Allelopathic inhibition of phytoplankton by exudates from Stratiotes aloides

The allelopathic potential of exudates from the aquatic macrophyte Stratiotes aloides on the growth of phytoplankton was investigated. A selection of phytoplankton species, occurring in habitats similar to that of Stratiotes, was used: two cyanobacterial strains (toxic and non-toxic Microcystis aeruginosa), one green alga (Scenedesmus obliquus) and one eustigmatophyte (Nannochloropsis limnetica). The results indicate allelopathic effects of Stratiotes on phytoplankton in six of the eight cases, expressed in an extended duration of the initial biovolume doubling time. The overall inhibitory effect (8–51%) was strain-specific for the two cyanobacteria. We also studied the effect of irradiance on the allelopathic potential of exudates from Stratiotes. Irradiance influenced the response of Scenedesmus only. The inhibitory effect of Stratiotes exudates on the growth of this green alga was stronger at 35 μmol m⁻² s⁻¹ than at 105 μmol m⁻² s⁻¹. We conclude that Stratiotes has allelopathic effects on phytoplankton, and that irradiance can, but does not always determine the extent of the allelopathic inhibition. In our experiments, the sensitivity of cyanobacteria to Stratiotes exudates was not higher than for other phytoplankton strains, but within cyanobacteria, the toxic strain was more sensitive than the non-toxic one.     

Toxic and non-toxic strains of Microcystis aeruginosa induce temperature dependent allelopathy toward growth and photosynthesis of Chlorella vulgaris

Global warming was believed to accelerate the expansion of cyanobacterial blooms. However, the impact of changes due to the allelopathic effects of cyanobacterial blooms with or without algal toxin production on the ecophysiology of its coexisting phytoplankton species arising from global warming were unknown until recently. In this study, the allelopathic effects of toxic and non-toxic Microcystis aeruginosa strains on the growth of green alga Chlorella vulgaris and photosynthesis of the co-cultivations of C. vulgaris and toxic M. aeruginosa FACHB-905 or non-toxic M. aeruginosa FACHB-469 were investigated at different temperatures. The growth of C. vulgaris, co-cultured with the toxic or non-toxic M. aeruginosa strains, was promoted at 20 °C but inhibited at temperatures ≥25 °C. The inhibitory effects of the toxic and non-toxic M. aeruginosa strains on of the co-cultivations (C. vulgaris and non-toxic M. aeruginosa FACHB-469 or toxic M. aeruginosa FACHB-905) also linearly increased with elevated temperatures. Furthermore, toxic M. aeruginosa FACHB-905 induced more inhibition toward growth of C. vulgaris or P_max and R_d of the mixtures than non-toxic M. aeruginosa FACHB-469. C. vulgaris dominated over non-toxic M. aeruginosa FACHB-469 but toxic M. aeruginosa FACHB-905 overcame C. vulgaris when they were co-cultured in mesocosms in water temperatures from 20 to 25 °C. The results indicate that allelopathic effects of M. aeruginosa strains on C. vulgaris are both temperature- and species-dependent: it was stimulative for C. vulgaris at low temperatures such as 20 °C, but inhibitory at high temperatures (≥25 °C); the toxic strain was determined to be more harmful to C. vulgaris than the non-toxic one. This suggests that global warming may aggravate the ecological risk of cyanobacteria blooms, especially those with toxic species as the main contributors.     

Interspecific allelopathy in cyanobacteria: Cylindrospermopsin and Cylindrospermopsis raciborskii effect on the growth and metabolism of Microcystis aeruginosa

The biological role of cyanobacteria secondary metabolites is relatively unknown although several possible hypotheses have been discussed. In the following study the effect of cylindrospermopsin (CYN) and metabolites of non-CYN producing Cylindrospermopsis raciborskii strain on growth, alkaline phosphatase (ALP) activity and microcystin-LR (MC-LR) production in Microcystis aeruginosa was evaluated. Higher concentrations of CYN (10 and 50 μg L⁻¹) induced toxicity effects demonstrated by significant growth inhibition and M. aeruginosa cell necrosis. Lower concentrations of CYN (1 and 5 μg L⁻¹) slightly decreased growth rates but significantly up-regulated ALP activity. Moreover, under all studied CYN concentrations MC-LR production strongly decreased. Spent C. raciborskii medium mimicked the CYN action by inducing strong inhibition of M. aeruginosa growth and MC-LR production and through up-regulation of ALP activity. On the other hand, spent M. aeruginosa medium did not affect C. raciborskii growth and no alterations in ALP activity were observed. Co-culturing of these two species resulted in an increase of C. raciborskii contribution at the expense of M. aeruginosa. From the results we conclude that CYN can be involved in interspecific competition in cyanobacteria and that non-CYN producing C. raciborskii strains may produce a hitherto unknown bioactive compound(s) which can mimic CYN action.     

Mathematical modeling of Microcystis aeruginosa growth and [D-Leu1] microcystin-LR production in culture media at different temperatures

The effect of temperature (26 °C, 28 °C, 30 °C and 35 °C) on the growth of native CAAT-3-2005 Microcystis aeruginosa and the production of Chlorophyll-a (Chl-a) and Microcystin-LR (MC-LR) were examined through laboratory studies. Kinetic parameters such as specific growth rate (μ), lag phase duration (LPD) and maximum population density (MPD) were determined by fitting the modified Gompertz equation to the M. aeruginosa strain cell count (cells mL⁻¹). A 4.8-fold increase in μ values and a 10.8-fold decrease in the LPD values were found for M. aeruginosa growth when the temperature changed from 15 °C to 35 °C. The activation energy of the specific growth rate (Eμ) and of the adaptation rate (E₁/LPD) were significantly correlated (R² = 0.86). The cardinal temperatures estimated by the modified Ratkowsky model were minimum temperature = 8.58 ± 2.34 °C, maximum temperature = 45.04 ± 1.35 °C and optimum temperature = 33.39 ± 0.55 °C.Maximum MC-LR production decreased 9.5-fold when the temperature was increased from 26 °C to 35 °C. The maximum production values were obtained at 26° C and the maximum depletion rate of intracellular MC-LR was observed at 30–35 °C. The MC-LR cell quota was higher at 26 and 28 °C (83 and 80 fg cell⁻¹, respectively) and the MC-LR Chl-a quota was similar at all the different temperatures (0.5–1.5 fg ng⁻¹).The Gompertz equation and dynamic model were found to be the most appropriate approaches to calculate M. aeruginosa growth and production of MC-LR, respectively. Given that toxin production decreased with increasing temperatures but growth increased, this study demonstrates that growth and toxin production processes are uncoupled in M. aeruginosa. These data and models may be useful to predict M. aeruginosa bloom formation in the environment.     

Growth of eight Gambierdiscus (Dinophyceae) species: Effects of temperature,salinity and irradiance

Little is known about how the growth of individual Gambierdiscus species responds to environmental factors. This study examined the effects of temperature (15–34 °C), salinity (15–41) and irradiance (2–664 μmol photons m⁻² s⁻¹) on growth of Gambierdiscus: G. australes, G. belizeanus, G. caribaeus, G. carolinianus, G. carpenteri, G. pacificus and G. ruetzleri and one putative new species, Gambierdiscus ribotype 2. Depending on species, temperatures where maximum growth occurred varied between 26.5 and 31.1 °C. The upper and lower thermal limits for all species were between 31–34 °C and 15–21 °C, respectively. The shapes of the temperature vs. growth curves indicated that even small differences of 1–2 °C notably affected growth potentials. Salinities where maximum growth occurred varied between 24.7 and 35, while the lowest salinities supporting growth ranged from <14 to 20.9. These data indicated that Gambierdiscus species are more tolerant of lower salinities than is generally appreciated. Growth of all species began to decline markedly as salinities exceed 35.1–39.4. The highest salinity tested in this study (41), however, was lethal to only one species, Gambierdiscus ribotype 2. The combined salinity data indicated that differences in salinity regimes may affect relative species abundances and distributions, particularly when salinities are <20 and >35. All eight Gambierdiscus species were adapted to relatively low light conditions, exhibiting growth maxima at 50–230 μmol photons m⁻² s⁻¹ and requiring only 6–17 μmol photons m⁻² s⁻¹ to maintain growth. These low light requirements indicate that Gambierdiscus growth can occur up to 150 m depth in tropical waters, with optimal light regimes often extending to 75 m. The combined temperature, salinity and light requirements of Gambierdiscus can be used to define latitudinal ranges and species-specific habitats, as well as to inform predictive models.     

Nitrate and phosphate uptake kinetics of the harmful diatom Coscinodiscus wailesii,a causative organism in the bleaching of aquacultured Porphyra thalli

The large diatom Coscinodiscus wailesii is one of the problematic species which indirectly cause bleaching damage to “Nori” (Porphyra thalli) cultivation through competitive utilization of nutrients during its bloom. In the present study, we experimentally investigated the nitrate (N) and phosphate (P) uptake kinetics of C. wailesii, Harima-Nada strain. Maximum uptake rates (ρ_max), obtained by short-term experiments, were 58.3 and 95.5 pmol cell⁻¹ h⁻¹ for nitrate and 41.9 and 59.1 pmol cell⁻¹ h⁻¹ for phosphate at 9 and 20 °C, respectively. The half saturation constants for uptake (K_s) were 2.91 and 5.08 μM N and 5.62 and 6.67 μM P at 9 and 20 °C, respectively. The ρ_max values of C. wailesii, much higher than those of other marine phytoplankton species, suggest that C. wailesii is able to take up large amounts of nutrients from the water column. On the other hand, V_max/K_s (V_max; V_max = ρ_max/Q₀, Q₀; minimum cell quota) values of C. wailesii, which is a better measure to evaluate the competitive ability for nutrient uptake, were low in dominant diatom species. This parameter indicates that C. wailesii is disadvantaged compared to other diatom species in competing for nutrients, and the decreasing nutrient concentrations from winter to spring is an important factor limiting C. wailesii blooming in early spring.     

A comparative study of the use of inorganic carbon resources by Chara aspera and Potamogeton pectinatus

We studied the potential role of dissolved inorganic carbon (DIC) in determining vegetation dominance of Potamogeton pectinatus L. and Chara aspera Deth. ex Willd. by monitoring the seasonal dynamics of DIC in a shallow lake and comparing the use of DIC of the two species. The HCO₃⁻-concentration in summer dropped from 2.5 to <0.5 mM with seasonally increasing Chara biomass, whereas outside the vegetation concentrations remained at 2.5 mM. Inside Potamogeton spp. vegetation DIC decreased from 2.5 to ca. 0.75 mM HCO₃⁻. A growth experiment showed ash-free biomass for P. pectinatus was nearly two times as high as for C. aspera at 3 mM HCO₃⁻, but almost two times lower at 0.5 mM than at 3.0. In a separate experiment, P. pectinatus precultured at a relatively low HCO₃⁻-level had a lower net photosynthetic rate (P_max, 0.1 mmol O₂ g⁻¹ DW h⁻¹) than C. aspera (P_max, 0.1 mmol O₂ g⁻¹ DW h⁻¹) over the range of HCO₃⁻-concentrations tested (P_max, 0.14 mmol O₂ g⁻¹ DW h⁻¹). In response to CO₂ no significant differences between the compensation points (P. pectinatus, 28 mM; C. aspera 66 mM), were observed, but the photosynthetic rate increased faster than for C. aspera than for P. pectinatus. Under field conditions, the use of CO₂ is not important since inside vegetation CO₂-concentrations were below 10 μM, and thus, not available for photosynthesis of either species during the main part of the growth season. It is suggested that C. aspera may be a better competitor for HCO₃⁻ than P. pectinatus in conditions with a low HCO₃⁻ supply. As HCO₃⁻ is a strong limiting factor for growth inside the vegetation and probably the only carbon source available, the superior ability of C. aspera to use HCO₃⁻ may be an important factor explaining its present dominance in Veluwemeer.     

Growth and loss of distal tissue in blades of Lessonia nigrescens and Lessonia trabeculata (Laminariales)

Meristematic growth and loss of distal tissue from blades of two ecologically important species in the south-east Pacific, Lessonia nigrescens and Lessonia trabeculata, was evaluated during 1 year. Comparative growth was determined by a hole-punch method, loss of distal tissue from the blades was determined by subtracting final blade length (with loss) from expected blade lengths (without loss); growth and tissue loss were transformed to fresh biomass units for calculation of inter-algae differences. The results showed that blade elongation rate increased at the beginning of spring, and declined towards the end of summer, with mean values between 0.40 and 0.08 cm day⁻¹ for L. nigrescens, and 0.65–0.17 cm day⁻¹ for L. trabeculata. Loss of distal tissue varied seasonally when examined as length units for both species; with mean values between 0.24 and 0.10 cm day⁻¹ for L. nigrescens, and 0.51–0.25 cm day⁻¹ for L. trabeculata. Variations in fresh biomass units were only observed in Lessonia trabeculata, increasing in spring, with mean values to 0.13 g (fresh weight) day⁻¹. Annual growth and loss of distal tissue were higher in L. trabeculata (0.41 and 0.39 cm day⁻¹, respectively) than in L nigrescens (0.19 and 0.15 cm day⁻¹). When growth and tissue loss were considered as fresh biomass, monthly gains significantly outweighed loss of distal tissue in both species, but parallel results based on length data followed a different trend. L. trabeculata released about 50% of its growth biomass as particulate organic matter, while the comparative value for L. nigrescens was about 20%.     

Newly discovered role of the heterotrophic nanoflagellate Katablepharis japonica,a predator of toxic or harmful dinoflagellates and raphidophytes

Heterotrophic nanoflagellates are ubiquitous and known to be major predators of bacteria. The feeding of free-living heterotrophic nanoflagellates on phytoplankton is poorly understood, although these two components usually co-exist. To investigate the feeding and ecological roles of major heterotrophic nanoflagellates Katablepharis spp., the feeding ability of Katablepharis japonica on bacteria and phytoplankton species and the type of the prey that K. japonica can feed on were explored. Furthermore, the growth and ingestion rates of K. japonica on the dinoflagellate Akashiwo sanguinea—a suitable algal prey item—heterotrophic bacteria, and the cyanobacteria Synechococcus sp., as a function of prey concentration were determined. Among the prey tested, K. japonica ingested heterotrophic bacteria, Synechococcus sp., the prasinophyte Pyramimonas sp., the cryptophytes Rhodomonas salina and Teleaulax sp., the raphidophytes Heterosigma akashiwo and Chattonella ovata, the dinoflagellates Heterocapsa rotundata, Amphidinium carterae, Prorocentrum donghaiense, Alexandrium minutum, Cochlodinium polykrikoides, Gymnodinium catenatum, A. sanguinea, Coolia malayensis, and the ciliate Mesodinium rubrum, however, it did not feed on the dinoflagellates Alexandrium catenella, Gambierdiscus caribaeus, Heterocapsa triquetra, Lingulodinium polyedra, Prorocentrum cordatum, P. micans, and Scrippsiella acuminata and the diatom Skeletonema costatum. Many K. japonica cells attacked and ingested a prey cell together after pecking and rupturing the surface of the prey cell and then uptaking the materials that emerged from the ruptured cell surface. Cells of A. sanguinea supported positive growth of K. japonica, but neither heterotrophic bacteria nor Synechococcus sp. supported growth. The maximum specific growth rate of K. japonica on A. sanguinea was 1.01 d⁻¹. In addition, the maximum ingestion rate of K. japonica for A. sanguinea was 0.13 ng C predator⁻¹d⁻¹ (0.06 cells predator⁻¹d⁻¹). The maximum ingestion rate of K. japonica for heterotrophic bacteria was 0.019 ng C predator⁻¹d⁻¹ (266 bacteria predator⁻¹d⁻¹), and the highest ingestion rate of K. japonica for Synechococcus sp. at the given prey concentrations of up to ca. 10⁷ cells ml⁻¹ was 0.01 ng C predator⁻¹d⁻¹ (48 Synechococcus predator⁻¹d⁻¹). The maximum daily carbon acquisition from A. sanguinea, heterotrophic bacteria, and Synechococcus sp. were 307, 43, and 22%, respectively, of the body carbon of the predator. Thus, low ingestion rates of K. japonica on heterotrophic bacteria and Synechococcus sp. may be responsible for the lack of growth. The results of the present study clearly show that K. japonica is a predator of diverse phytoplankton, including toxic or harmful algae, and may also affect the dynamics of red tides caused by these prey species.     

Morphology and bloom dynamics of Cochlodinium geminatum (Schütt) Schütt in the Pearl River Estuary,South China Sea

An unarmored dinoflagellate bloom of Cochlodinium geminatum (Schütt) Schütt has been identified in the Pearl River Estuary, South China Sea during the severe dry season, from late October to early November, 2009, when temperature and salinity ranged between 20.0–27.2 °C and 10.6–33.4, respectively. Light and scanning electron microscopy were used to identify the characteristics of C. geminatum and provided the clear morphological structure for this species. The organism was primarily found in chains of two cells or single cell, and no longer chains were observed. Cells were irregularly spherical or slightly dorso-ventrally, with size ranged between 28 and 36 μm and longer than wide. A large nucleus in the center with numerous golden chloroplasts was present, and the cingulum made 1.5 turns around the cell. The concentration of C. geminatum ranged from 10² to greater than 10⁷ cells l⁻¹ during the bloom period. Nutrient concentration ranges during the bloom were 1.29–81.00 μM NO₃, 0.14–12.14 μM NO₂, 0.21–6.29 μM NH₄, 0.23–6.26 μM PO₄ and 3.29–171.43 μM SiO₃, respectively. Total biomass expressed in terms of chlorophyll a ranged from 2.44 to 135.45 μg l⁻¹, with an average 19.9 μg l⁻¹ in surface water throughout the PRE. Two main clusters corresponding to the water sectors were defined with multivariate analysis (cluster and nMDS). Based on the composition and abundance of phytoplankton, spatial variations were observed at a significant level (ANOSIM, R = 0.44, P < 0.01). Although the pairwise correlation analysis detected no significant effect of any single environmental variable on the abundance of C. geminatum, the multivariate analysis (BIO-ENV) between biotic and abiotic variables resulted in the best variables combination with all measured factors involved (temperature, salinity, turbidity, NO₃, NO₂, NH₄, PO₄ and SiO₃) which showed a combined effect during the bloom of C. geminatum in the Pearl River Estuary (ρ_w = 0.477).     

Light acclimation of photosynthesis in three charophyte species

The main aim of this study was to investigate if the charophyte species Chara baltica, Chara canescens (two populations from the Baltic Sea (BS) and the Gulf of Korinth, Greece (GK)), and Lamprothamnium papulosum exhibit different acclimation capacities to irradiance. Growth, photosynthesis and pigment content were examined in the laboratory under six irradiance conditions (35–500 μmol photons m⁻² s⁻¹). Growth experiments showed increasing growth rates from 35 μmol photons m⁻² s⁻¹ (∼10 mg fresh weight (FW)) up to 70 μmol photons m⁻² s⁻¹ (∼20 mg FW) in C. baltica, from 35 μmol photons m⁻² s⁻¹ (∼15 mg FW) up to 380 μmol photons m⁻² s⁻¹ (∼145 mg FW) in C. canescens (BS), and up to the highest growth irradiance in algae of L. papulosum (35 μmol: ∼5 mg FW; 500 μmol: ∼20 mg FW). The species were tested for their ability to acclimate to different growth irradiances (E_g) by calculating P_max (maximum photosynthesis rate at saturating irradiances), α (the efficiency of light utilization at limiting irradiance), and E_k (the light saturation point of photosynthesis, P_max/α). All species exhibited increasing P_max with increasing E_g. Whereas both populations of C. canescens increased α with increasing E_g, L. papulosum and C. baltica did not acclimate α at all. E_k, the irradiance at which photosynthesis ceased to be light-limited, was constant for all Chara species within the range of irradiances tested. Chl a/Chl b ratios of all species were constant over the whole range of E_g. Chl a/carotenoid ratios were constant in C. baltica, whereas Chl a/carotenoid ratios in L. papulosum and C. canescens (BS) decreased from 250 and 70 μmol photons m⁻² s⁻¹ upwards, respectively. Pigmentation analysis showed that Chl a/carotenoid acclimation was mainly caused by species-specific capacity to raise the content of lutein and carotene (C. canescens (BS), C. canescens (GK)) and xanthophyll cycle pigments (XCP; L. papulosum). The non-photochemical quenching (NPQ) capacities of L. papulosum, C. canescens (BS), and C. canescens (GK) were dependent from preacclimation status of algae, whereas NPQ of C. baltica was independent from growth irradiance.Our results indicate that C. baltica and C. canescens (BS) were light saturated within the chosen irradiances, whereas C. canescens (GK) and L. papulosum did not reach their limits of high-light acclimation. The photosynthetic pigments lutein, α- and β-carotene are suggested to act as photo-protective pigments in L. papulosum and C. canescens.     

Thresholds for morphological response to light reduction for four tropical seagrass species

Seagrasses worldwide are highly vulnerable to, and at increasing risk from reduced light availability, and robust light thresholds are required for evaluating future impacts of changing light conditions. We tested the morphological response (shoot density and growth) of four Indo-West Pacific seagrass species (Cymodocea serrulata, Halodule uninervis, Halophila ovalis and Zostera muelleri) to six daily light levels ranging from 0 to 23 mol m⁻² d⁻¹ (0–70% surface irradiance) in cool (∼23 °C) and warm temperatures (∼28 °C) over 14 weeks. The impact of light limitation on shoot densities and growth rates was higher at warm than at cool temperatures, and for Z. muelleri and H. ovalis than for C. serrulata and H. uninervis, in terms of both the time taken for the low light treatment to take effect and the predicted time to shoot loss (e.g. 17–143 days at 0 mol m⁻² d⁻¹). Using fitted curves we estimated temperature-dependent thresholds (with estimates of uncertainty) for 50% and 80% protection of growth and shoot density, defined here as “potential light thresholds” in recognition that they were derived under experimental conditions. Potential light thresholds that maintained 50% and 80% of seagrass shoot density fell within the ranges 1.1–5.7 mol m⁻² d⁻¹ and 3.8–10.4 mol m⁻² d⁻¹, respectively, depending on temperature and species. Light thresholds calculated in separate in situ studies for two of the same species produced comparable results. We propose that the upper (rounded) values of 6 mol m⁻² d⁻¹ and 10 mol m⁻² d⁻¹ can be used as potential light thresholds for protecting 50% and 80% of shoot density for these four species over 14 weeks. As management guidelines should always be more conservative than thresholds for biological declines, we used error estimates to provide a quantitative method for converting potential light thresholds into guidelines that satisfy this criterion. The present study demonstrates a new approach to deriving potential light thresholds for acute impacts, describes how they can be applied in management guidelines and quantifies the timescales of seagrass decline in response to light limitation. This method can be used to further quantify cumulative impacts on potential light thresholds.     

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R > 0.92 (p < 0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (L_max) and the semi-optimum light intensity range (L_s-opt) that resulted in the maximum growth rate (μ_max) and ≥80% μ _max values of the four species/phylotypes, respectively, were as follows: (1) the L_max and L_s-opt of G. australes were 208 μmol photons m⁻² s⁻¹ and 91–422 μmol photons m⁻² s⁻¹, respectively; (2) those of G. scabrosus were 252 and 120–421 μmol photons m⁻² s⁻¹, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75–430 μmol photons m⁻² s⁻¹, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73–427 μmol photons m⁻² s⁻¹, respectively. All four Gambierdiscus species/phylotypes required approximately 10 μmol photons m⁻² s⁻¹ to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1 m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer’s Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058–0.119 m⁻¹. The values 1700 μmol photons m⁻² s⁻¹, 500 μmol photons m⁻² s⁻¹, and 200 μmol photons m⁻² s⁻¹ were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of L_max and L_s-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12–38 m and 12–54 m, 1–16 m and 1–33 m, and 0 m and 0–16 m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.     

Differential interactions between the nematocyst-bearing mixotrophic dinoflagellate Paragymnodinium shiwhaense and common heterotrophic protists and copepods: Killer or prey

To investigate interactions between the nematocyst-bearing mixotrophic dinoflagellate Paragymnodinium shiwhaense and different heterotrophic protist and copepod species, feeding by common heterotrophic dinoflagellates (Oxyrrhis marina and Gyrodinium dominans), naked ciliates (Strobilidium sp. approximately 35 μm in cell length and Strombidinopsis sp. approximately 100 μm in cell length), and calanoid copepods Acartia spp. (A. hongi and A. omorii) on P. shiwhaense was explored. In addition, the feeding activities of P. shiwhaense on these heterotrophic protists were investigated. Furthermore, the growth and ingestion rates of O. marina, G. dominans, Strobilidium sp., Strombidinopsis sp., and Acartia spp. as a function of P. shiwhaense concentration were measured. O. marina, G. dominans, and Strombidinopsis sp. were able to feed on P. shiwhaense, but Strobilidium sp. was not. However, the growth rates of O. marina, G. dominans, Strobilidium sp., and Strombidinopsis sp. feeding on P. shiwhaense were very low or negative at almost all concentrations of P. shiwhaense. P. shiwhaense frequently fed on O. marina and Strobilidium sp., but did not feed on Strombidinopsis sp. and G. dominans. G. dominans cells swelled and became dead when incubated with filtrate from the experimental bottles (G. dominans + P. shiwhaense) that had been incubated for one day. The ingestion rates of O. marina, G. dominans, and Strobilidium sp. on P. shiwhaense were almost zero at all P. shiwhaense concentrations, while those of Strombidinopsis sp. increased with prey concentration. The maximum ingestion rate of Strombidinopsis sp. on P. shiwhaense was 5.3 ng C predator⁻¹d⁻¹ (41 cells predator⁻¹d⁻¹), which was much lower than ingestion rates reported in the literature for other mixotrophic dinoflagellate prey species. With increasing prey concentrations, the ingestion rates of Acartia spp. on P. shiwhaense increased up to 930 ng C ml⁻¹ (7180 cells ml⁻¹) at the highest prey concentration. The highest ingestion rate of Acartia spp. on P. shiwhaense was 4240 ng C predator⁻¹d⁻¹ (32,610 cells predator⁻¹d⁻¹), which is comparable to ingestion rates from previous studies on other dinoflagellate prey species calculated at similar prey concentrations. Thus, P. shiwhaense might play diverse ecological roles in marine planktonic communities by having an advantage over competing phytoplankton in anti-predation against potential protistan grazers.     

Interactive effects of salinity and water depth on the growth of Melaleuca ericifolia Sm. (Swamp paperbark) seedlings

Melaleuca ericifolia Sm. (Swamp paperbark) is a common tree species in freshwater and brackish wetlands in southern and eastern Australia. The survival of this species in many wetlands is now threatened by increased salinity and inappropriate water regimes. We examined the response of 5-month-old M. ericifolia seedlings to three water depths (exposed, waterlogged and submerged) at three salinities (2, 49 and 60 dS m⁻¹). Increasing water depth at the lowest salinity did not affect survival, but strongly inhibited seedling growth. Total biomass, leaf area and maximum root length were highest in exposed plants, intermediate in waterlogged plants and lowest in submerged plants. Although completely submerged plants survived for 10 weeks at the lowest salinity, they demonstrated negative growth rates and were unable to extend their shoots above the water surface. At the higher salinities, M. ericifolia seedlings were intolerant of waterlogging and submergence: all plants died after 9 weeks at 60 dS m⁻¹. Soil salinities increased over time, and by Week 10, exceeded external water column salinities in both the exposed and waterlogged treatments. In exposed sediment, ∼90% of plants survived for 10 weeks at 60 dS m⁻¹ even though soil salinities reached ∼76 dS m⁻¹. No mortality occurred in the exposed plants at 49 dS m⁻¹, and small but positive relative growth rates were recorded at Week 10. We conclude that at low salinities M. ericifolia seedlings are highly tolerant of sediment waterlogging, but are unlikely to tolerate prolonged submergence. However, at the higher salinities, M. ericifolia seedlings are intolerant of waterlogging and submergence and died rapidly after 5 weeks exposure to this combination of environmental stressors. This research demonstrates that salinity may restrict the range of water regimes tolerated by aquatic plants.     

Germination fluctuation of toxic Alexandrium fundyense and A. pacificum cysts and the relationship with bloom occurrences in Kesennuma Bay,Japan

While cyst germination may be an important factor for the initiation of harmful/toxic blooms, assessments of the fluctuation in phytoplankton cyst germination, from bottom sediments to water columns, are rare in situ due to lack of technology that can detect germinated cells in natural bottom sediments. This study introduces a simple mesocosm method, modeled after previous in situ methods, to measure the germination of plankton resting stage cells. Using this method, seasonal changes in germination fluxes of toxic dinoflagellates resting cysts, specifically Alexandrium fundyense (A. tamarense species complex Group I) and A. pacificum (A. tamarense species complex Group IV), were investigated at a fixed station in Kesennuma Bay, northeast Japan, from April 2014 to April 2015. This investigation was conducted in addition to the typical samplings of seawater and bottom sediments to detect the dinoflagellates vegetative cells and resting cysts. Bloom occurrences of A. fundyense were observed June 2014 and February 2015 with maximum cell densities reaching 3.6 × 10⁶ cells m⁻² and 1.4 × 10⁷ cells m⁻², respectively. The maximum germination fluxes of A. fundyense cysts occurred in April 2014 and December 2014 and were 9.3 × 10³ cells m⁻² day⁻¹ and 1.4 × 10⁴ cells m⁻² day⁻¹, respectively. For A. pacificum, the highest cell density was 7.3 × 10⁷ cells m⁻² during the month of August, and the maximum germination fluxes occurred in July and August, reaching 5.8 × 10² cells m⁻² day⁻¹. Thus, this study revealed the seasonal dynamics of A. fundyense and A. pacificum cyst germination and their bloom occurrences in the water column. Blooms occurred one to two months after peak germination, which strongly suggests that both the formation of the initial population by cyst germination and its continuous growth in the water column most likely contributed to toxic bloom occurrences of A. fundyense and A. pacificum in the bay.     

Comparison of salinity tolerance of three Atriplex spp. in well-watered and drying soils

Members of the Chenopodiaceae are well adapted to both salt and drought stress and can serve as model species to understand the mechanisms of tolerance in plants. We grew Atriplex hortensis (ATHO), A. canescens (ATCA), and A. lentiformis (ATLE) along a NaCL salinity gradient under non-water-limited conditions and in drying soils in greenhouse experiments. The species differed in photosynthetic carbon fixation pathway, capacity for sodium uptake, and habitat preferences. Under non-water-limited conditions, ATLE (C4) maintained high growth rates up to 30 g L⁻¹ NaCl. ATHO (C3) had lower growth than ATLE at high salinities, while ATCA (C4) grew more slowly than either ATLE or ATHO and showed no net growth above 20 g L⁻¹ NaCl. ATHO and ATLE accumulated twice as much sodium in their shoots as ATCA, but all three species had increasing sodium levels at higher salinities. Potassium, magnesium and calcium levels were relatively constant over the salinity gradient. All three species showed marked accumulation of chloride across the salinity gradient, whereas nitrate, phosphorous and sulfate decreased with salinity. The effect of drought was simulated by growing plants in sealed pots with an initial charge of water plus NaCl, and allowing them to grow to the end point at which they no longer were able to extract water from the soil solution. Drought and salinity were not additive stress factors for Atriplex spp. in this experiment. NaCl increased their ability to extract water from the soil solution compared to fresh water controls. ATLE showed increased shoot dry matter production and increased water use efficiency (WUE) as initial salinity levels increased from 0 to 30 g L⁻¹ NaCl, whereas dry matter production and WUE peaked at 5 g L⁻¹ for ATHO and ATCA. Final soil moisture salinities tolerated by species were 85 g L⁻¹, 55 g L⁻¹ and 160 g L⁻¹ NaCl for ATHO, ATCA and ATLE, respectively. C4 photosynthesis and sodium accumulation in shoots were associated with high drought and salt tolerance.     

Purification and characterization of yellow laccase from Trametes hirsuta MTCC-1171 and its application in synthesis of aromatic aldehydes

A yellow laccase from the culture filtrate of Trametes hirsuta MTCC-1171 has been purified. The purification methods involved concentration of the culture filtrate by ammonium sulphate precipitation and an anion exchange chromatography on diethylaminoethyl cellulose. The sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and native polyacrylamide gel electrophoresis gave single protein band indicating that the enzyme preparation was pure. The molecular mass of the enzyme determined from SDS-PAGE analysis was 55.0 kDa. Using 2,6-dimethoxyphenol, 2,2′[azino-bis-(3-ethylbonzthiazoline-6-sulphonic acid) diammonium salt] and 3,5-dimethoxy-4-hydroxybenzaldehyde azine as the substrates, the K_m, k_cat and k_cat/K_m values of the laccase were found to be 420 μM, 13.04 s⁻¹, 3.11 × 10⁴ M⁻¹ s⁻¹, 225 μM, 13.03 s⁻¹, 1.3 × 10⁵ M⁻¹ s⁻¹ and 100 μM, 13.04 s⁻¹, 5.8 × 10⁴ M⁻¹ s⁻¹, respectively. The pH and temperature optima were 4.5 and 60 °C, respectively while pH and temperature stabilities were pH 4.5 and 50 °C. The activation energy for thermal denaturation of the enzyme was 18.6 kJ/mol/K. The purified laccase has yellow colour and does not show absorption band around 610 nm like blue laccases. The purified laccase transforms toluene, 3-nitrotoluene, 4-nitrotoluene, 3-chlorotoluene, 4-chlorotoluene and 3,4-dimethoxytoluene to benzaldehyde, 3-nitrobenzaldehyde, 4-nitrobenzaldehyde, 3-chlorobenzaldehyde, 4-chlorobenzaldehyde and 3,4-dimethoxybenzaldehyde in the absence of mediator molecules in high yields.     

Growth and bioactive secondary metabolites of arctic Protoceratium reticulatum (Dinophyceae)

Harmful algal blooms are mainly caused by marine dinoflagellates and are known to produce potent toxins that may affect the ecosystem, human activities and health. Such events have increased in frequency and intensity worldwide in the past decades. Numerous processes involved in Global Change are amplified in the Arctic, but little is known about species specific responses of arctic dinoflagellates. The aim of this work was to perform an exhaustive morphological, phylogenetical and toxinological characterization of Greenland Protoceratium reticulatum and, in addition, to test the effect of temperature on growth and production of bioactive secondary metabolites. Seven clonal isolates, the first isolates of P. reticulatum available from arctic waters, were phylogenetically characterized by analysis of the LSU rDNA. Six isolates were further characterized morphologically and were shown to produce both yessotoxins (YTX) and lytic compounds, representing the first report of allelochemical activity in P. reticulatum. As shown for one of the isolates, growth was strongly affected by temperature with a maximum growth rate at 15 °C, a significant but slow growth at 1 °C, and cell death at 25 °C, suggesting an adaptation of P. reticulatum to temperate waters. Temperature had no major effect on total YTX cell quota or lytic activity but both were affected by the growth phase with a significant increase at stationary phase. A comparison of six isolates at a fixed temperature of 10 °C showed high intraspecific variability for all three physiological parameters tested. Growth rate varied from 0.06 to 0.19 d⁻¹, and total YTX concentration ranged from 0.3 to 15.0 pg  YTX cell⁻¹ and from 0.5 to 31.0 pg YTX cell⁻¹ at exponential and stationary phase, respectively. All six isolates performed lytic activity; however, for two isolates lytic activity was only detectable at higher cell densities in stationary phase.     

Seasonal variations in photosynthetic irradiance response curves of macrophytes from a Mediterranean coastal lagoon

The main photosynthesis and respiration parameters (dark respiration rate, light saturated production rate, saturation irradiance, photosynthetic efficiency) were measured on a total of 23 macrophytes of the Thau lagoon (2 Phanerogams, 5 Chlorophyceae, 10 Rhodophyceae and 6 Phaeophyceae). Those measurements were performed in vitro under controlled conditions, close to the natural ones, and at several seasons. Concomitantly, measurements of pigment concentrations, carbon, phosphorous and nitrogen contents in tissues were performed. Seasonal intra-specific variability of photosynthetic parameters was found very high, enlightening an important acclimatation capacity. The highest photosynthetic capacities were found for Chlorophyceae (e.g. Monostroma obscurum thalli at 17 °C, 982 μmol O₂ g⁻¹ dw h⁻¹ and 9.1 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹, respectively for light saturated net production rate and photosynthetic efficiency) and Phanerogams (e.g. Nanozostera noltii leaves at 25 °C, 583 μmol O₂ g⁻¹ dw h⁻¹ and 2.6 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹ respectively for light saturated net production rate and photosynthetic efficiency). As expected, species with a high surface/volume ratio were found to be more productive than coarsely branched thalli and thick blades shaped species. Contrary to R_d (ranging 6.7–794 μmol O₂ g⁻¹ dw h⁻¹, respectively for Rytiphlaea tinctoria at 7 °C and for Dasya sessilis at 25 °C) for which a positive relationship with water temperature was found whatever the species studied, the evolution of P/I curves with temperature exhibited different responses amongst the species. The results allowed to show summer nitrogen limitation for some species (Gracilaria bursa-pastoris and Ulva spp.) and to propose temperature preferences based on the photosynthetic parameters for some others (N. noltii, Zostera marina, Chaetomorpha linum).