Understan ding the mechanism of harmful algal bloom formation is vital for effectively preventing algal bloom outbreaks in coastal environments.Karenia spp.blooms in the East China Sea show a significant correlation w...Understan ding the mechanism of harmful algal bloom formation is vital for effectively preventing algal bloom outbreaks in coastal environments.Karenia spp.blooms in the East China Sea show a significant correlation with nutrient regimes.However,the impact of key components of nutrients,especially dissolved organic nitrogen(DON),on the blooms of Karenia spp.is not clear.Quantitative research is still lacking.In this study,the cruise observations,field mesocosm-flask culture experiments,and a multinitrogen-tri-phytoplankton-detritus model(NTPD) are combined to reveal the quantitative influence of nutrient regimes on the shift of Prorocentrum donghaiense and Karenia spp.in the East China Sea.It has a synchronism rhythm of diatom-P.donghaienseKarenia spp.-diatom loop in the field culture experiment,which is consistent with the results of the cruise observation.The results showed that the processes of terrigenous DON(TeDON) and dissolved inorganic nitrogen(DIN:NO_(3)^(-)-N,NH_(4)^(+)-N) absorption promoted P.donghaiense to become the dominant algae in the community;whereas the processes of DON from P.donghaiense absorption promoted Karenia spp.to become the dominant algae in ambient DIN exhaustion.In addition,the three-dimensional fluorescence components of humus C,tyrosine and fulvic acid can indicate the processes of growth and extinction of P.donghaiense and Karenia spp.,respectively.This study infers that P.donghaiense and Karenia spp.regime shift mechanism associated with the nutrient regime in coastal waters,which provides a scientific basis for the environmental management of coastal eco system health.展开更多
Prorocentrum donghaiense is one of the most common red tide causative dinoflagellates in the Changjiang （Yangtze） River Estuary and the adjacent area of the East China Sea. It causes large-scale blooms in late sprin...Prorocentrum donghaiense is one of the most common red tide causative dinoflagellates in the Changjiang （Yangtze） River Estuary and the adjacent area of the East China Sea. It causes large-scale blooms in late spring and early summer that lead to widespread ecologic and economic damage. A means for distinguish- ing dinoflagellate blooms from diatom （Skeletonema costatum） blooms is desired. On the basis of measure- ments of remote sensing refectance [Rrs（λ）] and inherent optical parameters, the potential of using a mul- tispectral approach is assessed for discriminating the algal blooms due to P. donghaiense from those due to S. costatum. The behavior of two reflectance ratios [R1 = Rrs（560）/Rrs（532） and Re = Rrs（708）/Rrs（665）], suggests that differentiation of P. donghaiense blooms from diatom bloom types is possible from the current band setup of ocean color sensors. It is found that there are two reflectance ratio regimes that indicate a bloom is dominated by P. donghaiense; （1） R1 〉 1.55 and R2 〈 1.0 or （2） R1 〉 1.75 and R2 ≥ 1.0. Various sensitivity analyses are conducted to investigate the effects of the variation in varying levels of chlorophyll concentration and colored dissolved organic matter （CDOM） as well as changes in the backscattering ratio （bbp/bp） on the efficacy of this muitispectral approach. Results indicate that the intensity and inherent op- tical properties of the algal species explain much of the behavior of the two ratios. Although backscattering influences the amplitude of Rrs（λ）, especially in the 530 and 560 nm bands, the discrimination between P. donghaiense and diatoms is not significantly affected by the variation of bbp/bp. Since aCDOM （440） in coastal areas of the ECS is typically lower than 1.0 m-1 in most situations, the presence of CDOM does not interfere with this discrimination, even as SCDOM varies from 0.01 to 0.026 nm-1. Despite all of these effects, the dis- crimination of P. donghaiense blooms from diatom blooms based on multispectral measurements of Rrs（λ） is feasible.展开更多
Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrurn donghaiense at the exponential growth, stationary and decline stages into 〈0.45 μm filtrate, 100 kDa-0.45 μm...Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrurn donghaiense at the exponential growth, stationary and decline stages into 〈0.45 μm filtrate, 100 kDa-0.45 μm, 1%100 kDa and 1-10 kDa retentate and 〈1 kDa ultrafiltrate fractions. The fluorescence properties of different molecular weights of dissolved organic matter （DOM） were measured by excitation-emission matrix spectra. Protein-like and humic-like fluorophores were observed in the DOM produced by P. donghaiense. The central positions of protein-like fluorophores showed a red shift with prolonged growth duration, shifting from tyrosine-like properties at the exponential growth stage to tryptophan-like properties at the stationary and decline stages. The excitation wavelengths of protein-like fluorophores exhibited some change in the exponential growth and stationary stages with increased molecular size, but showed little change in the decline stage. However, the emission wavelengths in the decline stage exhibited a blue shift. Very distinct C type and A type peaks in humic-like fluorophores were observed. With a prolonged culture time, the intensities of both of the peaks became strong and the excitation wavelengths of peak A showed a red shift, while the A：C ratios fell. More than 94% of fluorescent DOM was in the lower than 1 kDa molecular weight fraction.展开更多
Most of reported harmful algal blooms(HABs)of microalgae(75%)have been caused by dinoflagellates.Studies on the negative effects of HABs have generally focused on animals,valuable organisms in particular,and environme...Most of reported harmful algal blooms(HABs)of microalgae(75%)have been caused by dinoflagellates.Studies on the negative effects of HABs have generally focused on animals,valuable organisms in particular,and environmental factors such as dissolved oxygen and nutrients,but relatively fewer on community level,particularly that using metagenomic approach.In this study,we reported an investigation on the effects of a HAB caused by the dinoflagellate Prorocentrum donghaiense on the species diversity and community structure of the dinoflagellate sub-community via a pyrosequencing approach for the samples taken before,during,and after the bloom season of P.donghaiense in the East China Sea.We sequenced partial 28S rRNA gene of dinoflagellates for the field samples and evaluated the species richness and diversity indices of the dinoflagellate community,as a sub-community of the total phytoplankton.We obtained 800185 valid sequences(categorized into 560 operational taxonomic units,OTUs)of dinoflagellates from 50 samples and found that the biodiversity of dinoflagellate community was significantly reduced during the blooming period in comparison to that in pre-and after-blooming periods,as reflected in the four diversity indices:the species richness expressed as the number of OTUs,Chao1 index,Shannon index(evenness),and Gini-Simpson index.These four indices were all found to be negatively correlated to the cell density of the bloom species P.donghaiense.Correlation analyses also revealed that the P.donghaiense cell abundance was correlated negatively with NO3--N,and NO2--N,but positively with total nitrogen(TN)and total phosphorus(TP).Principal coordinates analysis(PCoA)showed that the community structure of dinoflagellates was markedly different among the different sampling periods,while the redundancy analysis(RDA)revealed P.donghaiense abundance,salinity,NO3--N,and SiO32-were the most four significant factors shaping the dinoflagellate community structure.Our results together demonstrated that HABs caused by the dinoflagellate P.donghaiense could strongly impact the aquatic ecosystem on the sub-community level which the blooming species belongs to.展开更多
Mitochondrial cytochrome b （Cytb）, one of the few proteins encoded by the mitochondrial DNA, plays an important role in transferring electrons. As a mitochondrial gene, it has been widely used for phylogenetic analy...Mitochondrial cytochrome b （Cytb）, one of the few proteins encoded by the mitochondrial DNA, plays an important role in transferring electrons. As a mitochondrial gene, it has been widely used for phylogenetic analysis. Previously, a 949-bp fragment of the coding gene and mRNA editing were characterized from Prorocentrum donghaiense, which might prove useful for resolving P. donghaiense from closely related species. However, the full-length coding region has not been characterized. Ih this study, we used rapid amplification of cDNA ends （RACE） to obtain full-length, 1 124 bp cDNA. Cytb transcript contained a standard initiation codon ATG, but did not have a recognizable stop codon. Homology comparison showed that the P. donghaiense Cytb had a high sequence identity to Cytb sequences from other dinoflagellate species. Phylogenetic analysis placed Cytb from P. donghaiense in the clade of dinoflagellates and it clustered together strongly with that from P. minimum. Based on the full-length sequence, we inferred 32 editing events at different positions, accounting for 2.93% of the Cytb gene. 34.4% （11） of the changes were A to G, 25% （8） were T to C, and 25% （8） were C to U, with smaller proportions of G to C and G to A edits （9.4% （3） and 6.2% （2）, respectively）. The expression level of the Cytb transcript was quantified by real-time PCR with a TaqMan probe at different times during the whole growth phase. The average Cytb transcript was present at 39.277.46 copies of cDNA per cell during the whole growth cycle, and the expression of Cytb was relatively stable over the different phases. These results deepen our understanding of the structure and characteristics of Cytb in P. donghaiense, and confirmed that Cytb in P. donghaiense is a candidate reference gene for studying the expression of other genes.展开更多
Algal blooms caused by Prorocentrum donghaiense occurred frequently in the East China Sea （ECS） during spring in recent years. In this study, a coupled biophysical model was used to hindcast a massive P. donghaiense...Algal blooms caused by Prorocentrum donghaiense occurred frequently in the East China Sea （ECS） during spring in recent years. In this study, a coupled biophysical model was used to hindcast a massive P. donghaiense bloom that occurred in 2005 and to determine the factors influencing bloom initiation and development. The model comprised the Regional Ocean Modeling System tailored for the ECS that utilized a multi-nested configuration and a population dynamics model for 19. donghaiense. Comparisons between simulations and observations revealed that the biological model is capable of reproducing the characteristics of 19. donghaiense growth under different irradiances and phosphorus limitation scenarios. The variation of intracellular phosphorus and the effects of 19. donghaiense on ambient nutrients conditions were also reproduced. The biophysical model hindcasted the hydrodynamics and spatiotemporal distributions of the P. donghaiense bloom reasonably well. Bloom development was consistent with observations reported in earlier studies. The results demonstrate the capability of the model in capturing subsurface incubation during bloom initiation. Then model＇s hindcast solutions were further used to diagnose the factors controlling the vertical distribution. Phosphate appeared to be one of the factors controlling the subsurface incubation, whereas surface wind fields played an important role in determining P. donghaiense distribution. The results highlight the importance of nutrient-limitation as a mechanism in the formation of P. donghaiense subsurface layers and the dispersing of P. donghaiense blooms. This coupled biophysical model should be improved and used to investigate 19. donghaiense blooms occurring in different scenarios.展开更多
Prorocentrum donghaiense is an important harmful algae bloom （HAB） causing creature in China＇s seas, and the conventional visual detection can not cope with long-term monitoring and highthroughput sampling projects...Prorocentrum donghaiense is an important harmful algae bloom （HAB） causing creature in China＇s seas, and the conventional visual detection can not cope with long-term monitoring and highthroughput sampling projects. An assay for P. donghaiense with sandwich hybridization integrated with nuclease protection assay （NPA-SH） was established. Tests with mixed samples and spiked field ones confirmed its good specificity and sensitivity. The cell number of P. donghaiense correlated well with the optical density, and the regression equation is y=4× 10^- 6x＋ 0.694 9, in which x is the cell number, and y is the optical density, with r2=0.953 5. These results show that the NPA-SH method has good feasibility in the detection of P. donghaiense. Results of NPA-SH and microscopy are excellent for each sample. The NPA-SH method was a simple way in quantitative detection of P. donghaiense, and the whole process could be finished in about six hours, which provided a new approach in high-throughput sampling and long-term monitoring of P. donghaiense.展开更多
Harmful algal bloom(HAB)is an ecological disaster to local mariculture.At present,its impact on macrophytes has not been well studied.In this study,we cultivated sexually propagated embryos of S argassum fusiformis-an...Harmful algal bloom(HAB)is an ecological disaster to local mariculture.At present,its impact on macrophytes has not been well studied.In this study,we cultivated sexually propagated embryos of S argassum fusiformis-an edible seaweed-in Prorocentrum donghaiense suspensions at different cell densities(0,0.50×10^(5),0.75×10^(5),1.00×10^(5),and 1.50×10^(5) cells/mL)for 10 days,during which growth and photosynthetic activities of the embryos were determined,and a monocultivation was set up for comparison.Results show that the relative growth rate and photosynthetic activities of the embryos co-cultivated with P.donghaiense were inhibited mostly and significantly in the cell densities of 0.75×10^(5),1.00×10^(5),and 1.50×10^(5) cells/mL,and the inhibitory effects increased in overall with increased cell densities.The maximum relative electron transport rates(rETR max)and apparent photosynthetic efficiency(a)of co-cultivated embryos were all significantly lower than monocultivation ones on the 10 th day.Furthermore,the photosynthetic activity detected by chlorophyll-a fluorescence transient(i.e.,OJIP),the electron transport among electron transfer accepters of PSII(photosystem II)and that from PSII to PSI(photosystem I)was restricted,which is probably responsible for the decreases of rETR max andain the co-cultivated embryos.In addition,parts of the photosynthetic reaction centers of PSII in the co-cultivated embryos were inactivated.Therefore,P.donghaiense bloom could restrain the development and photosynthetic activities of S.fusiformis embryos,reduce the seedlings stock,and eventually hinder the development of S.fusiformis production industry.展开更多
Interactions between Skeletonema costatum (S. costatum) and Prorocentrum donghaiense (P. donghaiiense) were inves-tigated using bi-algal cultures at different concentrations of phosphate (PO4-P) and nitrate/phosphate ...Interactions between Skeletonema costatum (S. costatum) and Prorocentrum donghaiense (P. donghaiiense) were inves-tigated using bi-algal cultures at different concentrations of phosphate (PO4-P) and nitrate/phosphate (N/P) ratios. Experiments were conducted under P-limited conditions and the Lotka-Volterra mathematical model was used to simulate the growth of S. costatum and P. donghaiense in the bi-algal cultures. Both of these two species were inhibited significantly in bi-algal culture. The results of the simulation showed that the inhibitory degree of S. costaum by P. donghaiense was high when the concentration of PO4-P was low (0.1μmolL-1/2 d), but that of P. donghaiense by S. costaum was high with increased PO4-P supply (0.6μmolL-1/2 d). At low concen-tration of PO4-P (0.1μmolL-1/2 d), or high concentration of PO4-P (0.6μmolL-1/2 d) with high N/P ratio (160), the interactions be-tween S. costatum and P. donghaiense were dependent on the initial cell densities of both species. At high concentration of PO4-P (0.6μmolL-1/2 d) with low N/P ratio (25 or 80), S. costatum exhibited a survival strategy superior to that of P. donghaiense. The de-gree of inhibition of P. donghaiense by S. costaum increased with elevated N/P ratio when the medium was supplemented with con-centration 0.1μmolL-1/2 d of PO4-P. The degree of inhibition to P. donghaiense by S. costaum increased with elevated N/P ratio at low concentration of PO4-P (0.1 μmolL-1/2 d). This trend was conversed at high concentration of PO4-P (0.6μmolL-1/2 d). However, the degree of inhibition of S. costaum by P. donghaiense increased with the increased N/P ratio at different PO4-P concentrations (0.1μmolL-1/2 d and 0.6μmolL-1/2 d). These results suggested that both phosphate concentration and N/P ratio affected the competition between S. costaum and P. donghaiense: P. donghaiense is more competitive in environments with low phosphate or high N/P ratio and the influence of N/P ratio on the competition was more significant with lower phosphate concentration.展开更多
Dinofl agellates are the major causative agents of harmful algal blooms in the global ocean and they usually form blooms under conditions of very low dissolved inorganic phosphorus(DIP).However,the mechanisms underpin...Dinofl agellates are the major causative agents of harmful algal blooms in the global ocean and they usually form blooms under conditions of very low dissolved inorganic phosphorus(DIP).However,the mechanisms underpinning the dinofl agellate blooms remain unclear.Here,we quantitatively compared protein expression profi les of a marine dinofl agellate,Prorocentrum donghaiense,grown in inorganic P-replete,P-defi cient,and DIP-and dissolved organic phosphorus(DOP)-resupplied conditions by employing a Tandem Mass Tag(TMT)-based quantitative proteomic approach.Proteins involved in intracellular P reallocation,organic P,and non-P lipid utilization were up-regulated under the P-defi cient condition,while inorganic phosphate transporters varied insignifi cantly.In response to the P resupplementation,nitrogen metabolism,ribosome,porphyrin,and chlorophyll metabolism were up-regulated,while lysosome,and starch and sucrose metabolism were down-regulated.Notably,photosynthesis was up-regulated and secondary metabolism was down-regulated only in the DIP-resupplied cells,whereas amino acid metabolism and vitamin B6 metabolism were up-regulated in the DOP-resupplied cells,indicating diff erential response mechanisms of P.donghaiense to DIP or DOP resupplementation.Our results indicated that P.donghaiense initiated multiple strategies in response to an ambient inorganic P-defi ciency,and its efficient DOP assimilation by providing both P and carbon sources might be a key factor driving bloom formations of P.donghaiense in a low DIP environment.展开更多
Prorocentrum donghaiense,one of the dinoflagellate,has continuously caused large scale red tides along the Chinese coast in recent years. These red tides have brought tremendous loss to the local economy and serious i...Prorocentrum donghaiense,one of the dinoflagellate,has continuously caused large scale red tides along the Chinese coast in recent years. These red tides have brought tremendous loss to the local economy and serious impacts to the local environment. Unfortunately,little was known about the mechanism of the fast appearance and extinction of the red tide caused by this alga. In this study,the full-length cDNA of a caspase encoding gene of P. donghaiense was cloned and characterized,and the transcription of this gene during the senescence of the alga was semi-quantitatively determined. The cDNA was 520 bp in length. It contained a 258 bp open reading frame(ORF) which encoded a peptide of 85 amino acids. The amount of transcripts of the caspase encoding gene increased with the senescence of P. donghaiense and started to decrease gradually when the autolysis of P. donghaiense cells took place. We proposed that the death of P. donghaiense may be a caspase mediated programmed autolysis.展开更多
The effects of different phosphorus(P) concentrations(0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen(N)/P ratios on the growth and photosynthetic ...The effects of different phosphorus(P) concentrations(0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen(N)/P ratios on the growth and photosynthetic characteristics of S keletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P(HP) concentration group showed the greatest algal density and highest specifi c growth rate. Changes in the maximum effi ciency of photosystem Ⅱ(F _v/F_m) were monitored under the various P and N/P conditions. The largest decrease in F _v/F_m was in the low-P(LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields(Φ _(PSⅡ)) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinofl agellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _(PSⅡ) values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang(Yangtze) River estuary from a photosynthesis perspective.展开更多
Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellat...Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs.展开更多
Red tides are a major public hazard in the global oceans.The coast of the East China Sea is the sea area where red tide disasters are the most frequent and serious in China.In order to accurately grasp the occurrence ...Red tides are a major public hazard in the global oceans.The coast of the East China Sea is the sea area where red tide disasters are the most frequent and serious in China.In order to accurately grasp the occurrence of red tides in the coastal waters of the East China Sea,and to understand the microbial communities in the waters during the occurrence of red tides in the East China Sea,a special survey of red tides in the coastal waters of Zhejiang,China was carried out in June 2018.The results showed that nutrient concentrations of N and P were generally high in this area,DIN concentrations in most areas exceeded the permitted limit of Chinese seawater quality grade I.There were significant differences in dissolved oxygen,pH,COD,chlorophyll and phytoplankton abundance of red tides.During the investigation,red tides were found in the waters near the Yushan Islands.The content of chlorophyll a was 42.12mg/m3,the cell abundance of phytoplankton was 8.16×108/L,and the abundance of Prorocentrum edulis accounted for 98.5%.The Illumina MiSeq sequencing platform was used for 16s high-throughput sequencing of water microorganisms,and a total of 16 bacteria were identified.Proteobacteria is the first dominant phylum,followed by Cyanobacteria and Bacteroides.Some differences in bacterial community compositions between HAB and the nearby seawater were observed.The predominant bacteria in the red tide occurrence area were Proteobacteria,comprising 46.1%of the relative abundance;while the predominant bacteria in the nearby sea area,comprising 42.0%of the relative abundance.展开更多
The dinoflagellates Alexandrium pacificum and Prorocentrum donghaiense are two well-known harmful algal blooms(HABs)-forming species,both were usually found in the same sea areas in form of HABs in China.To date,there...The dinoflagellates Alexandrium pacificum and Prorocentrum donghaiense are two well-known harmful algal blooms(HABs)-forming species,both were usually found in the same sea areas in form of HABs in China.To date,there is no mechanistic model that can fully explain the occurrence of P.donghaiense blooms with A.pacificum.We found that different strains of P.donghaiense had different intrinsic growth rates of 0.107-0.215/d,and these strains exhibited different responses to the allelopathic action from A.pacificum.Some strains of P.donghaiense could grow well despite some degrees of inhibition in a short period,suggesting the two algal species P.donghaiense and A.pacificum could coexist,even if A.pacificum was allelopathic.Our findings may advance the understanding of phenotypes in P.donghaiense and provide a potential mechanism involved in the coexistence of P.donghaiense and A.pacificum in the same area.展开更多
PCNA has been considered as a useful marker for the estimation of growth rates of marine phytoplankton at the species level. Since dinoflagellates are noted for having many prokaryotic features in that they are the on...PCNA has been considered as a useful marker for the estimation of growth rates of marine phytoplankton at the species level. Since dinoflagellates are noted for having many prokaryotic features in that they are the only eukaryotes to have permanently condensed chromosomes as well as lacking histones and a nucleosome, the sensitivity to UVB radiation and the validity of PCNA as a maker of growth rate in dinoflagellate need to be evaluated. Prorocentrum donghaiense Lu was investigated to valuate the UVB sensitivity in relation to cellular and molecular aspects of PCNA as a growth indicator. The effects of UVB radiation on PCNA were studied using the methods of western blots technology and whole-cell immunoflurescence with one mono-antibody. UVB changed the cell morphology, halted the growth and increased the cell size, even caused cell death to a certain extent after treatment with UVB radiation in P. donghaiense. Compared with the control, treating the algal cultures in exponential phases with UVB radiation for 24 h caused chromatin release and increases in protein levels of PCNA.展开更多
Prorocentrum donghaiense has caused large-scale red tides off the Chinese coast in recent years. Expressed sequence tag （EST） analysis was carried out for this dinoflagellate in order to identify the functional gene...Prorocentrum donghaiense has caused large-scale red tides off the Chinese coast in recent years. Expressed sequence tag （EST） analysis was carried out for this dinoflagellate in order to identify the functional genes involved in its biological processes. A cDNA library was constructed for P. donghaiense at exponential growth phase, and 565 usable sequencing reads were obtained from 700 clones selected randomly. Messenger RNA corresponding reads were clustered into 36 contigs and 272 singletons （EST groups）. Twenty-two EST groups were found to tag the genes involved in diverse biological processes including programmed cell death （PCD）. Two EST groups showed significant homologies with the encoding genes of cysteine protease （caspase） and proliferating cell nuclear antigen, respectively, two key proteins involved in PCD.展开更多
The response of Prorocentrum donghaiense and Thalassiosira weissfiogii pigments under nitrate （N） and phosphate （P） limitation were studied using HPLC and in vivo fluorescence protocols in batch cultures. For P. d...The response of Prorocentrum donghaiense and Thalassiosira weissfiogii pigments under nitrate （N） and phosphate （P） limitation were studied using HPLC and in vivo fluorescence protocols in batch cultures. For P. donghaiense, the pigment ratio was kept stable under different nutrient conditions from the results of HPLC. For T. weissflogii, there was a lower ratio of chlorophyllide to Chl a during the exponential phase, but the reverse during the stationary phase. Different members of the phytoplankton had different pigments response mechanisms under nutrient limitation. From the results of in vivo fluorescence, the ratio of peridinin to Chl a for P. donghaiense increased in nutrient-free culture, while it was kept stable for nutrient-limited cultures during the exponential phase. For T. weissflogii, the ratio of fucoxanthin to Chl a for each culture increased during the exponential phase, but the ratio under N limitation was apparently lower than that for P limitation during the stationary phase. The results indicate that both pigment ratios from HPLC and in vivo fluorescence of To weissflogii were changed greatly under different nutrient conditions, which suggests that both ratios could be used as indicators of algal physiological status in different nutrient conditions.展开更多
The growth, cellular total lipids, bioaccumulation amount, and bioaccumulation factors （BAFs） of 2,4,4 ′ -tribromodiphenyl ether （BDE28）, 2,2 ′ ,4,4 ′ -tetrabromodiphenyl ether （BDE47）, and 2,2 ′ ,4,4 ′ ,5-...The growth, cellular total lipids, bioaccumulation amount, and bioaccumulation factors （BAFs） of 2,4,4 ′ -tribromodiphenyl ether （BDE28）, 2,2 ′ ,4,4 ′ -tetrabromodiphenyl ether （BDE47）, and 2,2 ′ ,4,4 ′ ,5-pentabromodiphenyl ether （BDE99） in a semi-continuous culture of Prorocentrum donghaiense were studied in relation to nitrate （0, 128, and 512 μmol/L） and phosphate （0, 8, and 32 μmol/L） concentrations. The BDE28, BDE47, and BDE99 content per cell under 0 μmol N/L were 3.77 × 10 6 , 3.95 × 10 6 , and 4.32 × 10 6 ng/cell, respectively, which were significantly higher than those under 128 and 512 μmol N/L. A nearly 5-fold increase in polybrominated diphenyl ether （PBDE） content per algal cell was found between 0 and 8 μmol P/L and between 8 and 32 μmol P/L. With increasing N and P concentrations, the PBDE content per volume of algal culture and the accumulation percentage of available PBDEs declined slightly. The BAFs for the PBDEs based on lipids showed that the logBAF lip under 0 μmol N/L was higher than those under 128 and 512 μmol N/L. The logBAF lip under 0 μmol P/L was higher than that under 8 μmol P/L but lower than that under 32 μmol P/L. Correlation analysis indicated a significant negative correlation between nutrient concentration and cellular total lipids, as well as the PBDE content per cell. The results indicate that different N and P concentrations change the total lipids content of P. donghaiense, thereby resulting in varying PBDE accumulation.展开更多
Modified clay(MC),an effective material used for the emergency elimination of algal blooms,can rapidly reduce the biomass of harmful algal blooms(HABs)via flocculation.After that,MC can still control bloom population ...Modified clay(MC),an effective material used for the emergency elimination of algal blooms,can rapidly reduce the biomass of harmful algal blooms(HABs)via flocculation.After that,MC can still control bloom population through indirect effects such as oxidative stress,whichwas initially proposed to be related to programmedcell death(PCD)at molecular level.To further study theMC induced cell death in residual bloom organisms,especially identifying PCD process,we studied the physiological state of the residual Prorocentrum donghaiense.The experimental results showed that flocculation changed the physiological state of the residual cells,as evidenced by growth inhibition and increased reactive oxygen species production.Moreover,this research provides biochemical and ultrastructural evidence showing that MC induces PCD in P.donghaiense.Nuclear changes were observed,and increased caspase-like activity,externalization of phosphatidylserine and DNA fragmentation were detected in MC-treated groups and quantified.And the mitochondrial apoptosis pathway was activated in both MC-treated groups.Besides,the features of MC-induced PCD in a unicellular organism were summarized and its concentration dependent manner was proved.All our preliminary results elucidate the mechanism through which MC can further control HABs by inducing PCD and suggest a promising application of PCD in bloom control.展开更多
基金The National Natural Science Foundation of China under contract No.42130403the Fundamental Research Funds for the Central Universities under contract No.202362003the National Key Research&Development Program of China under contract No.2017YFC1404300。
文摘Understan ding the mechanism of harmful algal bloom formation is vital for effectively preventing algal bloom outbreaks in coastal environments.Karenia spp.blooms in the East China Sea show a significant correlation with nutrient regimes.However,the impact of key components of nutrients,especially dissolved organic nitrogen(DON),on the blooms of Karenia spp.is not clear.Quantitative research is still lacking.In this study,the cruise observations,field mesocosm-flask culture experiments,and a multinitrogen-tri-phytoplankton-detritus model(NTPD) are combined to reveal the quantitative influence of nutrient regimes on the shift of Prorocentrum donghaiense and Karenia spp.in the East China Sea.It has a synchronism rhythm of diatom-P.donghaienseKarenia spp.-diatom loop in the field culture experiment,which is consistent with the results of the cruise observation.The results showed that the processes of terrigenous DON(TeDON) and dissolved inorganic nitrogen(DIN:NO_(3)^(-)-N,NH_(4)^(+)-N) absorption promoted P.donghaiense to become the dominant algae in the community;whereas the processes of DON from P.donghaiense absorption promoted Karenia spp.to become the dominant algae in ambient DIN exhaustion.In addition,the three-dimensional fluorescence components of humus C,tyrosine and fulvic acid can indicate the processes of growth and extinction of P.donghaiense and Karenia spp.,respectively.This study infers that P.donghaiense and Karenia spp.regime shift mechanism associated with the nutrient regime in coastal waters,which provides a scientific basis for the environmental management of coastal eco system health.
基金The National Basic Research Program of China (973 Program) under contract No.2013CB430302the National High Technology Research and Development Program of China (863 Program) under contract No.2007AA092002+2 种基金the National Natural Science Foundation of China under contract No.41206170the public science and technology research funds projects of the ocean under contract No.201005030scientific research fund of the Second Institute of Oceanography,SOA under contract No.JG1212
文摘Prorocentrum donghaiense is one of the most common red tide causative dinoflagellates in the Changjiang （Yangtze） River Estuary and the adjacent area of the East China Sea. It causes large-scale blooms in late spring and early summer that lead to widespread ecologic and economic damage. A means for distinguish- ing dinoflagellate blooms from diatom （Skeletonema costatum） blooms is desired. On the basis of measure- ments of remote sensing refectance [Rrs（λ）] and inherent optical parameters, the potential of using a mul- tispectral approach is assessed for discriminating the algal blooms due to P. donghaiense from those due to S. costatum. The behavior of two reflectance ratios [R1 = Rrs（560）/Rrs（532） and Re = Rrs（708）/Rrs（665）], suggests that differentiation of P. donghaiense blooms from diatom bloom types is possible from the current band setup of ocean color sensors. It is found that there are two reflectance ratio regimes that indicate a bloom is dominated by P. donghaiense; （1） R1 〉 1.55 and R2 〈 1.0 or （2） R1 〉 1.75 and R2 ≥ 1.0. Various sensitivity analyses are conducted to investigate the effects of the variation in varying levels of chlorophyll concentration and colored dissolved organic matter （CDOM） as well as changes in the backscattering ratio （bbp/bp） on the efficacy of this muitispectral approach. Results indicate that the intensity and inherent op- tical properties of the algal species explain much of the behavior of the two ratios. Although backscattering influences the amplitude of Rrs（λ）, especially in the 530 and 560 nm bands, the discrimination between P. donghaiense and diatoms is not significantly affected by the variation of bbp/bp. Since aCDOM （440） in coastal areas of the ECS is typically lower than 1.0 m-1 in most situations, the presence of CDOM does not interfere with this discrimination, even as SCDOM varies from 0.01 to 0.026 nm-1. Despite all of these effects, the dis- crimination of P. donghaiense blooms from diatom blooms based on multispectral measurements of Rrs（λ） is feasible.
基金Supported by the High Technology Research and Development Program of China (863 Program) (Nos 2006AA09Z180 and 2004AA639790)the National Natural Science Foundation of China (No 40106013)the National Basic Research Program of China (973 Program) (No 2001CB409703)
文摘Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrurn donghaiense at the exponential growth, stationary and decline stages into 〈0.45 μm filtrate, 100 kDa-0.45 μm, 1%100 kDa and 1-10 kDa retentate and 〈1 kDa ultrafiltrate fractions. The fluorescence properties of different molecular weights of dissolved organic matter （DOM） were measured by excitation-emission matrix spectra. Protein-like and humic-like fluorophores were observed in the DOM produced by P. donghaiense. The central positions of protein-like fluorophores showed a red shift with prolonged growth duration, shifting from tyrosine-like properties at the exponential growth stage to tryptophan-like properties at the stationary and decline stages. The excitation wavelengths of protein-like fluorophores exhibited some change in the exponential growth and stationary stages with increased molecular size, but showed little change in the decline stage. However, the emission wavelengths in the decline stage exhibited a blue shift. Very distinct C type and A type peaks in humic-like fluorophores were observed. With a prolonged culture time, the intensities of both of the peaks became strong and the excitation wavelengths of peak A showed a red shift, while the A：C ratios fell. More than 94% of fluorescent DOM was in the lower than 1 kDa molecular weight fraction.
基金The National Natural Science Foundation of China under contract Nos 61533011 and 41776125the NSFCShandong Joint Fund for Marine Ecology and Environmental Sciences under contract No.U1606404+3 种基金the Scientific and Technological Innovation Project of the Qingdao National Laboratory for Marine Science and Technology under contract No.2016ASKJ02the National Key R&D Program of China under contract No.2017YFC1404300the Creative Team Project of the Laboratory for Marine Ecology and Environmental ScienceQingdao National Laboratory for Marine Science and Technology under contract No.LMEESCTSP-2018-1
文摘Most of reported harmful algal blooms(HABs)of microalgae(75%)have been caused by dinoflagellates.Studies on the negative effects of HABs have generally focused on animals,valuable organisms in particular,and environmental factors such as dissolved oxygen and nutrients,but relatively fewer on community level,particularly that using metagenomic approach.In this study,we reported an investigation on the effects of a HAB caused by the dinoflagellate Prorocentrum donghaiense on the species diversity and community structure of the dinoflagellate sub-community via a pyrosequencing approach for the samples taken before,during,and after the bloom season of P.donghaiense in the East China Sea.We sequenced partial 28S rRNA gene of dinoflagellates for the field samples and evaluated the species richness and diversity indices of the dinoflagellate community,as a sub-community of the total phytoplankton.We obtained 800185 valid sequences(categorized into 560 operational taxonomic units,OTUs)of dinoflagellates from 50 samples and found that the biodiversity of dinoflagellate community was significantly reduced during the blooming period in comparison to that in pre-and after-blooming periods,as reflected in the four diversity indices:the species richness expressed as the number of OTUs,Chao1 index,Shannon index(evenness),and Gini-Simpson index.These four indices were all found to be negatively correlated to the cell density of the bloom species P.donghaiense.Correlation analyses also revealed that the P.donghaiense cell abundance was correlated negatively with NO3--N,and NO2--N,but positively with total nitrogen(TN)and total phosphorus(TP).Principal coordinates analysis(PCoA)showed that the community structure of dinoflagellates was markedly different among the different sampling periods,while the redundancy analysis(RDA)revealed P.donghaiense abundance,salinity,NO3--N,and SiO32-were the most four significant factors shaping the dinoflagellate community structure.Our results together demonstrated that HABs caused by the dinoflagellate P.donghaiense could strongly impact the aquatic ecosystem on the sub-community level which the blooming species belongs to.
基金Supported by the National Natural Science Foundation of China (Nos.40406028, 40706044)
文摘Mitochondrial cytochrome b （Cytb）, one of the few proteins encoded by the mitochondrial DNA, plays an important role in transferring electrons. As a mitochondrial gene, it has been widely used for phylogenetic analysis. Previously, a 949-bp fragment of the coding gene and mRNA editing were characterized from Prorocentrum donghaiense, which might prove useful for resolving P. donghaiense from closely related species. However, the full-length coding region has not been characterized. Ih this study, we used rapid amplification of cDNA ends （RACE） to obtain full-length, 1 124 bp cDNA. Cytb transcript contained a standard initiation codon ATG, but did not have a recognizable stop codon. Homology comparison showed that the P. donghaiense Cytb had a high sequence identity to Cytb sequences from other dinoflagellate species. Phylogenetic analysis placed Cytb from P. donghaiense in the clade of dinoflagellates and it clustered together strongly with that from P. minimum. Based on the full-length sequence, we inferred 32 editing events at different positions, accounting for 2.93% of the Cytb gene. 34.4% （11） of the changes were A to G, 25% （8） were T to C, and 25% （8） were C to U, with smaller proportions of G to C and G to A edits （9.4% （3） and 6.2% （2）, respectively）. The expression level of the Cytb transcript was quantified by real-time PCR with a TaqMan probe at different times during the whole growth phase. The average Cytb transcript was present at 39.277.46 copies of cDNA per cell during the whole growth cycle, and the expression of Cytb was relatively stable over the different phases. These results deepen our understanding of the structure and characteristics of Cytb in P. donghaiense, and confirmed that Cytb in P. donghaiense is a candidate reference gene for studying the expression of other genes.
基金The National Natural Science Foundation of China(NSFC)under contract Nos 41276186,41506015 and 41606038the NSFC-Shandong Joint Fund for Marine Science Research Centers under contract No.U1606405the Postdoctoral Innovation Foundation of Shandong Province under contract No.201502031
文摘Algal blooms caused by Prorocentrum donghaiense occurred frequently in the East China Sea （ECS） during spring in recent years. In this study, a coupled biophysical model was used to hindcast a massive P. donghaiense bloom that occurred in 2005 and to determine the factors influencing bloom initiation and development. The model comprised the Regional Ocean Modeling System tailored for the ECS that utilized a multi-nested configuration and a population dynamics model for 19. donghaiense. Comparisons between simulations and observations revealed that the biological model is capable of reproducing the characteristics of 19. donghaiense growth under different irradiances and phosphorus limitation scenarios. The variation of intracellular phosphorus and the effects of 19. donghaiense on ambient nutrients conditions were also reproduced. The biophysical model hindcasted the hydrodynamics and spatiotemporal distributions of the P. donghaiense bloom reasonably well. Bloom development was consistent with observations reported in earlier studies. The results demonstrate the capability of the model in capturing subsurface incubation during bloom initiation. Then model＇s hindcast solutions were further used to diagnose the factors controlling the vertical distribution. Phosphate appeared to be one of the factors controlling the subsurface incubation, whereas surface wind fields played an important role in determining P. donghaiense distribution. The results highlight the importance of nutrient-limitation as a mechanism in the formation of P. donghaiense subsurface layers and the dispersing of P. donghaiense blooms. This coupled biophysical model should be improved and used to investigate 19. donghaiense blooms occurring in different scenarios.
基金The National High Technology Research and Development Program ("863" Program) of China under contract No 2006AA09Z178 the National Natural Science Foundation of China under contract No 40706044
文摘Prorocentrum donghaiense is an important harmful algae bloom （HAB） causing creature in China＇s seas, and the conventional visual detection can not cope with long-term monitoring and highthroughput sampling projects. An assay for P. donghaiense with sandwich hybridization integrated with nuclease protection assay （NPA-SH） was established. Tests with mixed samples and spiked field ones confirmed its good specificity and sensitivity. The cell number of P. donghaiense correlated well with the optical density, and the regression equation is y=4× 10^- 6x＋ 0.694 9, in which x is the cell number, and y is the optical density, with r2=0.953 5. These results show that the NPA-SH method has good feasibility in the detection of P. donghaiense. Results of NPA-SH and microscopy are excellent for each sample. The NPA-SH method was a simple way in quantitative detection of P. donghaiense, and the whole process could be finished in about six hours, which provided a new approach in high-throughput sampling and long-term monitoring of P. donghaiense.
基金Supported by the National Key R&D Program of China(No.2018YFD0901500)the National Natural Science Foundation(Nos.41876124,61871293,41706147,42007372)。
文摘Harmful algal bloom(HAB)is an ecological disaster to local mariculture.At present,its impact on macrophytes has not been well studied.In this study,we cultivated sexually propagated embryos of S argassum fusiformis-an edible seaweed-in Prorocentrum donghaiense suspensions at different cell densities(0,0.50×10^(5),0.75×10^(5),1.00×10^(5),and 1.50×10^(5) cells/mL)for 10 days,during which growth and photosynthetic activities of the embryos were determined,and a monocultivation was set up for comparison.Results show that the relative growth rate and photosynthetic activities of the embryos co-cultivated with P.donghaiense were inhibited mostly and significantly in the cell densities of 0.75×10^(5),1.00×10^(5),and 1.50×10^(5) cells/mL,and the inhibitory effects increased in overall with increased cell densities.The maximum relative electron transport rates(rETR max)and apparent photosynthetic efficiency(a)of co-cultivated embryos were all significantly lower than monocultivation ones on the 10 th day.Furthermore,the photosynthetic activity detected by chlorophyll-a fluorescence transient(i.e.,OJIP),the electron transport among electron transfer accepters of PSII(photosystem II)and that from PSII to PSI(photosystem I)was restricted,which is probably responsible for the decreases of rETR max andain the co-cultivated embryos.In addition,parts of the photosynthetic reaction centers of PSII in the co-cultivated embryos were inactivated.Therefore,P.donghaiense bloom could restrain the development and photosynthetic activities of S.fusiformis embryos,reduce the seedlings stock,and eventually hinder the development of S.fusiformis production industry.
基金supported by the National Natural Science Foundation of China (41076065)the Major State Basic Research Development Program of China (2010CB428701)
文摘Interactions between Skeletonema costatum (S. costatum) and Prorocentrum donghaiense (P. donghaiiense) were inves-tigated using bi-algal cultures at different concentrations of phosphate (PO4-P) and nitrate/phosphate (N/P) ratios. Experiments were conducted under P-limited conditions and the Lotka-Volterra mathematical model was used to simulate the growth of S. costatum and P. donghaiense in the bi-algal cultures. Both of these two species were inhibited significantly in bi-algal culture. The results of the simulation showed that the inhibitory degree of S. costaum by P. donghaiense was high when the concentration of PO4-P was low (0.1μmolL-1/2 d), but that of P. donghaiense by S. costaum was high with increased PO4-P supply (0.6μmolL-1/2 d). At low concen-tration of PO4-P (0.1μmolL-1/2 d), or high concentration of PO4-P (0.6μmolL-1/2 d) with high N/P ratio (160), the interactions be-tween S. costatum and P. donghaiense were dependent on the initial cell densities of both species. At high concentration of PO4-P (0.6μmolL-1/2 d) with low N/P ratio (25 or 80), S. costatum exhibited a survival strategy superior to that of P. donghaiense. The de-gree of inhibition of P. donghaiense by S. costaum increased with elevated N/P ratio when the medium was supplemented with con-centration 0.1μmolL-1/2 d of PO4-P. The degree of inhibition to P. donghaiense by S. costaum increased with elevated N/P ratio at low concentration of PO4-P (0.1 μmolL-1/2 d). This trend was conversed at high concentration of PO4-P (0.6μmolL-1/2 d). However, the degree of inhibition of S. costaum by P. donghaiense increased with the increased N/P ratio at different PO4-P concentrations (0.1μmolL-1/2 d and 0.6μmolL-1/2 d). These results suggested that both phosphate concentration and N/P ratio affected the competition between S. costaum and P. donghaiense: P. donghaiense is more competitive in environments with low phosphate or high N/P ratio and the influence of N/P ratio on the competition was more significant with lower phosphate concentration.
基金Supported by the National Key Research Development Program of China(No.2017YFC1404302)the National Natural Science Foundation of China(Nos.41425021,41706131)+1 种基金the Open Fund of CAS Key Laboratory of Marine Ecology and Environmental Sciences,Institute of Oceanology,Chinese Academy of Sciences(No.KLMEES201806)supported by the“Ten-Thousand Talents Program”for leading talents in science and technological innovation。
文摘Dinofl agellates are the major causative agents of harmful algal blooms in the global ocean and they usually form blooms under conditions of very low dissolved inorganic phosphorus(DIP).However,the mechanisms underpinning the dinofl agellate blooms remain unclear.Here,we quantitatively compared protein expression profi les of a marine dinofl agellate,Prorocentrum donghaiense,grown in inorganic P-replete,P-defi cient,and DIP-and dissolved organic phosphorus(DOP)-resupplied conditions by employing a Tandem Mass Tag(TMT)-based quantitative proteomic approach.Proteins involved in intracellular P reallocation,organic P,and non-P lipid utilization were up-regulated under the P-defi cient condition,while inorganic phosphate transporters varied insignifi cantly.In response to the P resupplementation,nitrogen metabolism,ribosome,porphyrin,and chlorophyll metabolism were up-regulated,while lysosome,and starch and sucrose metabolism were down-regulated.Notably,photosynthesis was up-regulated and secondary metabolism was down-regulated only in the DIP-resupplied cells,whereas amino acid metabolism and vitamin B6 metabolism were up-regulated in the DOP-resupplied cells,indicating diff erential response mechanisms of P.donghaiense to DIP or DOP resupplementation.Our results indicated that P.donghaiense initiated multiple strategies in response to an ambient inorganic P-defi ciency,and its efficient DOP assimilation by providing both P and carbon sources might be a key factor driving bloom formations of P.donghaiense in a low DIP environment.
文摘Prorocentrum donghaiense,one of the dinoflagellate,has continuously caused large scale red tides along the Chinese coast in recent years. These red tides have brought tremendous loss to the local economy and serious impacts to the local environment. Unfortunately,little was known about the mechanism of the fast appearance and extinction of the red tide caused by this alga. In this study,the full-length cDNA of a caspase encoding gene of P. donghaiense was cloned and characterized,and the transcription of this gene during the senescence of the alga was semi-quantitatively determined. The cDNA was 520 bp in length. It contained a 258 bp open reading frame(ORF) which encoded a peptide of 85 amino acids. The amount of transcripts of the caspase encoding gene increased with the senescence of P. donghaiense and started to decrease gradually when the autolysis of P. donghaiense cells took place. We proposed that the death of P. donghaiense may be a caspase mediated programmed autolysis.
基金Supported by the“Strategic Priority Research Program-Climate Change:Carbon Budget and Relevant Issues”of Chinese Academy of Sciences(No.XDA05030401)the National Basic Research Program of China(973 Program)(No.2014CB441504)+1 种基金the Key Program of National Natural Science Foundation of China(No.41230963)the NSFCShandong Province Joint Fund Project(No.U1406403)
文摘The effects of different phosphorus(P) concentrations(0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen(N)/P ratios on the growth and photosynthetic characteristics of S keletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P(HP) concentration group showed the greatest algal density and highest specifi c growth rate. Changes in the maximum effi ciency of photosystem Ⅱ(F _v/F_m) were monitored under the various P and N/P conditions. The largest decrease in F _v/F_m was in the low-P(LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields(Φ _(PSⅡ)) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinofl agellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _(PSⅡ) values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang(Yangtze) River estuary from a photosynthesis perspective.
基金supported by the National Basic Research Program of China (973 Program) (No.2011CB403602)the National High Technology Research and Development Program of China (863 Program) (No.2007AA09200111)the National Marine Public Welfare Research Project (201205031-02)
文摘Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs.
基金National Key Research and Development Program“High-resolution Numerical Simulation and Prediction of Ecological Environment in the Yellow and East China Seas”(Project No.:2016YFC1401603)Scientific Research Project of Department of Ecology and Environment of Zhejiang Province(Project No.:2016A012).
文摘Red tides are a major public hazard in the global oceans.The coast of the East China Sea is the sea area where red tide disasters are the most frequent and serious in China.In order to accurately grasp the occurrence of red tides in the coastal waters of the East China Sea,and to understand the microbial communities in the waters during the occurrence of red tides in the East China Sea,a special survey of red tides in the coastal waters of Zhejiang,China was carried out in June 2018.The results showed that nutrient concentrations of N and P were generally high in this area,DIN concentrations in most areas exceeded the permitted limit of Chinese seawater quality grade I.There were significant differences in dissolved oxygen,pH,COD,chlorophyll and phytoplankton abundance of red tides.During the investigation,red tides were found in the waters near the Yushan Islands.The content of chlorophyll a was 42.12mg/m3,the cell abundance of phytoplankton was 8.16×108/L,and the abundance of Prorocentrum edulis accounted for 98.5%.The Illumina MiSeq sequencing platform was used for 16s high-throughput sequencing of water microorganisms,and a total of 16 bacteria were identified.Proteobacteria is the first dominant phylum,followed by Cyanobacteria and Bacteroides.Some differences in bacterial community compositions between HAB and the nearby seawater were observed.The predominant bacteria in the red tide occurrence area were Proteobacteria,comprising 46.1%of the relative abundance;while the predominant bacteria in the nearby sea area,comprising 42.0%of the relative abundance.
基金Supported by the National Key Research and Development Program of China(No.2017YFC1404301)the National Natural Science Foundation of China(Nos.41230961,41176088)。
文摘The dinoflagellates Alexandrium pacificum and Prorocentrum donghaiense are two well-known harmful algal blooms(HABs)-forming species,both were usually found in the same sea areas in form of HABs in China.To date,there is no mechanistic model that can fully explain the occurrence of P.donghaiense blooms with A.pacificum.We found that different strains of P.donghaiense had different intrinsic growth rates of 0.107-0.215/d,and these strains exhibited different responses to the allelopathic action from A.pacificum.Some strains of P.donghaiense could grow well despite some degrees of inhibition in a short period,suggesting the two algal species P.donghaiense and A.pacificum could coexist,even if A.pacificum was allelopathic.Our findings may advance the understanding of phenotypes in P.donghaiense and provide a potential mechanism involved in the coexistence of P.donghaiense and A.pacificum in the same area.
基金the National Key Basic Research and Development Program （red tide ＂973＂program） of China under contract No. 2301CB409700 the National Natural Science Foundation of China under contract Nos 40232021,405211303 Ministry of Science and Technology of China under contract Nos 2005AA635240 and MEL0608.
文摘PCNA has been considered as a useful marker for the estimation of growth rates of marine phytoplankton at the species level. Since dinoflagellates are noted for having many prokaryotic features in that they are the only eukaryotes to have permanently condensed chromosomes as well as lacking histones and a nucleosome, the sensitivity to UVB radiation and the validity of PCNA as a maker of growth rate in dinoflagellate need to be evaluated. Prorocentrum donghaiense Lu was investigated to valuate the UVB sensitivity in relation to cellular and molecular aspects of PCNA as a growth indicator. The effects of UVB radiation on PCNA were studied using the methods of western blots technology and whole-cell immunoflurescence with one mono-antibody. UVB changed the cell morphology, halted the growth and increased the cell size, even caused cell death to a certain extent after treatment with UVB radiation in P. donghaiense. Compared with the control, treating the algal cultures in exponential phases with UVB radiation for 24 h caused chromatin release and increases in protein levels of PCNA.
基金Supported by the Key Laboratory of Mariculture of Chinese Education Ministry, Ocean University of China (No. 200405)the Ministry of Science and Technology of China (No. 2005CCA02400,2001CB409700)
文摘Prorocentrum donghaiense has caused large-scale red tides off the Chinese coast in recent years. Expressed sequence tag （EST） analysis was carried out for this dinoflagellate in order to identify the functional genes involved in its biological processes. A cDNA library was constructed for P. donghaiense at exponential growth phase, and 565 usable sequencing reads were obtained from 700 clones selected randomly. Messenger RNA corresponding reads were clustered into 36 contigs and 272 singletons （EST groups）. Twenty-two EST groups were found to tag the genes involved in diverse biological processes including programmed cell death （PCD）. Two EST groups showed significant homologies with the encoding genes of cysteine protease （caspase） and proliferating cell nuclear antigen, respectively, two key proteins involved in PCD.
基金Supported by the National Natural Science Foundation of China （40076031 40376043）, the China Postdoctoral Science Foundation （20060400854, to J. J. Hou） and the National Basic Research Programme （2001CB409704）.The authors would like to thank Mr Xiang-Hui Dai, Hao Zhang, Jia-Bing Lin and Wei-Min Yuan in Environmental Science Research Center, Xiamen University for their assistance during this study. Prof. Ivor John Hodgkiss in Department of Ecology and Biodiversity, University of Hong Kong is thanked for carefully reviewing the manuscript and giving many constructive suggestions.
文摘The response of Prorocentrum donghaiense and Thalassiosira weissfiogii pigments under nitrate （N） and phosphate （P） limitation were studied using HPLC and in vivo fluorescence protocols in batch cultures. For P. donghaiense, the pigment ratio was kept stable under different nutrient conditions from the results of HPLC. For T. weissflogii, there was a lower ratio of chlorophyllide to Chl a during the exponential phase, but the reverse during the stationary phase. Different members of the phytoplankton had different pigments response mechanisms under nutrient limitation. From the results of in vivo fluorescence, the ratio of peridinin to Chl a for P. donghaiense increased in nutrient-free culture, while it was kept stable for nutrient-limited cultures during the exponential phase. For T. weissflogii, the ratio of fucoxanthin to Chl a for each culture increased during the exponential phase, but the ratio under N limitation was apparently lower than that for P limitation during the stationary phase. The results indicate that both pigment ratios from HPLC and in vivo fluorescence of To weissflogii were changed greatly under different nutrient conditions, which suggests that both ratios could be used as indicators of algal physiological status in different nutrient conditions.
基金supported by the National Natural Science Foundation of China (No.40906061)the "Two Districts" Foundation of Shandong Province, China (No.2011-Yellow-19)+1 种基金the Science and Technology Plan Projects of Qingdao (No.12-1-3-64-nsh)the Talent Foundation of Qingdao Agricultural University (No.630642)
文摘The growth, cellular total lipids, bioaccumulation amount, and bioaccumulation factors （BAFs） of 2,4,4 ′ -tribromodiphenyl ether （BDE28）, 2,2 ′ ,4,4 ′ -tetrabromodiphenyl ether （BDE47）, and 2,2 ′ ,4,4 ′ ,5-pentabromodiphenyl ether （BDE99） in a semi-continuous culture of Prorocentrum donghaiense were studied in relation to nitrate （0, 128, and 512 μmol/L） and phosphate （0, 8, and 32 μmol/L） concentrations. The BDE28, BDE47, and BDE99 content per cell under 0 μmol N/L were 3.77 × 10 6 , 3.95 × 10 6 , and 4.32 × 10 6 ng/cell, respectively, which were significantly higher than those under 128 and 512 μmol N/L. A nearly 5-fold increase in polybrominated diphenyl ether （PBDE） content per algal cell was found between 0 and 8 μmol P/L and between 8 and 32 μmol P/L. With increasing N and P concentrations, the PBDE content per volume of algal culture and the accumulation percentage of available PBDEs declined slightly. The BAFs for the PBDEs based on lipids showed that the logBAF lip under 0 μmol N/L was higher than those under 128 and 512 μmol N/L. The logBAF lip under 0 μmol P/L was higher than that under 8 μmol P/L but lower than that under 32 μmol P/L. Correlation analysis indicated a significant negative correlation between nutrient concentration and cellular total lipids, as well as the PBDE content per cell. The results indicate that different N and P concentrations change the total lipids content of P. donghaiense, thereby resulting in varying PBDE accumulation.
基金supported by the Taishan Scholars Climbing Program of Shandong Province of 2019,the Special Research Assistant Program of Chinese Academy of Sciences,China Postdoctoral Science Foundation Funded Project(No.2020M672153)the AoShan Talents Cultivation Program from Qingdao National Laboratory for Marine Science and Technology(No.2017ASTCP-OS16).
文摘Modified clay(MC),an effective material used for the emergency elimination of algal blooms,can rapidly reduce the biomass of harmful algal blooms(HABs)via flocculation.After that,MC can still control bloom population through indirect effects such as oxidative stress,whichwas initially proposed to be related to programmedcell death(PCD)at molecular level.To further study theMC induced cell death in residual bloom organisms,especially identifying PCD process,we studied the physiological state of the residual Prorocentrum donghaiense.The experimental results showed that flocculation changed the physiological state of the residual cells,as evidenced by growth inhibition and increased reactive oxygen species production.Moreover,this research provides biochemical and ultrastructural evidence showing that MC induces PCD in P.donghaiense.Nuclear changes were observed,and increased caspase-like activity,externalization of phosphatidylserine and DNA fragmentation were detected in MC-treated groups and quantified.And the mitochondrial apoptosis pathway was activated in both MC-treated groups.Besides,the features of MC-induced PCD in a unicellular organism were summarized and its concentration dependent manner was proved.All our preliminary results elucidate the mechanism through which MC can further control HABs by inducing PCD and suggest a promising application of PCD in bloom control.
