Different tolerances and responses to low temperature and darkness between waterbloom forming cyanobacterium Microcystis and a green alga Scenedesmus

The dynamics of planktonic cyanobacteria in eutrophicated freshwaters play an important role in formation of annual summer blooms, yet overwintering mechanisms of these water bloom forming cyanobacteria remain unknown. The responses to darkness and low temperature of three strains (unicellular Microcystis aeruginosa FACHB-905, colonial M. aeruginosa FACHB-938, and a green alga Scenedesmus quadricauda FACHB-45) were investigated in the present study. After a 30-day incubation under darkness and low temperature, cell morphology, cell numbers, chlorophyll a, photosynthetic activity (ETRmax and I-k), and malodialdehyde (MDA) content exhibited significant changes in Scenedesmus. In contrast, Microcystis aeruginosa cells did not change markedly in morphology, chlorophyll a, photosynthetic activity, and MDA content. The stress caused by low temperature and darkness resulted in an increase of the antioxidative enzyme-catalase (CAT) in all three strains. When the three strains re-grew under routine cultivated condition subjected to darkness and low temperature, specific growth rate of Scenedesmus was lower than that of Microcystis. Flow cytometry (FCM) examination indicated that two distinct types of metabolic response to darkness and low temperature existed in the three strains. The results from the present study reveal that the cyanobacterium Microcystis, especially colonial Microcystis, has greater endurance and adaptation ability to the stress of darkness and low temperature than the green alga Scenedesmus.

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 sand burial on biomass, chlorophyll fluorescence and extracellular polysaccharides of man-made cyanobacterial crusts under experimental conditions

Soil cyanobacterial crusts occur throughout the world, especially in the semiarid and arid regions. It always encounters sand burial, which is an important feature of mobile sand dunes. A greenhouse 41 study was conducted to determine the effects of sand burial on biomass, chlorophyll fluorescence and extracellular polysaccharides of man-made cyanobacterial crusts in six periods of time (0, 5, 10, 15, 20 and 30 d after burying) and at five depths (0, 0.2, 0.5, 1 and 2cm). The results indicated that with the increase of the burial time and burial depth extracellular polysaccharides content and Fv/Fm decreased correspondingly and there were no significant differences between 20 and 30 burial days under different burial depths. The degradation of chlorophyll a content appeared only at 20 and 30 burial days and there was also no significant difference between them under different burial depths. It was also observed a simultaneous decrease of the values of the Fv/Fm and the content of extracellular polysaccharides happened in the crusted cyanobacterium Microcoleus vaginatus Gom. It may suggest that there exists a relationship between extracellular polysaccharides and recovery of the activity of photosystem II (PS II) after rehydration.

Effects of iron on growth, pigment content, photosystem II efficiency, and siderophores production of Microcystis aeruginosa and Microcystis wesenbergii

Changes in growth, photosynthetic pigments, and photosystem II (PS II) photochemical efficiency as well as production of siderophores of Microcystis aeruginosa and Microcystis wesenbergii were determined in this experiment. Results showed growths of M. aeruginosa and M. wesenbergii, measured by means of optical density at 665 nm, were severely inhibited under an iron-limited condition, whereas they thrived under an iron-replete condition. The contents of chlorophyll-a, carotenoid, phycocyanin, and allophycocyanin under an iron-limited condition were lower than those under an iron-replete condition, and they all reached maximal contents on day 4 under the iron-limited condition. PS II photochemical efficiencies (maximal PS II quantum yield), saturating light levels (I-k ) and maximal electron transport rates (ETRmax) of M. aeruginosa and M. wesenbergii declined sharply under the iron-limited condition. The PS II photochemical efficiency and ETRmax of M. aeruginosa rose , whereas in the strain of M. wesenbergii, they declined gradually under the iron-replete condition. In addition, I-k of M. aeruginosa and M. wesenbergii under the iron-replete condition did not change obviously. Siderophore production of M. aeruginosa was higher than that of M. wesenbergii under the iron-limited condition. It was concluded that M. aeruginosa requires higher iron concentration for physiological and biochemical processes compared with M. wesenbergii, but its tolerance against too high a concentration of iron is weaker than M. wesenbergii.

Responses of dark respiration in the light to desiccation and temperature in the intertidal macroalga, Ulva lactuca (Chorophyta) during emersion

Dark respiration (nonphotorespiratory mitochondrial CO2 release) in the light (R-L) of the intertidal macroalga Ulva lactuca (Chorophyta) during emersion was investigated with respect to its response to variations in temperature and desiccation. R-L was estimated by CO2 gas-exchange analysis using the Kok effect method, whereas dark respiration in darkness (R-D) was determined from CO2 release at zero light. Rates of R, were significantly and consistently lower than those of R-D in emersed U. lactuca across all the temperature and desiccation levels measured. This demonstrated that dark respiration was partially depressed in the light, with the percentage inhibition ranging from 32 to 62%. Desiccation exerted a negative effect on R-L and R-D at a high temperature, 33 degrees C, whereas it had much less effect on respiration at low and moderate temperatures, 23 and 28 degrees C. In general, R-L and R-D increased with increasing temperature in U. lactuca during all stages of emersion but responded less positively to temperature change with increasing desiccation. Additionally, the Q(10) value (i.e. the proportional increase of respiration for each 10 degrees C rise in temperature) for R-L calculated over the temperature range of 23 to 33 degrees C was significantly higher than that for R-D in U. lactuca during the initial stages of emersion. Respiratory carbon loss as a percentage of gross photosynthetic carbon gain increased with increasing temperature and/or desiccation but was significantly reduced when estimated using R-L rather than R-D. It is suggested that measurements of R-L and how it changes in a variable environment are as important as estimates of R-D and photosynthesis in determining simultaneous balance between photosynthetic carbon uptake and respiratory carbon loss and in modeling the net daily carbon gain for an intertidal macroalga.

Vertical mixing within the epilimnion modulates UVR-induced photoinhibition in tropical freshwater phytoplankton from southern China

1. The importance of vertical mixing in modulating the impact of UVR on phytoplankton photosynthesis was assessed in a tropical, shallow lake in southern China from late winter to mid-spring of 2005. 2. Daily cycles of fluorescence measurements (i.e. photosynthetic quantum yield, Y) were performed on both 'static' and in situ samples. Static samples were of surface water incubated at the surface of the lake under three radiation treatments - PAB (PAR + UVR, 280-700 nm), PA (PAR + UV-A, 320-700 nm) and P (PAR, 400-700 nm). In situ samples were collected every hour at three different depths - 0, 0.5 and 1 m. 3. The general daily pattern was of a significant decrease in Y from early morning towards noon, with partial recovery in the afternoon. Samples incubated under static conditions always had lower Y than those under in situ conditions at the same time of the day. 4. Under stratified conditions, no overall impact of UVR impact could be detected in situ when compared with the static samples. Further rapid vertical mixing not only counteracted the impact of UVR but also stimulated photosynthetic efficiency. 5. Based on these measurements of fluorescence, the mixing speed of cells moving within the epilimnion was estimated to range between 0.53 and 6.5 cm min(-1). 6. These data show that mixing is very important in modulating the photosynthetic response of phytoplankton exposed to natural radiation and, hence, strongly conditions the overall impact of UVR on aquatic ecosystems.

Effects of solar UV radiation on germination of conchospores and morphogenesis of sporelings in Porphyra haitanensis (Rhodophyta)

The effects of ultraviolet radiation (UVR 280-400 nm) on the germination of Porphyra haitanensis conchospores and on the growth and morphogenesis of the subsequent sporelings were investigated by culturing the released conchospores under natural sunlight from 29 September to 6 October 2005. Germination increased with time and was faster when UV-B was excluded using cut-off filters. There were significant negative effects of UV-B radiation on growth and cell division of sporelings, with decreases up to 18% for thallus length, between 6 and 18% for thallus width, up to 29% for thallus area, and between 6 and 14% for cell size as compared to PAR-controls. UV-A had a significant positive effect on morphogenesis, enhancing the formation of sporelings with cells dividing transversely; on the other hand, UV-B delayed the formation of such sporelings. We also tested the effects of solar UVR on the growth of P. haitanensis juveniles and found no significant effects. Our results indicate that UV-A has an important role in the germination and morphogenesis of the species, but on the other hand, sporelings of P. haitanensis are more sensitive to UV-B radiation than juveniles.

Solar UV radiation drives CO2 fixation in marine phytoplankton: A double-edged sword

Photosynthesis by phytoplankton cells in aquatic environments contributes to more than 40% of the global primary production (Behrenfeld et al., 2006). Within the euphotic zone (down to 1% of surface photosynthetically active radiation [PAR]), cells are exposed not only to PAR (400-700 nm) but also to UV radiation (UVR; 280-400 nm) that can penetrate to considerable depths (Hargreaves, 2003). In contrast to PAR, which is energizing to photosynthesis, UVR is usually regarded as a stressor (Hader, 2003) and suggested to affect CO2-concentrating mechanisms in phytoplankton (Beardall et al., 2002). Solar UVR is known to reduce photosynthetic rates (Steemann Nielsen, 1964; Helbling et al., 2003), and damage cellular components such as D1 proteins (Sass et al., 1997) and DNA molecules (Buma et al., 2003). It can also decrease the growth (Villafane et al., 2003) and alter the rate of nutrient uptake (Fauchot et al., 2000) and the fatty acid composition (Goes et al., 1994) of phytoplankton. Recently, it has been found that natural levels of UVR can alter the morphology of the cyanobacterium Arthrospira (Spirulina) platensis (Wu et al., 2005b). On the other hand, positive effects of UVR, especially of UV- A (315-400 nm), have also been reported. UV- A enhances carbon fixation of phytoplankton under reduced (Nilawati et al., 1997; Barbieri et al., 2002) or fast-fluctuating (Helbling et al., 2003) solar irradiance and allows photorepair of UV- B-induced DNA damage (Buma et al., 2003). Furthermore, the presence of UV-A resulted in higher biomass production of A. platensis as compared to that under PAR alone (Wu et al., 2005a). Energy of UVR absorbed by the diatom Pseudo-nitzschia multiseries was found to cause fluorescence (Orellana et al., 2004). In addition, fluorescent pigments in corals and their algal symbiont are known to absorb UVR and play positive roles for the symbiotic photosynthesis and photoprotection (Schlichter et al., 1986; Salih et al., 2000). However, despite the positive effects that solar UVR may have on aquatic photosynthetic organisms, there is no direct evidence to what extent and howUVR per se is utilized by phytoplankton. In addition, estimations of aquatic biological production have been carried out in incubations considering only PAR (i. e. using UV-opaque vials made of glass or polycarbonate; Donk et al., 2001) without UVR being considered (Hein and Sand-Jensen, 1997; Schippers and Lurling, 2004). Here, we have found that UVR can act as an additional source of energy for photosynthesis in tropical marine phytoplankton, though it occasionally causes photoinhibition at high PAR levels. While UVR is usually thought of as damaging, our results indicate that UVR can enhance primary production of phytoplankton. Therefore, oceanic carbon fixation estimates may be underestimated by a large percentage if UVR is not taken into account.

Response of Microcystis to copper stress - Do phenotypes of Microcystis make a difference in stress tolerance?

To elucidate the role of phenotype in stress-tolerant bloom-forming cyanobacterium Microcystis, two phenotypes of M. aeruginosa-unicellular and colonial strains were selected to investigate how they responded to copper stress. Flow cytometry (FCM) examination indicated that the percents of viable cells in unicellular and colonial Microcystis were 1.92-2.83% and 72.3-97.51%, respectively, under 0.25 mg l(-1) copper sulfate treatment for 24 h. Upon exposure to 0.25 mg l(-1) copper sulfate, the activities of antioxidative enzyme, such as superoxide dismutase (SOD) and catalase (CAT), were significantly increased in colonial Microcystis compared to unicellular Microcystis. Meanwhile, the values of the photosynthetic parameters (F-v/F-m, ETRmax and oxygen evolution rate) decreased more rapidly in unicellular Microcystis than in colonial Microcystis. The results indicate that colonial Microcystis has a higher endurance to copper than unicellular Microcystis. This suggests that the efficient treatment concentration of copper sulfate as algaecides will be dependent on the phenotypes of Microcystis. (C) 2006 Elsevier Ltd. All rights reserved.

Lysis of Aphanizomenon flos-aquae (Cyanobacterium) by a bacterium Bacillus cereus

A phytoplankton-lytic (PL) bacterium, Bacillus cereus, capable of lysing the bloom-forming cyanobacterium. Aphanizomenon flos-aquae was isolated from Lake Dianchi of Yunnan province, China. This bacterium showed lytic activities against a wide range of cyanobacteria/algae, including A. flos-aquae, Microcystis viridis, Microcystis wesenbergi, Microcystis aeruginosa, Chlorella ellipsoidea, Oscillatoria tenuis, Nostoc punctiforme, Anabaena flos-aquae, Spirulina maxima, and Selenastrum capricornutum. Chlorophyll a contents, phycocyanin contents, and photosynthetic activities of the A. flos-aquae decreased evidently in an infected culture for a period. Bacterium B. cereus attacked rapidly A. flos-aquae cells by cell-to-cell contact mechanism. It was shown that the lysis of A. flos-aquae began with the breach of the cyanobacterial cell wall, and the cyanobacterial cell appeared abnormal in the presence of the PL bacterium. Moreover, transmission electron microscope examinations revealed that a close contact between the bacterium and the cyanobacterium was necessary for lysis. Some slime extrusions produced from B. cereus assisted the bacterial cells to be in close association with and lyse the cyanobacterial cells. These findings suggested that this bacterium could play an important role in controlling the Aphanizomenon blooms in freshwaters. (c) 2006 Elsevier Inc. All rights reserved.

Effects of salt stress on carbohydrate metabolism in desert soil alga Microcoleus vaginatus Gom.

The effects of salt stress on carbohydrate metabolism in Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crusts, were investigated in the present study. Extracellular total carbohydrates and exopolysaccharides (EPS) in the culture medium produced by M. vaginatus increased significantly during the growth phase and reached a maximum during the stationary phase. The production of extracellular carbohydrates also significantly increased under higher salt concentrations, which was attributed to an increase in low molecular weight carbohydrates. In the presence of NaCl, the production of cellular total carbohydrates decreased and photosynthetic activity was impaired, whereas cellular reducing sugars, water-soluble sugars and sucrose content and sucrose phosphate synthase activity increased, reaching a maximum in the presence of 200 mmol/L NaCl. These parameters were restored to original levels when the algae were transferred to a non-saline medium. Sodium and K+ concentrations of stressed cells decreased significantly and H+-ATPase activity increased after the addition of exogenous sucrose or EPS. The results suggest that EPS and sucrose are synthesized to maintain the cellular osmotic equilibrium between the intra- and extracellular environment, thus protecting algal cells from osmotic damage, which was attributed to the selective exclusion of cellular Na+ and K+ by H+-ATPase.

Improving photosynthesis of microalgae by changing the ratio of light-harvesting pigments

Changing the ratio of light-harvesting pigments was regarded as an efficient way to improve the photosynthesis rate in microalgae, but the underlying mechanism is still unclear. In the present study, a mutant of Anabeana simensis (called SP) was selected from retrieved satellite cultures. Several parameters related with photosynthesis, such as the growth, photosynthesis rate, the content of photosynthetic pigment, low temperature fluorescence spectrum (77K) and electron transport rate, were compared with those of the wild type. It was found that the change in the ratio of light-harvesting pigments in the mutant led to more efficient light energy transfer and usage in mutant than in the wild type. This may be the reason why the mutant had higher photosynthesis and growth rates.

Ecophysiological characteristics of four intertidal marine macroalgae during emersion along Shantou coast of China, with a special reference to the relationship of photosynthesis and CO2

Intertidal marine macroalgae experience periodical exposures during low tide due to their zonational distribution. The duration of such emersion leads to different exposures of the plants to light and aerial CO2, which then affect the physiology of them to different extents. The ecophysiological responses to light and CO2 were investigated during emersion in two red algae Gloiopeltis furcata and Gigartina intermedia, and two brown algae Petalonia fascia and Sargassum hemiphyllum, growing along the Shantou coast of China. The light-saturated net photosynthesis in G. furcata and P. fascia showed an increase followed by slightly desiccation, whereas that in G. intermedia and S. hemiphyllum exhibited a continuous decrease with water loss. In addition, the upper-zonated G. furcata and P. fascia, exhibited higher photosynthetic tolerance to desiccation and required higher light level to saturate their photosynthesis than the lower-zonated G. intemedia and S. hemiphyllum. Desiccation had less effect on dark respiration in these four algae compared with photosynthesis. The light-saturated net photosynthesis increased with increased CO2 concentrations, being saturated at CO2 concentrations higher than the present atmospheric level in G. furcata, G. intermedia and S. hemiphyllum during emersion. It was evident that the relative enhancement of photosynthesis by elevated CO, in those three algae increased, though the absolute values of photosynthetic enhancement owing to CO2 increase were reduced when the desiccation statuses became more severe. However, in the case of desiccated P. fascia (water loss being greater than 20 %), light saturated net photosynthesis was saturated with current ambient atmospheric CO2 level. It is proposed that increasing atmospheric CO2 will enhance the daily photosynthetic production in intertidal macroalgae by varied extents that were related to the species and zonation.

Regulation of gamete release in the economic brown seaweed Hizikia fusiforme (Phaeophyta)

Gamete release is an essential event in artificial seeding of the economic brown seaweed, Hizikia fusiforme. Mass egg release occurred in the dark, with few eggs being discharged in the light. Release of eggs was elicited with eight practical salinity units (one PSU = 1 g sea salts l(-1)) and was inhibited by salinity levels > 32 PSU. Egg release was optimal at 23 degrees C, and was decreased by 72% in agitated seawater compared to unstirred seawater. Inhibitors of photosynthesis and ions channels suppressed egg release, indicating that this process was physiologically associated with photosynthetic activity and ion transport.

Positive effects of continuous low nitrate levels on growth and photosynthesis of Alexandrium tamarense (Gonyaulacales, Dinophyceae)

The growth and photosynthesis of Alexandrium tamarense (Lebour) Balech in different nutrient conditions were investigated. Low nitrate level (0.0882 mmol/L) resulted in the highest average growth rate from day 0 to day 10 (4.58 x 10(2) cells mL(-1) d(-1)), but the lowest cell yield (5420 cells mL(-1)) in three nitrate level cultures. High nitrate-grown cells showed lower levels of chlorophyll a-specific and cell-specific light-saturated photosynthetic rate (P-m(chl a) and P-m(cell)), dark respiration rate (R-d(chl a) and R-d(cell)) and chlorophyll a-specific apparent photosynthetic efficiency (alpha(chl a)) than was seen for low nitrate-grown cells; whereas the cells became light saturated at higher irradiance at low nitrate condition. When cultures at low nitrate were supplemented with nitrate at 0.7938 mmol/L in late exponential growth phase, or with nitrate at 0.7938 mmol/L and phosphate at 0.072 mmol/L in stationary growth phase, the cell yield was drastically enhanced, a 7-9 times increase compared with non-supplemented control culture, achieving 43 540 cells mL(-1) and 52 300 cells mL(-1), respectively; however, supplementation with nitrate in the stationary growth phase or with nitrate and phosphate in the late exponential growth phase increased the cell yield by no more than 2 times. The results suggested that continuous low level of nitrate with sufficient supply of phosphate may facilitate the growth of A. tamarense.