Draft genome sequence of Arthrospira platensis C1 (PCC9438)

Arthrospira platensis is a cyanobacterium that is extensively cultivated outdoors on a large commercial scale for consumption as a food for humans and animals. It can be grown in monoculture under highly alkaline conditions, making it attractive for industrial production. Here we describe the complete genome sequence of A. platensis C1 strain and its annotation. The A. platensis C1 genome contains 6,089,210 bp including 6,108 protein-coding genes and 45 RNA genes, and no plasmids. The genome information has been used for further comparative analysis, particularly of metabolic pathways, photosynthetic efficiency and barriers to gene transfer.     

Probing the mechanism of a cyanobacterial Δ9 fatty acid desaturase from Spirulina platensis C1 (Arthrospira sp. PCC 9438)

The initial and rate determining step in the mechanism of fatty acid desaturases has been proposed to be breakage of one of the C---H bonds at the site of the incipient double bond. This has been investigated and supported for a number of eukaryotic fatty acid desaturases through the use of kinetic isotope effect experiments with deuterated substrates. In order to probe the reaction catalyzed by the cyanobacterial Δ9 desaturase and compare it to the eukaryotic desaturases, the desC gene of Spirulina platensis, strain C1 (Arthrospira sp. PCC 9438) was expressed in a desaturase mutant of baker's yeast. Kinetic isotope effects were performed by culturing yeast transformants with deuterated thia-substituted stearic acids. A large kinetic isotope effect was found for the 9 position, in qualitative agreement with results from eukaryotic desaturases.     

Photosynthetic and respiratory electron transport in the alkaliphilic cyanobacterium Arthrospira (Spirulina) platensis

Photosynthetic and respiratory electron transport and their interplay with ion transport have been studied in Arthrospira platensis, a filamentous alkaliphilic cyanobacterium living in hypersaline lakes. As typical for alkaliphiles, A. platensis apparently does not maintain an outward positive pH gradient at its plasma membrane. Accordingly, sodium extrusion occurs via an ATP-dependent primary sodium pump, in contrast to the Na⁺/H⁺ antiport in most cyanobacteria. A. platensis is strongly dependent on sodium/bicarbonate symport for the uptake of inorganic carbon. Sodium extrusion in the presence of the Photosystem II inhibitor diuron indicates that a significant amount of ATP is supplied by cyclic electron transport around Photosystem I, the content of which in A. platensis is exceptionally high. Plastoquinol is oxidized by two parallel pathways, via the cytochrome b ₆ f complex and a putative cytochrome bd complex, both of which are active in the light and in the dark. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization of the iron-containing superoxide dismutase and its response to stress in cyanobacterium Spirulina (Arthrospira) platensis

Superoxide dismutase (SOD) is considered a primary antioxidant which defends against reactive oxygen species that are induced by environmental stress. In this study, we examined changes in SOD activity and expression in the cyanobacterium Spirulina (Arthrospira) platensis under iron and salinity stress; we characterized its induction under these stress conditions and we overexpressed the enzyme in a bacterial host for preliminary characterization. Analysis of SOD isoforms concludes that S. platensis was found to regulate only the iron-containing SOD isoform (FeSOD) in response to two types of stress that were tested. The FeSOD expression (on the level of both mRNA and enzyme activity) was induced by the stress conditions of salinity and iron levels. The FeSOD from S. platensis was overexpressed in Escherichia coli BL21. The recombinant FeSOD protein (about 23 kDa) was purified for characterization. It showed high specific activity and pH stability at 6.0–9.0, and it is relatively thermostable, retaining 45 % of its activity after 30 min at 90 °C. Phylogenetic analysis reveals that S. platensis FeSOD is grouped with the FeSODs from other cyanobacterial species and separated from those of the eukaryotic Chlorophyta, suggesting that the FeSOD gene may be used as a molecular marker in physiological, phylogenetic, and taxonomic studies. This study also suggests that the increased activity and expression of SOD may play a role in algal survival under stress conditions.     

Photosynthetically active and UV radiation act in an antagonistic way in regulating buoyancy of Arthrospira (Spirulina) platensis (cyanobacterium)

Buoyancy provided by gas vesicles has been suggested to play an important role in regulating vertical distribution and nutrient acquisition in cyanobacteria. However, little is known about how changes in UV radiation (UVR, 280–400 nm) would affect the buoyancy. We have shown here that the floatation activity of the economically important cyanobacterium Arthrospira platensis (D-0083) decreased with increased photosynthetic rates associated with increased photosynthetically active radiation (PAR), but it decreased less in the presence of UVR, which resulted in inhibitory effects. When the cells were grown under isoenergetic levels of solar PAR or UVR alone, they migrated downward under PAR but maintained buoyant under UVR. The buoyancy regulation of A. platensis depended on the exposed levels of PAR as well as UVR, which affected photosynthesis and growth in an antagonistic way. The buoyancy of A. platensis in water columns is likely to be dependant on diurnal photosynthetic performance regulated by solar radiation, and can hardly be considered as an active strategy to gain more energy during sunrise/sunset or to escape from harmful irradiation during the noon period.     

Molecular cloning and physical mapping of the nitrogenase structural genes from the filamentous, non-heterocystous cyanobacterium Pseudanabaena sp. PCC7409

Abstract Sequences homologous to the structural genes for dinitrogenase ( nifD and nifK ) and nitrogenase reductase ( nifH ) have been cloned from the filamentous, non-heterocystous cyanobacterium Pseudanabaena PCC7409. The nifHDK ⁻ homologous sequences were shown to reside on a 6.5-kb Eco RI restriction fragment by using a restriction fragment encoding the Klebsiella pneumoniae nifHDK genes as a heterologous hybridization probe. This 6.5-kb restriction fragment was cloned from a λ gt.wes Eco RI library of the Paseudanabaena sp. PCC7409 genome. This fragment was subcloned into the plasmid vector pUC9 to generate plasmid pPSU20. A detailed physical map of the insert in plasmid pPSU20 was determined, and relative positions of the nifH, nifD , and nifK homologous sequences on this fragment were determined by hybridization analysis with gene-specific fragments derived from the corresponding Anabaena sp. PCC7120 genes. The results indicate that these genes are contiguous in Pseudanabaena sp. PCC 7409 and are arranged in the order nifH, nifD , and nifK . This arrangement resembles that observed for other non-heterocystous cyanobacteria but differs from that observed for Anabaena, Calothrix , and Nostoc species.     

Effect of temperature and growth phase on fatty acid composition of the psychrophilic Vibrio sp. strain no. 5710

Abstract The cellular fatty acid composition of the psychrophilic Vibrio sp. strain No. 5710 isolated from a deep-sea sediment sample was analyzed. The presence of docosahexaenoic acid (22:6) was demonstrated as found previously in other deep-sea bacteria, and the relative amount of 22:6 decreased as the growth temperature increased. A temperature shift from 10°C to 0°C resulted in a relative increase of 22:6, and an opposite shift led to a decrease. In addition, hexadecanoic acid (16:0) was found to increase as the growth temperature increased. Therefore, it is suggested that the adaptation of 5710 to the growth temperature was carried out by the changes in the relative amounts of 22:6 and 16:0. When 5710 was grown at low temperature, it increased the relative amount of 22:6 presumably to maintain membrane fluidity at that temperature. In contrast, 5710 grown at high temperature probably maintained the membrane fluidity by increasing the amount of a saturated fatty acid, 16:0. Furthermore, observation of the fatty acid compositions at mid-exponential phase and early stationary phase revealed the proportions of several fatty acids, including a major fatty acid, 9- cis -hexadecenoic acid (16:1c, palmitoleic acid), were affected by the growth phase which may be due to the physiological difference between the growth phases.     

阳光紫外辐射和盐胁迫对钝顶螺旋藻光合作用和形态变化的耦合效应

将钝顶螺旋藻培养在含有不同NaCl浓度（0、0．4、0．8mol·L^-1）的培养基中,并置于室外全波段太阳辐射、阳光辐射滤除uvB以及光合有效辐射（PAR）三种辐射条件下,以探讨阳光uV辐射和盐胁迫对钝顶螺旋藻的耦合效应。结果表明,阳光uv辐射显著抑制钝顶螺旋藻的光化学效率,且随着盐浓度的提高,其受抑制程度加剧。D1蛋白含量在高水平PAR和uV辐射下都明显降低,而高盐浓度（0．8mol·L^-1NaCl）导致其含量进一步下降。此外,阳光uv辐射与盐胁迫的耦合作用使得藻丝发生明显断裂。     

低温胁迫对螺旋藻脱氢酶活性的影响

低温胁迫试验结果表明,处理的温度越低、天数越长,鄂尔多斯高原碱湖的钝顶螺旋藻S1、引进的钝顶螺旋藻S2和极大螺旋藻S3细胞内脱氢酶活性越低。在每一种处理下,脱氧酶活性高低的排列顺序均为S1〉S2〉S3。脱氢酶活性的Q10值变化范围排列为S1〈S2〈S3。S1的生理活动临界低温为5℃,S2和S3的为15℃。S1对低温有较强的适应性,为耐寒植物;S2和S3为冷敏感植物。     

Localization of NAD(P)H dehydrogenase in the cyanobacterium Synechocystis sp. strain PCC 6803

Antibodies raised against NdhH and NdhB detected these proteins in the thylakoid membrane of Synechocystis sp. strain PCC 6803, but not in a purified cytoplasmic membrane. We conclude that NAD(P)H dehydrogenase is largely, if not exclusively, confined to the thylakoid membrane.     

Effects of increased atmospheric CO2 and N supply on photosynthesis, growth and cell composition of the cyanobacterium Spirulina platensis (Arthrospira)

The consequences of the addition of CO2 (1%) in cultures of S. platensis are examined in terms of biomass yield, cell composition and external medium composition. CO2 enrichment was tested under nitrogen saturating and nitrogen limiting conditions. Increasing CO2 levels did not cause any change in maximum growth rate while it decreased maximum biomass yield. Protein and pigments were decreased and carbohydrate increased by high CO2, but the capability to store carbohydrates was saturated. C:N ratio remained unchanged while organic carbon released to the external medium was enhanced, suggesting that organic carbon release in S. platensis is an efficient mechanism for the maintenance of the metabolic integrity, balancing the cell C:N ratio in response to environmental CO2 changes. CO2 affected the pigment content: Phycocyanin, chlorophyll and carotenoids were reduced in around 50%, but the photosynthetic parameters were slightly changed. We propose that in S. platensis CO2 could act promoting degradation of pigments synthetised in excess in normal CO2 conditions, that are not necessary for light harvesting. Nitrogen assimilation was significantly not affected by CO2, and it is proposed that the inability to stimulate N assimilation by CO2 enrichment determined the lack of response in maximum growth rate. This revised version was published online in June 2006 with corrections to the Cover Date.     

Enhancement of proliferation and differentiation in bone marrow hematopoietic cells by Spirulina (Arthrospira) platensis in mice

This study evaluates whether Spirulina, including its components such as phycocyanin, enhances or sustains immune functions by promoting immune competent-cell proliferation or differentiation. The effects of Spirulina of a hot-water extract (SpHW), phycocyanin (Phyc), and cell-wall component extract (SpCW) on proliferation of bone marrow cells and induction of colony-forming activity in mice were investigated. The Spirulina extracts, SpHW, Phyc, and SpCW, enhanced proliferation of bone-marrow cells and induced colony-forming activity in the spleen-cell culture supernatant. Granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin-3 (IL-3) were detected in the culture supernatant of the spleen cells stimulated with the Spirulina extracts. Bone marrow-cell colony formation in soft-agar assay was also significantly induced by the blood samples and the culture supernatants of the spleen and Peyer's patch cells of the mice which ingested Spirulina extracts orally for 5 weeks in in vivo study. Ratios of neutrophils and lymphocytes in the peripheral blood and bone marrow, consequently, increased in the mice. Spirulina may have potential therapeutic benefits for improvement of weakened immune functions caused by, for example, the use of anticancer drugs.