumSpirulina platensis to salinity stress

The response ofSpirulina platensis cells to salinity stress was studied. Once adapted to the higher osmoticum, photosynthetic parameters such as the maximum rate of photosynthesis under saturating irradiance (P_max) and the initial slope of the P-I curve () are reduced by 15% and 25% in 0.5 M NaCl grown cells, respectively. Salt-adapted cells have a modified biochemical composition; reduced protein and chlorophyll content, and an increased level of carbohydrates. The reduction in the photosynthetic capacity of the salt-adaptedSpirulina cells reflects a lower ability to utilize light energy and results in an increase in the susceptibility of the stressed cells to photoinhibition. This conclusion is supported by the finding that cultures exposed to salt stress show not only a decrease in growth rate (), but lose the ability to respond to increased irradiance with an increase in growth. The use of variable fluorescence as a fast and reliable measurement to follow the changes in PSII of salt-stressesSpirulina cells enables following the early events of salinity shock. It indicates that as soon as the cells are exposed to salt, a protection mechanism is induced. This mechanism does not require any protein synthesis and may take place even in the dark, though at somewhat reduced effectiveness. The significance of the result in providing a better understanding of the interaction between two environmental stresses — light and salinity — and their application in the outdoor mass cultivation ofSpirulina are discussed.Author for correspondence     

The inhibition of photosynthesis and photorespiration in isolated mesophyll cells of Phaseolus and Lycopersicon by reduced osmotic potentials

Mesophyll cells isolated from Phaseolus vulgaris and Lycopersicon esculentum show decreasing photosynthetic rates when suspended in media containing increasing concentrations of osmoticum. The photosynthetic activity was sensitive to small changes in osmotic potential over a range of sorbitol concentrations from 0.44 M (−1.08 MPa) to 0.77 M (−1.88 MPa). Photorespiration assayed by ¹⁴CO₂ release in CO₂-free air and by ¹⁴CO₂ release from the oxidation of [1–¹⁴C] glycolate also decreased as the osmotic potential of the incubation medium was reduced. The CO₂ compensation points of the cells increased with increasing concentration of osmoticum from approximately 60 μ I⁻¹1 at −1.08 MPa to 130 μl 1⁻¹ for cells stressed at −1.88 MPa. Changes in photosynthetic and photorespiratory activities occurred at moderate osmotic potentials in these cells suggesting that in whole leaves during a reduction in water potential, non- stomatal inhibition of CO₂ assimilation and glycolate pathway metabolism occurs simultaneously with stomatal closure.     

Sustained growth promotion in arabidopsis with long-term exposure to the beneficial soil bacterium Bacillus subtilis (GB03)

Volatile emissions from the commercial growth promoting soil bacterium Bacillus subtilis (GB03) are effective in augmenting short-term growth, photosynthetic capacity and salt tolerance in Petri-dish grown arabidopsis seedlings. In contrast, the impact sustained GB03 volatile exposure on plant growth and development has yet to be examined. here is provided physical and physiological data establishing that bacterial volatiles induce long-term growth promotion, elevated photosynthetic capacity and iron accumulation, as well as delayed albeit higher seed count compared with water-treated control plants. Plants were grown unrestricted in double Magenta boxes containing solid MS media for up to twelve weeks with GB03 volatiles introduced in separate containers within the chamber so that plant bacterial interactions were only by air-borne transmission. These results establish that GB03 volatiles induce sustained beneficial effects on Arabidopsis growth including robust and extended vegetative growth followed by elevated seed set.Key words: Bacillus subtilis (GB03), photosynthetic efficiency, plant growth promoting rhizobacteria (PGPR), seed set, volatile organic compound (VOCs)     

Physiological significance of proline and glycinebetaine: Maintaining photosynthesis during NaCl stress in wheat

Experiments on the physiological significance of accumulation of proline and glycinebetaine (GB) in sustaining photosynthesis during salt stress in wheat in vivo showed that pre-treatment with GB, but not proline, alleviated NaCl-induced stomatal and non-stomatal inhibition of photosynthesis completely. A permeating and non-dissociating osmoticum, 3-orthomethyl-glucopyranose, also alleviated NaCl-induced perturbations of photosynthesis, suggesting that GB may work by maintaining chloroplast volume and not by specific effects on photosynthetic processes.