Allelopathic interactions between Prorocentrum micans and Skeletonema costatum or Karenia mikimotoi in laboratory cultures

Algal allelopathy is an ecological/physiological phenomenon that has focused attention on the interactions among algae and the production of algal toxins. We investigated the allelopathic interactions between the dinoflagellate genus Prorocentrum micans and diatom genus Skeletonema costatum and between P. micans and dinoflagellate genus Karenia mikimotoi using bi-algal cultures. Because the effects were species-specific and size-dependent, we evaluated the effect of different initial densities. At low densities of P. mieans and high densities of S. costatum inoculated into the same medium, the growth of R rnieans was weakly restrained, whereas the growth of S. costatum was significantly suppressed. S. costatum and K. mikimotoi were strongly inhibited by P. micans, in both the bi-algal cultures and enriched filtrates. Direct cell-to-cell contact was not necessary to gain a competitive advantage, thus, our results suggest that P. micans inhibited the growth of S. costatum and K. mikimotoi by the release of allelochemical（s）. Last, a mathematical model was used to simulate growth and interactions between P. micans and S. eostatum and between P. micans and K. mikimotoi in bi-algal cultures.

Effects of NO and Different Media on the Growth of Prorocentrum micans

Prorocentrum micans was cultivated in different media including f/2, f/4, f/8, f/20, f/25, f/50, f/100 and f/200. Results showed that media influence the growth of P. micans. The biomass of P. micans in rich nutrition medium was much higher than in poor nutrition medium. Nitric oxide can promote or inhibit the growth of P. micans in all media. Nitric oxide at the concentration of 1.4 × 10^- 6 mol L^-1 promoted the growth of P. micans significantly when added only once during the cultivation. When added twice a day, nitric oxide at the concentration of 1.4 × 10^- 9 mol L^-1 promoted the growth of P. micans significantly, while nitric oxide at the concentrations of 1.4 × 10^- 5 mol L ^-1 and 1.4 × 10^- 6 mol L ^-1 inhibited the growth. Therefore, nitric oxide, media and the ways to add nitric oxide influenced the growth of P. micans respectively.

Size-fractionated phytoplankton biomass in autumn of the Changjiang(Yangtze) River Estuary and its adjacent waters after the Three Gorges Dam construction

A cruise was undertaken from 3rd to 8th November 2004 in Changjiang(Yangtze) River Estuary and its adjacent waters to investigate the spatial biomass distribution and size composition of phytoplankton.Chlorophyll-a(Chl-a) concentration ranged 0.42-1.17 μg L-1 and 0.41-10.43 μg L-1 inside and outside the river mouth,with the mean value 0.73 μg L-1 and 1.86 μg L-1,respectively.Compared with the Chl-a concentration in summer of 2004,the mean value was much lower inside,and a little higher outside the river mouth.The maximal Chl-a was 10.43 μg L-1 at station 18(122.67°E,31.25°N),and the region of high Chl-a concentration was observed in the central survey area between 122.5°E and 123.0°E.In the stations located east of 122.5°E,Chl-a concentration was generally high in the upper layers above 5 m due to water stratification.In the survey area,the average Chl-a in sizes of >20 μm and <20 μm was 0.28 μg L-1 and 1.40 μg L-1,respectively.High Chl-a concentration of <20 μm size-fraction indicated that the nanophytoplankton and picophytoplankton contributed the most to the biomass of phytoplankton.Skeletonema costatum,Prorocentrum micans and Scrippsiella trochoidea were the dominant species in surface water.The spatial distribution of cell abundance of phytoplankton was patchy and did not agree well with that of Chl-a,as the cell abundance could not distinguish the differences in shape and size of phytoplankton cells.Nitrate and silicate behaved conservatively,but the former could probably be the limitation factor to algal biomass at offshore stations.The distribution of phosphate scattered considerably,and its relation to the phytoplankton biomass was complicated.

Identification of the toxic compounds produced by Sargassum thunbergii to red tide microalgae

The inhibitory effects of methanol extracts from the tissues of three macroalgal species on the growths of three marine red tide microalgae were assessed under laboratory conditions.Extracts of Sargassum thunbergii(Mertens ex Roth) Kuntz tissue had stronger inhibitory effects than those of either Sargassum pallidum(Turner) C.Agardh or Sargassum kjellmanianum Yendo on the growths of Heterosigma akashiwo(Hada) Hada,Skeletonema costatum(Grev.) Grev,and Prorocentrum micans Ehrenberg.Methanol extracts of S.thunbergii were further divided into petroleum ether,ethyl acetate,butanol,and distilled water phases by liquid-liquid fractionation.The petroleum ether and ethyl acetate fractions had strong algicidal effects on the microalgae.Gas chromatography-mass spectrometry analyses of these two phases identified nine fatty acids,most of which were unsaturated fatty acids.In addition,pure compounds of four of the nine unsaturated fatty acids had effective concentrations below 5 mg/L.Therefore,unsaturated fatty acids are a component of the allelochemicals in S.thunbergii tissue.

Effects of Nitrogen Sources and Concentrations on the Growth of Different Phytoplankton Taxa

Effects of different nitrogen(N)compounds and concentrations on the growth of the three different phytoplankton taxa,Skeletonema costatum(Bacillariophyceae),Prorocentrum micans(Dinophyceae),and Chattonella marina(Raphidophyceae),were investigated.The Monod equation was applied to examine effects of N concentrations on the growth of algal cells.Results showed that nitrate(NO_(3)-N)and urea served as good N sources for the three phytoplankton taxa.S.costatum grew well on all of the seven N sources.C.marina can effectively use the two amino acids,glycine(Gly)and serine(Ser),however cannot utilize alanine(Ala),threonine(Thr),and asparaginic acid(Asp).P.micans cannot grow in five amino acid substrates.All of the three phytoplankton taxa grew well under different proportions of urea-N,and C.marina grew significantly better in medium with both NO_(3)-and urea-N.The values of maximum growth rate(μmax)and half-saturation nutrient concentration(KS)for NO_(3)-N were 0.71 divisions d−1 and 53.55μmolL^(−1) for S.costatum,0.67 divisions d−1 and 23.31μmolL^(−1) for P.micans,and 0.23 divisionsd−1 and 17.57μmolL^(−1) for C.marina,respectively.The results suggested that S.costatum had a high N demand for growth,and was capable of using wide ranges of N compounds.The strategy of N utilization for S.costatum may make this species an advantage in N-enriched sea areas especially the dissolved organic nitrogen(DON)rich coastal waters,which might be the reason why S.costatum widely distributes in the cosmo-politan coastal and estuarine sea areas.

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide(DMS),acrylic acid(AA),and dimethylsulfoniopropionate(DMSP) during growth of three microalgae:Prorocentrum micans,Gephyrocapsa oceanica,and Platymonas subcordiformis.The DMSP,AA,and DMS concentrations in culture media varied significantly among algal growth stages,with the highest concentrations in the late stationary growth stage or the senescent stage.In the stationary growth stage,the average DMSP concentration per cell in P.micans(0.066 5 pmol/cell) was 1.3 times that in G.oceanica(0.049 5 pmol/cell) and 20.2 times that in P.subcordiformis(0.003 29 pmol/cell).The average concentrations of AA were0.044 6,0.026 9,and 0.003 05 pmol/cell in P.micans,G.oceanica,and P.subcordiformis,respectively,higher than the concentrations of DMS(0.272,0.497,and 0.086 2 fmol/cell,respectively).There were significant positive correlations between cell density and AA,DMSP,and DMS concentrations.The ratios of DMS/AA and AA/(DMSP+AA) in the three algae differed significantly over the growth cycle.In all three microalgae,the DMS/AA ratios were less than 25%during the growth period,suggesting that the enzymatic cleavage pathway,which generates DMS,was not the main DMSP degradation pathway.The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence.In all three microalgae,the AA/(DMSP+AA) ratio(degradation ratio of DMSP) decreased during the exponential growth phase,and then increased.The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.

Growth inhibition to three red tide microalgae by extracts of Ulva pertusa

Growth inhibition effect of different concentration of distilled water extract and four polar organic solvent (methanol, acetone, ether and chloroform) extracts of Ulva pertusa on three typical red tide microalgae (Heterosigma akashiwo, Alexandrium tamarense and Prorocentrum micans) were inves- tigated. Liquid-liquid fractionation and HPLC analysis for methanol extract of U. pertusa were carried out. Growth of the three microalgae was significantly inhibited by the distilled water extract of U. pertusa at relatively higher concentration. However, the cells of the three microalgae did not die completely even at high concentration. Methanol extract of U. pertusa showed the highest growth inhibition on the three mi- croalgae, and all the cells of the three microalgae were killed at relatively high concentration. The other three organic solvent extracts of U. pertusa had no apparent effect on the three microalgae. The results of bioassays and HPLC analysis suggested that the inhibitory substances in U. pertusa to the microalgal growth had relatively high polarities. H. akashiwo was the most sensitive one while A. tamarense was the most tolerant one to the growth inhibitory substances.

Screening and isolation of the algicidal compounds from marine green alga Ulva intestinalis

Twenty species of seaweed were collected from the coast of Zhejiang, China, extracted with ethanol, and screened for algicidal activity against red tide microalgae H eterosigma akashiwo and Prorocentrum micans. Inhibitory eff ects of fresh and dried tissues of green alga U lva intestinalis were assessed and the main algicidal compounds were isolated, purifi ed, and identifi ed. Five seaweed species, U. intestinalis, U. fasciata, Grateloupia romosissima, Chondria crassicaulis, and Gracilariopsis lemaneiformis, were investigated for their algicidal activities. Fresh tissues of 8.0 and 16.0 mg/m L of U. intestinalis dissolved in media signifi cantly inhibited growth of H. akashiwo and P. micans, respectively. Dried tissue and ethyl acetate(Et OAc) extracts of U. intestinalis at greater than 1.2 and 0.04 mg/m L, respectively, were fatal to H. akashiwo, while its water and Et OAc extracts in excess of 0.96 and 0.32 mg/m L, respectively, were lethal to P. micans. Three algicidal compounds in the Et OAc extracts were identifi ed as 15-ethoxy-(6z,9z,12z)-hexadecatrienoic acid(I),(6E,9E,12E)-(2-acetoxy- β- D-glucose)-octadecatrienoic acid ester(II) and hexadecanoic acid(III). Of these, compound II displayed the most potent algicidal activity with IC_(50) values of 4.9 and 14.1 μg/m L for H. akashiwo and P. micans, respectively. Compound I showed moderate algicidal activity with IC_(50) values of 13.4 and 24.7 μg/m L for H. akashiwo and P. micans, respectively. These fi ndings suggested that certain macroalgae or products therefrom could be used as ef fective biological control agents against red tide algae.

Interspecific Competition Between Levanderina fissa(Dinophyceae) and the Other Three Microalgal Species in Co-Cultures

Growth interactions between the dinoflagellate Levanderina fissa (=Gyrodinium instriatum) and the other three algal bloom species Skeletonema costatum (Bacillariophyceae), Prorocentrum micans (Dinophyceae), and Chattonella marina (Raphido- phyceae) were studied using bi- or tri-algal co-cultures under laboratory conditions. The Lotka-Volterra model was applied to esti- mate the interspecific competition between them. Results showed that the growth of L. fissa was significantly suppressed in both bi-algal and tri-algal cultures, and the maximum cell densities in co-cultures were only between 21% and 69% of that in mono-cul- ture. L. fissa outcompeted P. micans;however, its effects on S. costatum and C. marina were generally less than the effects of their intraspecific competitions based on the competition coefficients. Contrarily, the inhibitory effects of S. costatum and C. marina on L. fissa were much greater. S. costatum outcompeted the other three species in all co-cultures, and its competition coefficients on the other species were more than 10000 times higher than the opposite. The growth of C. marina was suppressed by S. costatum;how- ever, it outcompeted L. fissa and P. micans. The growth of P. micans was most strongly inhibited in co-cultures, and the maximum cell densities in co-cultures were less than 15% of that in mono-species culture. The results suggested that L. fissa had few advan- tages in the interspecific competition. The adaptation to low salinity and the utilization of organic phosphorus might be the important reasons for L. fissa to take the competitive predominance in brackish estuarine environments.