Growth and Nitrogen Uptake Kinetics in Cultured Procentrum donghaiense

We compared growth kinetics of Prorocentrum donghaiense cultures on different nitrogen (N) compounds including nitrate, ammonium urea, glutamic acid, dialanine, and cynate. P. donghaiense exhibited standard Monod-type growth kinetics over a range of nitrogen concentrations for all N. Compounds tested. Cultures grown on glu and urea had the highest maximum growth rates. However, cultures grown on cyanate had lower half saturation constants. Nitrogen uptake kinetics were measured in nitrate-deplete and replete batch cultures of P. donghaiense. In nitrate-deplete batch cultures, P. donghaiense exhibited Michaelis-Menten type uptake kinetics for nitrate, ammonium, urea and algal amino acids; uptake was saturated at or below 50 umol N L-1. In nitrate replete batch cultures, ammonium, urea, and algal amino acid uptake kinetics were similar to those measured in nitrate deplete batch cultures. Together, our results demonstrate that P. donghaiense can grow well on a variety of N sources, and exhibits similar uptake kinetics under both nutrient replete and deplete conditions. This may be an important factor facilitating their growth during bloom initiation and development in N-enriched estuaries where many algae compete for bioavailable N. and nutrient environment changes as a result of algal growth

Effects of Temperature, Irradiance and pCO2 on the Growth and Nitrogen Utilization of Prorocentrum Donghaiense

Environmental factors such as temp erature, irradiance, and nitrogen (N) supply affect the growth of Prorocentrum donghaiense, but the interactive effects of these physical factors and the effects of atmospheric CO2 (pCO2) on growth and N uptake have not been examined. We compared growth kinetics of P. donghaiense grown on 4 different N substrates (nitrate [NO3 -], ammonium [NH4 +], urea, and glutamic acid [glu]) with respect to temperature, irradiance, and pCO2. Temperature (15 to 30°C) had a positive effect on growth (max. growth rates: 0.17 to 0.65 d-1; optimal temperature: 25 to 30°C); maximum specific growth rates (μmax) declined when cultures were grown at 30°C. P. donghaiense grew well on all 4 N sources, under irradiances ranging from 10 to 180 μmol quanta m-2 s-1. μmax (2.0 ± 0.1 d-1) was observed in cultures growing with NH4+ as the sole N source in the highest irradiance treatment (180 μmol quanta m-2 s-1). These rates were significantly higher than those measured in cultures grown on NO3 -, urea, and glu (all ∼1.4 d-1). Half-saturation constants (Ks) ranged from 66.5 ± 4.6 to 99.4 ± 6.7 μmol quanta m-2 s-1 for cultures grown with glu or NH4+, respectively, as the sole source of N. Both growth and N uptake rates were higher in cultures grown under elevated pCO2. Our results suggest that P. donghaiense exhibits flexible adaptation for N utilization under broad environmental conditions (temperature, irradiance, pCO2), which may play an important role in the formation and duration of P. donghaiense blooms

Metagenomic Sequencing Identifies Highly Diverse Assemblages of Dinoflagellate Cysts in Sediments From Ships\u27 Ballast Tanks

Ships\u27 ballast tanks have long been known as vectors for the introduction of organisms. We applied next-generation sequencing to detect dinoflagellates (mainly as cysts) in 32 ballast tank sediments collected during 2001-2003 from ships entering the Great Lakes or Chesapeake Bay and subsequently archived. Seventy-three dinoflagellates were fully identified to species level by this metagenomic approach and single-cell polymerase chain reaction (PCR)-based sequencing, including 19 toxic species, 36 harmful algal bloom (HAB) forming species, 22 previously unreported as producing cysts, and 55 reported from ballast tank sediments for the first time (including 13 freshwater species), plus 545 operational taxonomic units (OTUs) not fully identified due to a lack of reference sequences, indicating tank sediments are repositories of many previously undocumented taxa. Analyses indicated great heterogeneity of species composition among samples from different sources. Light and scanning electron microscopy and single-cell PCR sequencing supported and confirmed results of the metagenomic approach. This study increases the number of fully identified dinoflagellate species from ballast tank sediments to 142 (\u3e 50% increase). From the perspective of ballast water management, the high diversity and spatiotemporal heterogeneity of dinoflagellates in ballast tanks argues for continuing research and stringent adherence to procedures intended to prevent unintended introduction of non-indigenous toxic and HAB-forming species

Morphology, ultrastructure, and molecular phylogeny of Wangodinium sinense gen. et sp. nov. (Gymnodiniales, Dinophyceae) and revisiting of Gymnodinium dorsalisulcum and Gymnodinium impudicum.

The genus Gymnodinium includes many morphologically similar species, but molecular phylogenies show that it is polyphyletic. Eight strains of Gymnodinium impudicum, Gymnodinium dorsalisulcum and a novel Gymnodinium-like species from Chinese and Malaysian waters and the Mediterranean Sea were established. All of these strains were examined with light microscopy, scanning electron microscopy and transmission electron microscopy. SSU, LSU and internal transcribed spacers rDNA sequences were obtained. A new genus, Wangodinium, was erected to incorporate strains with a loop-shaped apical structure complex (ASC) comprising two rows of amphiesmal vesicles, here referred to as a new type of ASC. The chloroplasts of Wangodinium sinense are enveloped by two membranes. Pigment analysis shows that peridinin is the main accessory pigment in W. sinense. Wangodinium differs from other genera mainly in its unique ASC, and additionally differs from Gymnodinium in the absence of nuclear chambers, and from Lepidodinium in the absence of Chl b and nuclear chambers. New morphological information was provided for G. dorsalisulcum and G. impudicum, e.g., a short sulcal intrusion in G. dorsalisulcum; nuclear chambers in G. impudicum and G. dorsalisulcum; and a chloroplast enveloped by two membranes in G. impudicum. Molecular phylogeny was inferred using maximum likelihood and Bayesian inference with independent SSU and LSU rDNA sequences. Our results support the classification of Wangodinium within the Gymnodiniales sensu stricto clade and it is close to Lepidodinium. Our results also support the close relationship among G. dorsalisulcum, G. impudicum, and Barrufeta. Further research is needed to assign these Gymnodinium species to Barrufeta or to erect new genera

Marine Phytoplankton Temperature versus Growth Responses from Polar to Tropical Waters – Outcome of a Scientific Community-Wide Study

"It takes a village to finish (marine) science these days" Paraphrased from Curtis Huttenhower (the Human Microbiome project) The rapidity and complexity of climate change and its potential effects on ocean biota are challenging how ocean scientists conduct research. One way in which we can begin to better tackle these challenges is to conduct community-wide scientific studies. This study provides physiological datasets fundamental to understanding functional responses of phytoplankton growth rates to temperature. While physiological experiments are not new, our experiments were conducted in many laboratories using agreed upon protocols and 25 strains of eukaryotic and prokaryotic phytoplankton isolated across a wide range of marine environments from polar to tropical, and from nearshore waters to the open ocean. This community-wide approach provides both comprehensive and internally consistent datasets produced over considerably shorter time scales than conventional individual and often uncoordinated lab efforts. Such datasets can be used to parameterise global ocean model projections of environmental change and to provide initial insights into the magnitude of regional biogeographic change in ocean biota in the coming decades. Here, we compare our datasets with a compilation of literature data on phytoplankton growth responses to temperature. A comparison with prior published data suggests that the optimal temperatures of individual species and, to a lesser degree, thermal niches were similar across studies. However, a comparison of the maximum growth rate across studies revealed significant departures between this and previously collected datasets, which may be due to differences in the cultured isolates, temporal changes in the clonal isolates in cultures, and/or differences in culture conditions. Such methodological differences mean that using particular trait measurements from the prior literature might introduce unknown errors and bias into modelling projections. Using our community-wide approach we can reduce such protocol-driven variability in culture studies, and can begin to address more complex issues such as the effect of multiple environmental drivers on ocean biota.EL and MKT were in part supported by the National Science Foundation (NSF) grants DEB-0845932 and OCE-0928819. TAR and KAW were supported by NSF grant OCE-0727227. UP was supported by NSF grants OCE-0926711 and OCE-1041038. PWB and RS were supported by the New Zealand Royal Society Marsden Fund and the Ministry of Science and Innovation. RMK and KH were in part supported by National Oceanic and Atmospheric Administration (NOAA) Monitoring and Event Response for Harmful Algal Blooms (MERHAB) grant NA04NOS4780239 and NSF grant OCE-0238347. DAH and FX-F were supported by NSF grants OCE-0942379, OCE-0962309, and OCE-117030687. MRM was partially supported by NSF grant OCE-0722395 and a NOAA The Ecology and Oceanography of Harmful Algal Blooms (ECOHAB) grant NA06NO54780246. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Transcriptomic Analyses of Scrippsiella trochoidea Reveals Processes Regulating Encystment and Dormancy in the Life Cycle of a Dinoflagellate, with a Particular Attention to the Role of Abscisic Acid

Due to the vital importance of resting cysts in the biology and ecology of many dinoflagellates, a transcriptomic investigation on Scrippsiella trochoidea was conducted with the aim to reveal the molecular processes and relevant functional genes regulating encystment and dormancy in dinoflagellates. We identified via RNA-seq 3,874 (out of 166,575) differentially expressed genes (DEGs) between resting cysts and vegetative cells; a pause of photosynthesis (confirmed via direct measurement of photosynthetic efficiency); an active catabolism including β-oxidation, glycolysis, glyoxylate pathway, and TCA in resting cysts (tested via measurements of respiration rate); 12 DEGs encoding meiotic recombination proteins and members of MEI2-like family potentially involved in sexual reproduction and encystment; elevated expressions in genes encoding enzymes responding to pathogens (chitin deacetylase) and ROS stress in cysts; and 134 unigenes specifically expressed in cysts. We paid particular attention to genes pertaining to phytohormone signaling and identified 4 key genes regulating abscisic acid (ABA) biosynthesis and catabolism, with further characterization based on their full-length cDNA obtained via RACE-PCR. The qPCR results demonstrated elevated biosynthesis and repressed catabolism of ABA during the courses of encystment and cyst dormancy, which was significantly enhanced by lower temperature (4 ± 1°C) and darkness. Direct measurements of ABA using UHPLC-MS/MS and ELISA in vegetative cells and cysts both fully supported qPCR results. These results collectively suggest a vital role of ABA in regulating encystment and maintenance of dormancy, akin to its function in seed dormancy of higher plants. Our results provided a critical advancement in understanding molecular processes in resting cysts of dinoflagellates

Different Geographic Strains of Dinoflagellate <i>Karlodinium veneficum</i> Host Highly Diverse Fungal Community and Potentially Serve as Possible Niche for Colonization of Fungal Endophytes

In numerous studies, researchers have explored the interactions between fungi and their hosting biota in terrestrial systems, while much less attention has been paid to the counterpart interactions in aquatic, and particularly marine, ecosystems. Despite the growing recognition of the potential functions of fungi in structuring phytoplankton communities, the current insights were mostly derived from phytoplankton hosts, such as diatoms, green microalgae, and cyanobacteria. Dinoflagellates are the second most abundant group of phytoplankton in coastal marine ecosystems, and they are notorious for causing harmful algal blooms (HABs). In this study, we used high-throughput amplicon sequencing to capture global snapshots of specific fungal assemblages associated with laboratory-cultured marine dinoflagellate. We investigated a total of 13 clonal cultures of the dinoflagellate Karlodinium veneficum that were previously isolated from 5 geographic origins and have been maintained in our laboratory from several months to more than 14 years. The total recovered fungal microbiome, which consisted of 349 ASVs (amplicon sequencing variants, sequences clustered at a 100% sequence identity), could be assigned to 4 phyla, 18 classes, 37 orders, 65 families, 97 genera, and 131 species. The fungal consortium displayed high diversity and was dominated by filamentous fungi and ascomycetous and basidiomycetous yeasts. A core set of three genera among all the detected fungi was constitutively present in the K. veneficum strains isolated from geographically distant regions, with the top two most abundant genera, Thyridium and Pseudeurotium, capable of using hydrocarbons as the sole or major source of carbon and energy. In addition, fungal taxa previously documented as endophytes in other hosts were also found in all tested strains of K. veneficum. Because host–endophyte interactions are highly variable and strongly case-dependent, these fungal taxa were not necessarily genuine endosymbionts of K. veneficum; instead, it raised the possibility that dinoflagellates could potentially serve as an alternative ecological niche for the colonization of fungal endophytes. Our findings lay the foundation for further investigations into the potential roles or functions of fungi in the regulation of the growth dynamics and HABs of marine dinoflagellates in the field

Plasticity and Multiplicity of Trophic Modes in the Dinoflagellate Karlodinium and Their Pertinence to Population Maintenance and Bloom Dynamics

As the number of mixotrophic protists has been increasingly documented, &ldquo;mixoplankton&rdquo;, a third category separated from the traditional categorization of plankton into &ldquo;phytoplankton&rdquo; and &ldquo;zooplankton&rdquo;, has become a new paradigm and research hotspot in aquatic plankton ecology. While species of dinoflagellates are a dominant group among all recorded members of mixoplankton, the trophic modes of Karlodinium, a genus constituted of cosmopolitan toxic species, were reviewed due to their representative features as mixoplankton and harmful algal blooms (HABs)-causing dinoflagellates. Among at least 15 reported species in the genus, three have been intensively studied for their trophic modes, and all found to be phagotrophic. Their phagotrophy exhibits multiple characteristics: (1) omnivority, i.e., they can ingest a variety of preys in many forms; (2) flexibility in phagotrophic mechanisms, i.e., they can ingest small preys by direct engulfment and much bigger preys by myzocytosis using a peduncle; (3) cannibalism, i.e., species including at least K. veneficum can ingest the dead cells of their own species. However, for some recently described and barely studied species, their tropical modes still need to be investigated further regarding all of the above-mentioned aspects. Mixotrophy of Karlodinium plays a significant role in the population dynamics and the formation of HABs in many ways, which thus deserves further investigation in the aspects of physiological ecology, environmental triggers (e.g., levels of inorganic nutrients and/or presence of preys), energetics, molecular (genes and gene expression regulations) and biochemical (e.g., relevant enzymes and signal molecules) bases, origins, and evaluation of the advantages of being a phagotroph

Toxic and non-toxic dinoflagellates host distinct bacterial communities in their phycospheres

Abstract Dinoflagellates encompass highly abundant and diverse toxin-producing species among marine phytoplankton. Previous works suggested that some bacterial taxa may affect toxins production in dinoflagellates, however, little is known about impact of toxic dinoflagellates on their co-existing bacterial flora. Here we characterized the bacterial communities associated with 22 clonal cultures of dinoflagellates using DNA metabarcoding method, including 11 confirmed toxic species and 11 species that have not been reported to be toxic. Beta diversity analysis revealed that all the 11 species in toxic group were clustered together and distinctly separated from non-toxic group, strongly suggesting the bacterial community composition was largely influenced by toxic dinoflagellate hosts. The toxic group was found to include higher relative abundance of non-carbohydrate utilizers and xenobiotic-degrading taxa and showed inhibitory effects on algicidal bacteria. These findings suggested that toxic dinoflagellates host bacterial communities distinctive from non-toxic species in multiple ways in their phycospheres