Dinoflagellate Cysts Records from Core Samples of Modern Marine Sediment at the Luoyuan Bay Mouth

Thirty species of dinoflagellate cysts in 15 genus are identified from KMZK5 Core samples of recent marine sediment at the Luoyuan Bay mouth, Fujian. All of these dinoflagellate cysts are first recorded in the Luyuan Bay, 12 species of them are not distributing in the near sea area such as Sansha Bay and Minjiang estuary, including 6 kinds of toxic species, such as Alexandrium affine, A. minutum, A. tamarense, Gonyaulax spinifera, Gymnodinium catenatum and Scrippsiella trochoidea. The abundance and vertical distribution characterestics of the main and the toxic dinoflgellate cysts are also studied in the paper.

Distribution characteristic and variation trend of planktonic dinoflagellate in the Taiwan Strait from 2006 to 2007

Based on 4 cruise surveys from July 2006 to October 2007 in the Taiwan Strait, the species composition, community structure and spatio-temporal distribution of dinoflagellate were studied. A total of 131 dinoflagellates belonging to 18 genera were identified. The population was dominated by hyperthermal and hyperhaline species accounting for 72.52% of the total species. Eurythermal and euryhaline species were the second most common one accounting for 25.19% of the total species. It was only 2.29% for neritic species. The maximum species number occurred in summer, while the maximum cell density appeared in spring. The average dinoflagellate cell density was 404.96x104 cells/m3. It showed that the dinof]agellate cell density increased from the nearshore waters to the open sea and from the north to the south. Compared with the results during 1984-1985, the horizontal distribution pattern and seaeonal variation of the dinofiagellate have not changed significantly, but the dinoflagellate cell density increased by 3.01 times. Further analysis of the dinoflagellate abundance variations both in the spatial and temporal aspects, indicated that the abundance of dinoflagellate increased more significantly in cold seasons, and there was a larger increase in the north of the Taiwan Strait. Besides, the dinoflagellate community structure changed notably. It showed that the diversity and evenness index were relatively high, and the proportion of dinoflageliate cell density to the total phytoplankton increased.

Biological and Molecular Geochemical Evidence for Dinoflagellate Ancestors in the Upper Sinian-Cambrian

Dinoflagellates are single celled organisms that reflect the ecological conditions in modern oceans and lakes. Their earliest undisputed fossil record suggests that dinoflagellates originated from the Middle Triassic (c. 240 Ma ago). However, the presence of molecular biomarkers (dinosterane, 4α-methyl-24-ethylcholestane and tria-romatic dinosteroids) in rock extracts and coccoid dinoflagellate fossils from the upper Sinian to Cambrian of the Tarim basin confirms the hypothesis that dinoflagellates have an ancient origin, and predate the oldest undisputed dinoflagellate fossils at least by 300 Ma, as early as the late Sinian-Cambrian.

Effects of ambient DIN:DIP ratio on the nitrogen uptake of harmful dinoflagellate Prorocentrum minimum and Prorocentrum donghaiense in turbidistat

The effects of varying nitrogen （N）： phosphorus （P） ratios on the growth and N-uptake and assimilation of the harmful dinoflagellates Prorocentrum minimum and Prorocentrum donghaiense were examined in turbidistat culture experiments. Algal cultures were supplied with media containing PO4^3- in various concentrations to obtain a wide range of N：P ratios. Experiments to determine rates of N uptake and assimilation of different N sources （NO^3-, NH4^＋, urea and glycine by P. minimum and NO3^-, NH4^＋ by P. donghaiense） were conducted using ^15-N tracer techniques at each N：P ratio. The growth rates suggested nutrient limitation at both high and low N：P ratios relative to the Redfield ratio. On a diel basis, the growth of both species was regulated by the light-dark cycle, which may be a result of regulation of both lightdependent growth and light-independent nutrient uptake. Maximum growth rates of both species always occurred at the beginning of light phase. In P-rich medium （low N：P ratio）, both species had higher N assimilation rates, suggesting N limitation. Low assimilation coefficients at high N：P ratios suggested P limitation of N uptake and assimilation. NO3 ^-and NH4^＋ contributed more than 90% of the total N uptake of P. minimum. Reduced N sources were more quickly assimilated than NO3^-. Highest average daily growth rates were recorded near an N：P ratio of 12 for both species. The N uptake rates of cultures at N：P ratios near Redfield ratio were more balanced with growth rates. The linkage between growth rates and N uptake/assimilation rates were conceptually described by the variation of cell N quota. The N：P ratios affect the N uptake and growth of Prorocentrum spp., and may regulate their bloom progression in eutrophic ecosystems.

Distribution of dinoflagellate cysts in Yellow Sea sediments

To investigate the distribution, abundance, and species composition of dinoflagellate cysts in the Yellow Sea, surface sediment samples were collected at 37 sites, including the Korean dump site. Twenty-one di- noflagellate cyst taxa were identified, with the assemblages dominated mainly by Spiniferites bulloideus, Operculodinium centrocarpum, and cyst of Alexandrium catenella/tarnarense type. A high frequency of O. centrocarpum in the Yellow Sea was observed for the first time, and it is likely that this can be attributed to the dynamics of the Yellow Sea Cold Water Mass and the Changjiang （Yangtze） River runoff. Total cyst concentrations ranged from 23 to 48 442 cysts/g dry weight, and high cyst concentrations were recorded adjacent to the dumping site. This result suggests that anthropogenic activities such as ocean dumping s- timulate the growth of dinoflagellates in the Yellow Sea, which in turn leads to high levels of dinoflagellate cyst production.

Abundance and distribution of dinoflagellate cysts in Xiamen Western Harbor

In a grid investigation, dinoflagellate cysts were collected from sediments in Xiamen Western Harbor in May of 2000, from which five species of cysts were identified: Alexandrium tamarensis, A. minutum, Lingulodinium polyedra, Gonyaulax scrippsae and Gymnodinium catenatum, account for about 21% in the species composition. The quanti-tative analysis of the sediments shows that the number of dinoflagellate cysts varies from 51 to 256 cysts/g of sediment, the highest value (>200 cysts/g) being recorded at the stations of the central part of the bay, while the lowest (<100 cysts/g) at the bay mouth. A good linear relationship is found between cyst amount and fine-grained sediments. Complex physiognomies on the seabed, topographty in the bay and weak water exchange are the main factors not only in cyst accumulation but also in their distribution pattern, and have resulted in the difference in cyst densities between the inner bay and the outer bay in the harbor.

Dinoflagellate cysts in recent marine sediment from Guangxi, China

Total of 33 species of dinoflagellate cysts were discovered from surface sediment in the sea region of Guangxi, among them 12 cyst types (Diplopsalopsis sp. 1, D. sp. 2, D. sp. 3, Cochlodinium sp., Protoperidinium sp. 1, P. sp. 2, P. compressum, Scrippsiella sp. 1, S. sp. 2, Alexandrium sp. 1, A. sp. 2, A. sp. 3) were first reported from the South China Sea. And one cyst type (Cochlodinium sp.) was first reported in the world. Scrippsiella trochoidea is the dominant species in this area, accounting for 45% of all the cysts. There are 2 cysts of toxic dinoflagellate (Alexandrium tamarensis and Gymnodinium catenatum) But there is no relationship between cyst number and grain size distribution.

Temporal population dynamics of dinoflagellate Prorocentrum minimum in a semi-enclosed mariculture pond and its relationship to environmental factors and protozoan grazers

The ecological processes and interrelationships between protists,either autotrophic or heterotrophic,and environmental factors in mariculture ponds are largely unknown.This study investigated the temporal dynamics of potentially harmful dinoflagellate,Prorocentrum minimum (Pavillard) Schiller,and its relationship to physico-chemical factors and protozoan grazers over a complete cycle in a semi-enclosed shrimp-farming pond near Qingdao,Northern China.P.minimum occurred frequently in low numbers from June to August,followed by a sharp increase from the middle of August,reaching a single maximum peak value of 2.2×105 cells L-1 in October.Temporal variation in abundance was positively correlated with dissolved nitrogen,but showed a significant inverse relationship to abundance of the dominant ciliates,Tintinnopsis lohmanni and Askenasia stellaris.The results provide statistical evidence that the number of P.minimum increased with increasing nitrogen,and the suppression or shortening of algal bloom may be associated with protozoan grazers,such as Tintinnopsis lohmanni,in mariculture ponds.

Blooms of Prorocentrum donghaiense reduced the species diversity of dinoflagellate community

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.

Effects of Ultraviolet(UV)Radiation on Outdoor-and Indoor-Cultured Prorocentrum lima,a Toxic Benthic Dinoflagellate

The effects of different types of ultraviolet(UV)radiation(UVR,wavelength=280-400 nm)and light intensities on cell growth,pigment composition,UV-absorbing compounds(UVACs),and chlorophyll a(Chl a)fluorescence were studied in dinoflag-ellate Prorocentrum lima cultured outdoors for 16 days and indoors for 18 days.In the outdoor experiment,UVA radiation(320-400 nm)increased the growth rate of this dinoflagellate when solar light intensities were<12%;decreased growth rates were observed when intensities were>12%.Exposure to UVB radiation(280-320 nm)alleviated the negative effects of UVA.In the indoor ex-periment,UVA and low doses of UVB enhanced growth rates.Addition of low doses of UVB to UVA exposure resulted in higher contents of Chl a and photoprotective pigments compared with UVA exposure only.The results of both experiments showed that UVB is the primary signal of UVAC synthesis.High-dose UVB exposure accelerated growth rates when UVAC contents were maintained at high levels,suggesting that the latter plays a key role in UVR damage protection.Furthermore,the repair rate was en-hanced by UVB exposure after 16 days of culture.This study confirms the positive effects of UVA and UVB on the growth of P.lima,with the latter enhancing the photoprotective and recovery pathways of the species.

Phylogenetic diversity of dinoflagellates in polar regions

Because of the limitations of sampling and seasonal study in polar regions, knowledge of dinoflagellate diversity, distribution and ecology are limited. Dinoflagellates have been incidentally reported from polar regions during some seasons and some populations have been reported as components of microalgae. Surveys of molecular diversity link the genotype of dinoflagellates from polar regions with environmental adaptation. In this study, 37 positive clones of dinoflagellates collected from different sites were used for genotype analysis, providing new insights into the biodiversity and distribution of these species based on 18S rRNA sequencing. Diverse genotypes were recorded for the summer season in Kongsfjorden （high Arctic） whilst a single novel genotype of dinoflagellate was recorded from winter samples from the Antarctic Ocean. Data from ice cores suggests that this single dinoflagellate genotype was adapted to extreme cold and clone library screening found that it was occasionally the only microbial eukaryotic genotype found in winter ice cores. The findings of this study could improve our understanding of the diverse dinoflagellate genotypes occurring in these perennially cold microbial ecosystems.

Antialgal and Antilarval Activities of Bioactive Compounds Extracted from the Marine Dinoflagellate Amphidinium carterae

Abstract With the global ban on the application of organotin-based marine coatings by the International Maritime Organization, the development of environmentally friendly, low-toxic and nontoxic antifouling compounds for marine industries has become an urgent need. Marine microorganisms have been considered as a potential source of natural antifoulants. In this study, the antifouling potential of marine dinoflagellate Amphidinium carterae, the toxic and red-tide microalgae, was investigated. We performed a series of operations to extract the bioactive substances from Amphidinium carterae and tested their antialgal and antilarval activities. The crude extract of Amphidinium carterae showed significant antialgal activity and the ECs0 value against Skeletonema costatum was 55.4 ~tg mL-1. After purification, the isolated bioactive substances （the organic extract C） exhibited much higher antialgal and antilar- val activities with ECso of 12.9μgmL-1 against Skeletonema costatum and LC50 of 15.1 μmL-1 against Amphibalanus amphitrite larvae. Subsequently, IR, Q-TOFMS, and GC-MS were utilized for the structural elucidation of the bioactive compounds, and a series of unsaturated and saturated 16- to 22-carbon fatty acids were detected. The data suggested the bioactive compounds isolated from Amphidinium carterae exhibited a significant inhibiting effect against the diatom Skeletonema costatum and Amphibalanus amphi- trite larvae, and could be substitutes for persistent, toxic antifouling compounds.

Preliminary study on taxonomy of dinoflagellate cysts from major estuary and bays of Fujian Province, China

According to the morphology, wall structure, color, ornamentation, etc., 25 species belonging to 9 genera are identified and described from 144 sediment samples of the Xiamen Harbor, the mouth of the Minjiang River and the Sansha Bay. Among them there are 2 toxic species: Alexandrium minutun,A. tamarenes, 4 harmful species: Alexandrium affine,Lingulodinium polX drum, Scrippsiella trochoide,Gonyaulax spinifera. It shows that 11 species of dinoflagellate cysts (Alexandrium affine, A. minutum, Diplopelta cf. parva, Polykrikos cf. schwartzii, protoceratium reticulatum, Protoperidinium minutum, P. cf. minutum, P. cf americanum and Alexandrium sp., Protoperidinium sp. 1, P. sp. 2) are first recorded along the coast of Fujian Province, China. These newly discovered species might be transported to the coastal sea of Fujian Province by ballast water of international trade vessels.

Co-monitoring bacterial and dinoflagellates communities by denaturing gradient gel electrophoresis (DGGE) and SSU rRNA sequencing during a dinoflagellates bloom

Dinoflagellates are unicellular eukaryotic protists that dominate in all coastal waters, and are also present in oceanic waters. Despite the central importance of dinoflagellates in global primary production, the relationship between dinoflagellates and bacteria are still poorly understood. In order to understand the ecological interaction between bacterial and dinoflagellates communities, denaturing gradient gel electrophoresis (DGGE) and SSU rRNA sequencing were applied to monitoring the population dynamics of bacteria and dinoflagellates from the onset to disappearance of a dinoflagellates bloom occurred in Baltimore Inner Harbor, from April 15 to 24,2002. Although Prorocentrum minimum was the major bloom forming species under the light microscopy, DGGE method with dinoflagellate specific primers demonstrated that Prorocentrum micans, Gymnodinium galatheanum and Gyrodinium uncatenum were also present during the bloom. Population shifts among the minor dinoflagellate groups were observed. DGGE of PCR-amplified 16S rRNA gene fragments indicated that cyanobacteria, α, β, γ-proteobacteria, Flavobacterium Bacteroides-Cytophaga (FBC), and Planctomcetes were the major components of bacterial assemblages during the bloom. DGGE analysis showed that Cytophagales and α-proteobacteria played important roles at different stages of dinoflagellates bloom. DGGE can be used as a rapid tool to simultaneously monitor population dynamics of both bacterial and dinoflagellates communities in aquatic environments, which is demonstrated here.

Biodiversity and distribution of benthic dinoflagellates in tropical Zhongsha Islands,South China Sea

Benthic dinoflagellates have attracted increasing attention in recent years because of their toxicity and ability to form extensive harmful algal blooms.Ostreopsis producing palytoxin and its analogs,Gambierdiscus producing ciguatera toxins,and Prorocentrum producing okadaic acid and dinophysis toxins,have been concerned as serious human poisonings.We explored the benthic dinoflagellate biodiversity and distribution characteristics of a series of tropical reefs in 20-40-m water depth in wet season in the Zhongsha Islands in South China Sea using morphological,phylogenetic,and cell counting methods.Results show that benthic dinoflagellates in the islands are rich in biodiversity and 15 species from genera Amphidinium,Coolia,Ostreopsis,and Prorocentrum were identifi ed:Amphidinium carterae,A.magnum,A.massartii,A.operculatum,Coolia canariensis,C.malayensis,C.palmyrensis,C.tropicalis,Ostreopsis cf.ovata,Prorocentrum concavum,P.cf.sculptile,P.emarginatum,P.hof fmannianum,P.lima,and P.rhathymum.Among them,A.magnum is reported for the first time in Chinese waters.The abundance of benthic dinoflagellates was relatively low at 88-4345 cells/100 cm^(2) on sediment and 10-91 cells/g on macroalgae.Prorocentrum and Amphidinium were the dominant and subdominant genera,respectively.It is speculated that the low abundance of benthic dinoflagellates is closely related to the scarcity of macroalgae and stronger water motion at the depth>15 m in Zhongsha Islands.This study expanded the study in biodiversity of benthic dinoflagellates in Chinese waters,and revealed the distribution characteristics of harmful benthic microalgae in reef habitats.

Validation of reference genes for gene expression studies in the dinoflagellate Akashiwo sanguinea by quantitative real-time RT-PCR

The accurate measurement of gene expression via quantitative real-time reverse transcription PCR（q RT-PCR）heavily relies on the choice of valid reference gene（s） for data normalization. Resting cyst is the dormant stage in the life cycle of dinoflagellate, which plays crucial roles in HAB-forming dinoflagellate ecology. However, only limited investigations have been conducted on the reference gene selection in dinoflagellates. Gap remained in our knowledge about appropriate HKGs for normalizing gene expression in different life stages, which laid obstacles for the application of q RT-PCR to the HAB-forming group. In this study, six candidate reference genes,18 S ribosomal RNA（18S）, glyceraldehyde-3-phosphate dehydrogenase（GAPDH）, α-tubulin（TUA）, β-tubulin（TUB）, actin（ACT） and cytochrome oxidase subunit 1（COX1）, were evaluated for their expression stability with q RT-PCR and three statistical algorithms（Ge Norm, Norm Finder, and Best Keeper） for the cosmopolitan, harmful algal bloom-forming dinoflagellate Akashiwo sanguinea. Expression patterns were observed across 18 biological samples, including cells at resting stages（resting cysts）, different growth stages, in darkness, exposed to abscisic acid（ABA） and exposed to temperature stress. The results indicated that TUA, 18 S and GAPDH were relatively stable across all tested scenarios. While the best-recommended reference genes differed across experimental groups, the pairs of ACT and TUA, 18 S and GAPDH were the most reliable for cells at different growth stages and darkness treatment. The combination of TUA and TUB was the best choice for normalization in resting cysts and in ABA treatment, respectively. The pair of ACT and COX1 was suitable for temperature treatments. This study was the first to investigate the stable internal reference genes in dinoflagellates at different stages of life cycle,particularly in resting cysts. Our results provided useful information for selection of reference genes in dinoflagellates regarding quantification of gene expression at different experimental scenarios, which will facilitate more accurate and widespread use of q RT-PCR in gene analysis of dinoflagellates and help to design primers targeting orthologous genes in other algal species.

The impact of biotic and abiotic factors on the distribution of surface sediment dinoflagellate cyst assemblages on the Nanji Island in the East China Sea

The dinoflagellate cyst assemblages on the Nanji Island in the East China Sea, are documented at the first time to construct a quantitative overview of the cyst bank from 2014 to 2015. Thirty-four morphotypes from six groups are identified and quantified at eight sampling sites around the island, including a high proportion of potentially toxigenic species(14%). Autotrophic dinocysts constitute 74% of the total cyst counts, which is relatively low(two to thirty-three per millilitre sediment) compared with previous studies in adjacent areas. Scrippsiella trochoidea and Protoperidinium avellana are the most abundant autotrophic and heterotrophic species, respectively. A multivariate analysis is performed to assess associations between dinocysts and abiotic or biotic variables.Differentiation among seasons is evident in the detrended correspondence analysis(DCA) ordination plot, while a spatial pattern is not clearly revealed despite heterogeneity of the hydrodynamic conditions between sampling sites. Soluble reactive phosphate, the ratio of nitrogen to phosphorus concentrations and Karenia mikimotoi bloom are the three factors significantly(P<0.05) related to surface sediment cyst assemblage defined by the canonical correspondence analysis(CCA), highlighting the importance of nutrient regime to a dinocyst distribution in this area. Although attempts to address the origin of HAB events in recent years using seed banks have failed, knowledge can be valuable for further investigation of dinocyst dynamics and potential toxin threats on the Nanji Island.

Acute Toxicity of Formalin to the Red-tide Dinoflagellate Prorocentrum minimum Schiller(Protozoa,Mastigophora)

The acute toxic levels of formalin to a marine red-tide dinoflagellate Prorocentrum minimum Schiller (Mastigophora, Dinoflagellida) were determined by linear regression analysis. The data obtained revealed that there is a close regression relationship between logarithmic concentrations of formalin and mortality probit scales of the organism when exposure times are 1, 2, 6, 12, 24h. The median lethal concentrations (LC50 values) obtained from correlation analysis for these exposure times (with 95% confidence intervals), were 11.83, 6.76, 4.37, 4.27 and 3.98mgL-1, respectively. A toxicity curve was obtained to connect the exposure time and LC50 value from the correlation between them. The results indicated that P. minimum could be killed or fixed by formalin in the concentration range of 4-12mgL-1 within 1-24h.

Assessment of the consecutive harmful dinoflagellate blooms during 2015 in the Izmit Bay(the Marmara Sea)

A series of red tides were observed during 2015 in the Izmit Bay（the Marmara Sea） which is located in the most industrialized and populated region of Turkey. Six samplings were carried out in this area following the red tides.Nitrite-N, nitrate-N, ammonia, silica and orthophosphate concentrations were analyzed spectrophotometrically.Physicochemical conditions were measured by CTD probe. Plankton quantification was performed using counting chambers under microscopes. Prorocentrum micans was the most abundant species, except on May 14,2015, when Noctiluca scintillans was dominant. The abundance of P. micans reached average 18×10~6 ind./L on May 3, 2015 in the Karamürsel station, simultaneously with elevated levels of NH_3 and o-PO_4^（3–）. The sample was also abundant in dead amphipods（（72±12） ind./L） that had been covered by mucilage aggregates produced by P.micans. The highest biomass（calculated by carbon） was recorded as（268±26.0） mg/L on May 14 in the Hereke station. Beside the anthropogenic wastewater discharges, unknown sources and resuspensions caused increases in nutrient levels. After long term northeaster gusts（35 km/h for 5 d） an upwelling occurred on November 6, 2015 after wind-induced sediment resuspension. Although nutrient discharges remarkably decreased over 30 years through established wastewater treatment plants, harmful phytoplankton blooms still occur. Comparing the present results with other studies in nearby Mediterranean seas reveals that the most intense harmful dinoflagellate bloom in recent years occurred in the Izmit Bay. Therefore, additional protection measures necessary for a cleaner Izmit Bay. These incidents also demonstrate that contaminants, accumulated in sediment,may have long-lasting effects on enclosed marine ecosystems.

Reviving and characterizing three species of dinoflagellate cysts dormant for about 70 years in the East China Sea:Biecheleria brevisulcata,Biecheleriopsis adriatica,and Scrippsiella donghaienis

Many marine dinoflagellates can form resting cysts as a part of their life cycle,and the cysts could be buried in sediment and remained viable for as long as over 150 years.However,only a very limited number of cyst species have been revived from long-buried sediments and investigated in regard to a possible shift in the intra-specific genetic structure of a species detected from the historical record at a particular location.Here,we report a successful germination of three species of resting cysts that were sampled from the depth dated back to 1941±18 AD from a 44-cm sediment core from the East China Sea.Seven isolates were established from germination of single cyst isolation or multi-cyst germinations.LSU rRNA gene or ITS sequences of these strains were obtained,then they were identified to be Biecheleria brevisulcata(five strains),Biecheleriopsis adriatica(one strain),and Scrippsiella donghaienis(one strain)in terms of morphology and rRNA gene sequence.Biecheleria brevisulcata strain 1,Bps.adriatica strain 21,and S.donghaienis strain 23 were examined in detail with light microscope(LM)and scanning electron microscope(SEM),and analyzed with high performance liquid chromatography(HPLC)for their pigment compositions,and genetic diversity.We also confirmed the presence of a resting cyst of Bps.adriatica in the field for the first time.The LSU rRNA gene-based genetic distances of Bps.adriatica from that obtained from water sample,single-cell PCR sequencing for the cysts isolated from the surface sediment of the same sea area and that reported from other regions during the recent years,and ITS-based genetic distances of S.donghaienis from that obtained from cysts isolated from the surface sediment of the same location and that reported from other regions during the recent years indicated that the intra-specific genetic structure of each species in the sampling area may have shifted during the last 70 years.Our work confirms that B.brevisulcata,Bps.adriatica,and S.donghaienis,all described as new species around 2010,have inhabited the East China Sea for about 70 years.The present work reports for the first time the revival of dinoflagellate resting cysts long-buried in the coastal sediments of China,which facilitates further study on the historical occurrences of other harmful dinoflagellates and their relevance to the regional climate and environmental changes in China.