Enhanced expression of antifreeze protein genes drives the development of freeze tolerance in an Antarctica isolate of Chlorella vulgaris

In adaptation to new environments, organisms may accumulate mutations within encoding sequences to modify protein characteristics or acquire mutations within regulatory sequences to alter gene expression levels. With the development of antifreeze capability as the example, this study presents the evidence that change in gene expression level is probably the most important mechanism for adaptive evolution in a green alga Chlorella vulgaris. C. vulgaris NJ-7, an isolate from Antarctica, possesses an 18S rRNA sequence identical to that of a temperate isolate, SAG211-11b/UTEX259, but shows much higher freeze tolerance than the later isolate. The chromosomal DNA/cDNA of four antifreeze genes, namely hiC6, hiC12, rpl10a and hsp70, from the two isolates of C. vulgaris were cloned and sequenced, and very few variations of deduced amino acid sequences were found. In contrast, the transcription of hiC6, hiC12 and rpl10a was greatly intensified in NJ-7 compared to that in UTEX259, which is correlated to the significantly enhanced freeze tolerance of the Antarctica isolate. (C) 2009 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

Effects of solar UV radiation on diurnal photosynthetic performance and growth of Gracilaria lemaneiformis (Rhodophyta)

Previous studies on diurnal photosynthesis of macroalgal species have shown that at similar levels of photosynthetically active radiation (PAR, 400-700nm) the photosynthetic rate is lower in the afternoon than in the morning. However, the impacts of solar ultraviolet radiation (UVR, 280-400nm) have been little considered. We investigated the diurnal photosynthetic behaviour of the economically significant red alga Gracilaria lemaneiformis in the absence or presence of UV-A+B or UV-B with a flow-through system. While UV-A and UV-B, respectively, inhibited noontime Pmax by 22% and 14% on the sunny days, UV-A during sunrise (PAR below about 50Wm-2) increased the net photosynthesis by about 8% when compared with PAR alone. UV-A + PAR also resulted in higher apparent photosynthetic efficiency in the morning than in the afternoon period than PAR alone. Nevertheless, integrated daytime photosynthetic production under solar PAR alone was higher than with either PAR + UV-A+B or PAR + UV-A. Relative growth rate in the long term (9 days) matched the integrated photosynthetic production in that UV-A led to 9-15% and UV-B to 19-22% reduction, respectively. UV-absorbing compounds were found to be higher in the thalli exposed to PAR+UV-A+B than under PAR alone, reflecting a protective response to UVR.

The accumulation and metabolism of astaxanthin in Scenedesmus obliquus (chlorophyceae)

The biosynthesis and metabolism of astaxanthin in coenobium alga Scenedesmus obliquus were investigated using a two-stage culture. The first stage was for the analysis of biosynthesis and accumulation of astaxanthin in alga cells which were cultured under induction conditions (incubation at 30 degrees C and illumination of 180 mu mol m(-2) s(-1)) for 48 h. The composition of the secondary carotenoids in algal cells was analyzed and seven ketocarotenoids were identified. The results implied that S. obliquus synthesized astaxanthin from beta-carotene through three possible pathways. In the second stage, the cultures were transferred to normal conditions (incubation at 25 C and illumination of 80 mu mol m(-2) s(-1)) for 72 h. Algal cells accumulated more chlorophyll and biosynthesis of secondary carotenoids terminated, the content of secondary carotenoids decreased from 59.48 to 6.57%. The results inferred that accumulation and metabolism of astaxanthin could be controlled by cultivated conditions which also could lead the mobilization of secondary carotenoids to support the algal cell growth. The results also implied that presumed conversions from astaxanthin to lutein or antheraxanthin could be modulated by culturing conditions. (C) 2008 Published by Elsevier Ltd.

Detection of the hepatotoxic microcystins in 36 kinds of cyanobacteria Spirulina food products in China

Gel filtration chromatography, ultra-filtration, and solid-phase extraction silica gel clean-up were evaluated for their ability to remove microcystins selectively from extracts of cyanobacteria Spirulina samples after using the reversed-phase octadecylsilyl ODS cartridge for subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The reversed-phase ODS cartridge/silica gel combination were effective and the optimal wash and elution conditions were: H2O (wash), 20% methanol in water (wash), and 90% methanol in water (elution) for the reversed-phase ODS cartridge, followed by 80% methanol in water elution in the silica gel cartridge. The presence of microcystins in 36 kinds of cyanobacteria Spirulina health food samples obtained from various retail outlets in China were detected by LC-MS/MS, and 34 samples (94%) contained microcystins ranging from 2 to 163 ng g(-1) (mean=1427 ng g(-1)), which were significantly lower than microcystins present in blue green alga products previously reported. MC-RR-which contains two molecules of arginine (R)-(in 94.4% samples) was the predominant microcystin, followed by MC-LR-where L is leucine-(30.6%) and MC-YR-where Y is tyrose-(27.8%). The possible potential health risks from chronic exposure to microcystins from contaminated cyanobacteria Spirulina health food should not be ignored, even if the toxin concentrations were low. The method presented herein is proposed to detect microcystins present in commercial cyanobacteria Spirulina samples.

Responses of photosynthetic activity in the drought-tolerant cyanobacterium, Nostoc flagelliforme to rehydration at different temperature

Photosynthetic activity during rehydration at four temperatures (5, 15, 25, 35 degrees C) was studied in a terrestrial, highly drought-tolerant cyanobacterium, Nostoc flagelliforme. At all the temperatures, the optimum quantum yield F-v/F-m increased rapidly within I It and then increased slowly during the process of rehydration. The increase in F-v/F-m at 25 and 35 degrees C was larger than that at 5 and 15 degrees C. In addition, the changes of initial intensity of fluorescence (F-0) and variable fluorescence (F-v) were more significant at 25 and 35 degrees C than those at 5 and 15 degrees C. Chlorophyll a content increased with the increase of temperature during the course of rehydration, with this being more pronounced at 25 and 35 degrees C. The photosynthetic rates at 25 and 35 degrees C were higher than those at 5 and 15 degrees C. Induction of chlorophyll fluorescence with sustained rewetting at 5 and 15 degrees C had two phases of transformation, whereas at 25 and 35 degrees C it had a third peak kinetic phase and showed typical chlorophyll fluorescence steps on rewetting for 24 h, representing a normal physiological state. A comparison of the chlorophyll fluorescence parameters, chlorophyll a content, and the chlorophyll fluorescence induction led to the conclusion that N. flagelliforme had a more rapid and complete recovery at 25 and 35 degrees C than that at 5 and 15 degrees C, although it could recover its photosynthetic activity at any of the four temperatures. (c) 2007 Published by Elsevier Ltd.

Physiological and biochemical responses of Scytonema javanicum (cyanobacterium) to salt stress

Scytonema javanicum (Kutz.) Born et Flah (cyanobacterium) is one of the species distributed widely in the crust of desert soils regularly subjected to severe water stress. To investigate the response of the species to salt stress, many physiological and biochemical parameters, including growth rate, ratio of variable fluorescence to maximum fluorescence (Fv/Fm), reactive oxidative species (ROS), malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD), were determined in culture. The results showed that 50 mM NaCl inhibited growth and Fv/Fm in the medium BG-110, and that the inhibition was maximum after 1-2 days' exposure to salt stress; 50 mM NaCl also increased the contents of ROS and MDA in treated cells, which suggests that salt stress may lead to oxidative damage and lipid peroxidation in the alga. Further, changes in the antioxidative enzymes SOD and CAT in the treated alga were consistent with changes in ROS and MDA at certain extent. These observations suggest that oxidative stress resulting from salt stress in S. javanicum could result in the production of antioxidative enzymes to counteract the oxidative damage, and the enzymes may contribute to the ability of S. javanicum to survive the adverse desert environment. (c) 2007 Elsevier Ltd. All rights reserved.

Effects of solar UV radiation on morphology and photosynthesis of filamentous cyanobacterium Arthrospira platensis

To study the impact of solar UV radiation (UVR) (280 to 400 nm) on the filamentous cyanobacterium Arthrospira (Spirulina) platensis, we examined the morphological changes and photosynthetic performance using an indoor-grown strain (which had not been exposed to sunlight for decades) and an outdoor-grown strain (which had been grown under sunlight for decades) while they were cultured with three solar radiation treatments: PAB (photosynthetically active radiation [PAR] plus UVR; 280 to 700 nm), PA (PAR plus UV-A; 320 to 700 nm), and P (PAR only; 400 to 700 nm). Solar UVR broke the spiral filaments of A. platensis exposed to full solar radiation in short-term low-cell-density cultures. This breakage was observed after 2 h for the indoor strain but after 4 to 6 h for the outdoor strain. Filament breakage also occurred in the cultures exposed to PAR alone; however, the extent of breakage was less than that observed for filaments exposed to full solar radiation. The spiral filaments broke and compressed when high-cell-density cultures were exposed to full solar radiation during long-term experiments. When UV-B was screened off, the filaments initially broke, but they elongated and became loosely arranged later (i.e., there were fewer spirals per unit of filament length). When UVR was filtered out, the spiral structure hardly broke or became looser. Photosynthetic 0, evolution in the presence of UVR was significantly suppressed in the indoor strain compared to the outdoor strain. UVR-induced inhibition increased with exposure time, and it was significantly lower in the outdoor strain. The concentration of UV-absorbing compounds was low in both strains, and there was no significant change in the amount regardless of the radiation treatment, suggesting that these compounds were not effectively used as protection against solar UVR. Self-shading, on the other hand, produced by compression of the spirals over adaptive time scales, seems to play an important role in protecting this species against deleterious UVR. Our findings suggest that the increase in UV-B irradiance due to ozone depletion not only might affect photosynthesis but also might alter the morphological development of filamentous cyanobacteria during acclimation or over adaptive time scales.

Present status and changes of the phytoplankton community after invasion of Neosalanx taihuensis since 1982 in a deep oligotrophic plateau lake, Lake Fuxian in the subtropical China

Phytoplankton assemblages in the subtrophical oligotrophic Lake Fuxian, the second deepest lake in China, were investigated monthly from September 2002 to August 2003. A total of 113 species belonging to seven phyla were identified, among them, a filamentous green alga, Mougeotia sp., dominated almost throughout the study period and comprised most of the total phytoplankton biomass. Mougeotia sp. has made a substantial development during the past decades: it was absent in 1957, only occasionally present in 1983, increased substantially in 1993, and became predominant in 2002-2003. It is likely that natural invasion of the Taihu Lake noodlefish (Neosalanx taihuensis) has led to a change of dominant herbivorous zooplankton from small to large calanoid, which has increased grazing pressure on small edible algae, and thus has indirectly favored the development of the inedible filamentous Mougeotia sp.

The rotifer assemblage in a shallow, eutrophic Chinese lake and its relationships with cyanobacterial blooms and crustacean zooplankton

Rotifer assemblage in the subtropical eutrophic Lake Chaohu was investigated monthly from September 2002 to August 2003. Forty-nine species belonging to 18 genera and 14 families were recorded. The highest densities of rotifer were observed during summer when there were heavy cyanobacterial blooms. There was a significant positive correlation between total rotifer density and the biomass of cyanobacteria. However, no correlations were found between the densities of rotifer and crustacean zooplankton, possibly owing to the paucity of large-bodied planktonic crustaceans. It is likely that the occurrence of cyanobacterial blooms not only caused the shift of dominant crustacean zooplankton from large species to smaller ones but also weakened the negative interaction between crustaceans and rotifers.

Comparative mechanisms of photosynthetic carbon acquisition in Hizikia fusiforme under submersed and emersed conditions

The economic seaweed Hizikia fusiforme (Harv.) Okamura (Sargassaceae, Phaeophyta) usually experiences periodical exposures to air at low tide. Photosynthetic carbon acquisition mechanisms were comparatively studied under submersed and emersed conditions in order to establish a general understanding of its photosynthetic characteristics associated with tidal cycles. When submersed in seawater, H fusiforme was capable of acquiring HCO3- as a source of inorganic carbon (Ci) to drive photosynthesis, while emersed and exposed to air, it used atmospheric CO2 for photosynthesis. The pH changes surrounding the H fusiforme fronds had less influence on the photosynthetic rates under emersed condition than under submersed condition. When the pH was as high as 10.0, emersed H fusiforme could photosynthesize efficiently, but the submersed alga exhibited very poor photosynthesis. Extracellular carbonic anhydrase (CA) played an important role in the photosynthetic acquisitions of exogenous Ci in water as well as in air. Both the concentrations of dissolved inorganic carbon in general seawater and CO2 in air were demonstrated to limit the photosynthesis of H fusiforme, which was sensitive to O-2. It appeared that the exogenous carbon acquisition system, being dependent of external CA activity, operates in a way not enough to raise intracellular CO2 level to prevent photorespiration. The inability of H fusiforme to achieve its maximum photosynthetic rate at the current ambient Ci levels under both submersed and emersed conditions suggested that the yield of aquaculture for this economic species would respond profitably to future increases in CO2 concentration in the sea and air.

Expression and molecular analysis of phbB gene in Chlamydomonas reinhardtii

The expression vector containing phbB and ble genes was constructed and transformed into cell-wall-deficient strain Chlamydomonas reinhardtii CC-849 by the glass-head method. The transgenic alga was selected and maintained in the TAP agar plates containing 10 mug/mL Zeomycin. Transgenic alga, which could express phbB at the transcriptional level, was obtained and further confirmed with PCR, Southern blot and RT-PCR-DNA hybridization analysis.

The vertical microdistribution of cyanobacteria and green algae within desert crusts and the development of the algal crusts

Substantial amounts of algal crusts were collected from five different desert experimental sites aged 42, 34, 17, 8 and 4 years, respectively, at Shapotou ( China) and analyzed at a 0.1 mm microscale of depth. It was found that the vertical distribution of cyanobacteria and microalgae in the crusts was distinctly laminated into an inorganic-layer (ca. 0.00 - 0.02 mm, with few algae), an algae-dense-layer ( ca. 0.02 - 1.0 mm) and an algae-sparse-layer ( ca. 1.0 - 5.0 mm). It was interesting to note that in all crusts Scytonema javanicum Born et Flah ( or Nostoc sp., cyanobacterium), Desmococcus olivaceus (Pers ex Ach., green alga) Laundon and Microcoleus vaginatus Gom. ( cyanobacterium) dominated at the depth of 0.02 - 0.05, 0.05 - 0.1 and 0.1 - 1.0 mm, respectively, from the surface. Phormidium tenue Gom. ( or Lyngbya cryptovaginatus Schk., cyanobacterium) and Navicula cryptocephala Kutz.( or Hantzschia amphioxys (Ehr.) Grun. and N. cryptocephala together, diatom) dominated at the depth of 1.0 - 3.0 and 3.5 - 4.0 mm, respectively, of the crusts from the 42 and 34 year old sites. It was apparent that in more developed crusts there were more green algae and the niches of Nostoc sp., Chlorella vulgaris Beij., M. vaginatus, N. cryptocephala and fungi were nearer to the surface. If lichens and mosses accounted for less than 41.5% of the crust surface, algal biovolume was bigger when the crust was older, but the opposite was true when the cryptogams other than algae covered more than 70%. In addition to detailed species composition and biovolume, analyses of soil physicochemical properties, micromorphologies and mineral components were also performed. It was found that the concentration of organic matter and nutrients, electric conductivity, silt, clay, secondary minerals were higher and there were more micro-beddings in the older crusts than the less developed ones. Possible mechanisms for the algal vertical microdistribtion at different stages and the impact of soil topography on crust development are discussed. It is concluded that biomethods ( such as fine species distribution and biovolume) were more precise than mineralogical approaches in judging algal crust development and thus could be a better means to measure the potentiality of algal crusts in desert amelioration.

Extracellular carbohydrate polymers from five desert soil algae with different cohesion in the stabilization of fine sand grain

Extracellular polymeric substances (EPS) from four filamentous cyanobacteria Microcoleus vaginatus, Scytonema javanicum, Phormidium tenue and Nostoc sp. and a coccoid single-cell green alga Desmococcus olivaceus that had been separated from desert algal crusts of Tegger desert of China, were investigated for their chemical composition, structure,and physical properties. The EPS contained 7.5-50.3% protein (in polymers ranging from 14 to more than 200 kD, SDS-PAGE) and 16.2-46.5% carbohydrate (110-460 kD, GFC). 6-12 kinds of monosaccharides, including 2-O-methyl rhamnose, 2-O-methyl glucose, and N-acetyl glucosamine were found. The main carbohydrate chains from M. vaginatus and S. javanicum consisted mainly of equal proportion of Man, Gal and Glc, that from P. tenue consisted mainly of arabinose, glucose and rhamnose. Arabinose was present in pyranose form, mainly alpha-L 1 --> 3 linked, with branches on C4 of almost half of the units. Glucose was responsible for the terminal units, in addition of having some units as beta1 --> 3 and some as beta1 --> 4 linked. Rhamnose was mainly 1 --> 3 linked with branches on C2 on half of the units. The carbohydrate polymer from D. olivaceus was composed mainly of beta1 --> 4 linked xylose, galactose and glucose. The galactose part was present both in beta-pyranose and -furanose forms. Arabinose in alpha-L-furanose form was mainly present as 1 --> 2 and 1 --> 2, 5 linked units, rhamnose only as alpha 1 --> 3 and xylose as beta 1 --> 4. The backbone of the polysaccharide from Nostoc sp. was composed of beta-1 --> 4 linked xylose, galactose and glucose. Most of the glucose was branched on position C6, terminal glucose and 2-O-methyl glucose units are also present. The relationship between structure, physical properties and potential biological function is discussed. (C) 2003 Elsevier Ltd. All rights reserved.