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  • 1
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    Molecular characterization and morphology of Cochlodinium strangulatum, the type species of Cochlodinium, and Margalefidinium gen. nov. for C. polykrikoides and allied species (Gymnodiniales, Dinophyceae) (2017)
    Details
    Publication Date: 2022-05-25
    Description: © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Harmful Algae 63 (2017): 32-44, doi:10.1016/j.hal.2017.01.008.
    Description: Photosynthetic species of the dinoflagellate genus Cochlodinium such as C. polykrikoides, one of the most harmful bloom-forming dinoflagellates, have been extensively investigated. Little is known about the heterotrophic forms of Cochlodinium, such as its type species, Cochlodinium strangulatum. This is an uncommon, large (~200 μm long), solitary, and phagotrophic species, with numerous refractile bodies, a central nucleus enclosed in a distinct perinuclear capsule, and a cell surface with fine longitudinal striae and a circular apical groove. The morphology of C. polykrikoides and allied species is different from the generic type. It is a bloom-forming species with single, two or four-celled chains, small cell size (25–40 μm long) with elongated chloroplasts arranged longitudinally and in parallel, anterior nucleus, eye-spot in the anterior dorsal side, and a cell surface smooth with U-shaped apical groove. Phylogenetic analysis based on LSU rDNA sequences revealed that C. strangulatum and C. polykrikoides/C. fulvescens formed two distally related, independent lineages. Based on morphological and phylogenetic analyses, the diagnosis of Cochlodinium is emended and C. miniatum is proposed as synonym of C. strangulatum. The new genus Margalefidinium gen. nov., and new combinations for C. catenatum, C. citron, C. flavum, C. fulvescens and C. polykrikoides are proposed.
    Description: F.G. was supported by the Brazilian Conselho Nacional de Desenvolvimento Científico e Tecnológico [grant number BJT 370646/2013–14]. Support for M.L.R. and D.M.A. was provided through the Woods Hole Center for Oceans and Human Health, National Science Foundation [grant number OCE–1314642] and National Institute of Environmental Health Sciences [grant number 1–P01–ES021923–01].
    Keywords: HABs ; Harmful algal blooms ; Molecular phylogenetics ; Red tide ; Toxic Dinoflagellata ; Unarmoured dinoflagellate
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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  • 2
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    Effects of an allelochemical in Phaeodactylum tricornutum filtrate on Heterosigma akashiwo : morphological, physiological and growth effects (2017)
    Details
    Publication Date: 2022-05-25
    Description: © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Chemosphere 186 (2017): 527-534, doi:10.1016/j.chemosphere.2017.08.024.
    Description: The effects of an allelochemical extracted from the culture filtrate of diatom Phaeodactylum tricornutum on the raphidophyte Heterosigma akashiwo were investigated using a series of morphological, physiological and biochemical characters. Growth experiments showed that H. akashiwo was significantly inhibited immediately after exposure to the allelochemical, with many cells rapidly dying and lysing based on microscopic observation. The effects of the allelochemical on the surviving cells were explored using Scanning Electron Microscopy (SEM) and Flow cytometry (FCM), the latter by examination of a suite of physiological parameters (membrane integrity, esterase activity, chlorophyll-a content, membrane potential). The results demonstrate that the membrane of H. akashiwo was attacked by the allelochemical directly, causing cell membrane breakage and loss of integrity. Esterase activity was the most sensitive indicator of the impacts of the allelochemical. Membrane potential and chlorophyll-a content both showed significant decreases following exposure of the Heterosigma cells to high concentrations of the allelochemical for 5 and 6 days. Both were affected, but the membrane potential response was more gradual compared to other effects. The cell size of H. akashiwo did not change compared with the control group. The surviving cells were able to continue to grow and in a few days, re-establish a successful culture, even in the presence of residual allelochemical, suggesting either development of cellular resistance, or the degradation of the chemical.
    Description: The authors wish to thank the National Programme on Global Change and Air-Sea Interaction (Grant No. GASI-03-01-02-01); the National Key Research and Development Program [Grant No. 2016YFC1402101]; the assessment of nanomaterials on biological and ecological effects in the coastal area (Grant No. 201505034).
    Keywords: Allelochemical ; Phaeodactylum tricornutum ; Heterosigma akashiwo ; Flow cytometry ; Physiological characters
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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  • 3
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    Insights into the loss factors of phytoplankton blooms : the role of cell mortality in the decline of two inshore Alexandrium blooms (2017)
    Details
    Publication Date: 2022-05-25
    Description: Author Posting. © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Limnology and Oceanography 62 (2017): 1742–1753, doi:10.1002/lno.10530.
    Description: While considerable effort has been devoted to understanding the factors regulating the development of phytoplankton blooms, the mechanisms leading to bloom decline and termination have received less attention. Grazing and sedimentation have been invoked as the main routes for the loss of phytoplankton biomass, and more recently, viral lysis, parasitism and programmed cell death (PCD) have been recognized as additional removal factors. Despite the importance of bloom declines to phytoplankton dynamics, the incidence and significance of various loss factors in regulating phytoplankton populations have not been widely characterized in natural blooms. To understand mechanisms controlling bloom decline, we studied two independent, inshore blooms of Alexandrium fundyense, paying special attention to cell mortality as a loss pathway. We observed increases in the number of dead cells with PCD features after the peak of both blooms, demonstrating a role for cell mortality in their terminations. In both blooms, sexual cyst formation appears to have been the dominant process leading to bloom termination, as both blooms were dominated by small-sized gamete cells near their peaks. Cell death and parasitism became more significant as sources of cell loss several days after the onset of bloom decline. Our findings show two distinct phases of bloom decline, characterized by sexual fusion as the initial dominant cell removal processes followed by elimination of remaining cells by cell death and parasitism.
    Description: This article is a result of research funded by the National Oceanic and Atmospheric Administration Center for Sponsored Coastal Ocean Research ECOHAB program under award no. NA09NOS4780166 to the University of Texas Marine Science Institute (D.L.E) and the Woods Hole Center for Oceans and Human Health by National Science Foundation (NSF) award no. OCE-1314642 and National Institute of Environmental Health Sciences (NIEHS) award no. 1-P01-ES021923-014 to D.M.A. and M.L B.
    Keywords: Phytoplankton bloom dynamics ; Harmful Algal Blooms (HABs) declines ; Phytoplankton mortality ; Programmed cell death (PCD) ; Life cycle transitions ; Alexandrium fundyense
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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  • 4
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    Assessing the use of artificial substrates to monitor Gambierdiscus populations in the Florida Keys (2017)
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    Publication Date: 2022-05-26
    Description: © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Harmful Algae 68 (2017): 52-66, doi:10.1016/j.hal.2017.07.007.
    Description: Four distinct coastal locations were sampled on a monthly basis near Long Key (Florida Keys, USA) over a 13-month period to study Gambierdiscus population dynamics on different substrates, including four macrophyte species (Dictyota spp., Halimeda spp., Laurencia spp., and Thalassia testudinum) and three artificial substrates (polyvinyl chloride (PVC) tiles, burlap, and fiberglass window screen). Cell densities of Gambierdiscus were generally lower on Dictyota versus Halimeda and Laurencia. Cell densities of Gambierdiscus were significantly correlated among macrophyte hosts in 54% of the comparisons, and between macrophyte hosts and artificial substrates in 72% of the comparisons. Predictive slopes determined from regression analyses between cell densities on artificial substrates and macrophyte hosts indicated that, on an areal basis, fewer cells were present on macrophytes versus artificial substrates (cells cm-2) and that slope variation (error) among the different macrophytes and sites ranged from 5% to 200%, averaging 61% overall. As the data required log-transformation prior to analyses, this level of error translates into two-orders of magnitude in range of estimation of the overall average abundance of Gambierdiscus cells on macrophytes (135 cells g-1 wet weight); 20 to 2690 cells g-1 ww. The lack of consistent correlation among Gambierdiscus cell densities on macrophytes versus artificial substrates, coupled with the high level of error associated with the predictive slope estimations, indicates that extreme caution should be taken when interpreting the data garnered from artificial substrate deployments, and that such deployments should be thoroughly vetted prior to routine use for monitoring purposes.
    Description: Funding for this work was provided by NOAA NOS (Cooperative Agreements NA11NOS478-0060 and NA11NOS4780028).
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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  • 5
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    Differential cellular responses associated with oxidative stress and cell fate decision under nitrate and phosphate limitations in Thalassiosira pseudonana : comparative proteomics (2017)
    Public Library of Science
    Details
    Publication Date: 2022-05-26
    Description: © The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS ONE 12 (2017): e0184849, doi:10.1371/journal.pone.0184849.
    Description: Diatoms are important components of marine ecosystems and contribute greatly to the world's primary production. Despite their important roles in ecosystems, the molecular basis of how diatoms cope with oxidative stress caused by nutrient fluctuations remains largely unknown. Here, an isobaric tags for relative and absolute quantitation (iTRAQ) proteomic method was coupled with a series of physiological and biochemical techniques to explore oxidative stress- and cell fate decision-related cellular and metabolic responses of the diatom Thalassiosira pseudonana to nitrate (N) and inorganic phosphate (P) stresses. A total of 1151 proteins were detected; 122 and 56 were significantly differentially expressed from control under N- and P-limited conditions, respectively. In N-limited cells, responsive proteins were related to reactive oxygen species (ROS) accumulation, oxidative stress responses and cell death, corresponding to a significant decrease in photosynthetic efficiency, marked intracellular ROS accumulation, and caspase-mediated programmed cell death activation. None of these responses were identified in P-limited cells; however, a significant up-regulation of alkaline phosphatase proteins was observed, which could be the major contributor for P-limited cells to cope with ambient P deficiency. These findings demonstrate that fundamentally different metabolic responses and cellular regulations are employed by the diatom in response to different nutrient stresses and to keep the cells viable.
    Description: This study was funded by the National Natural Science Foundation of China (41576138, 41076080, 41576138) to Dr. Jun-Rong Liang; the Woods Hole Center for Oceans and Human Health, National Science Foundation (OCE-1314642) to Dr. DonaldM Anderson; the National Institute of Environmental Health Sciences (1-P01-ES021923- 01) to Dr. DonaldM Anderson; and the ERC Advanced Award Diatomite and ANR project DiaDomOil to Dr. Chris Bowler.
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 6
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    LSU rDNA based RFLP assays for the routine identification of Gambierdiscus species (2017)
    Details
    Publication Date: 2022-05-26
    Description: © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Harmful Algae 66 (2017): 20-28, doi:10.1016/j.hal.2017.04.009.
    Description: Gambierdiscus is a genus of benthic dinoflagellates commonly associated with ciguatera fish poisoning (CFP), which is generally found in tropical or sub-tropical regions around the world. Morphologically similar species within the genus can vary in toxicity; however, species identifications are difficult or sometimes impossible using light microscopy. DNA sequencing of ribosomal RNA genes (rDNA) is thus often used to identify and describe Gambierdiscus species and ribotypes, but the expense and time can be prohibitive for routine culture screening and/or large-scale monitoring programs. This study describes a restriction fragment length polymorphism (RFLP) typing method based on analysis of the large subunit ribosomal RNA gene (rDNA) that can successfully identify at least nine of the described Gambierdiscus species and two Fukuyoa species. The software programs DNAMAN 6.0 and Restriction Enzyme Picker were used to identify a set of restriction enzymes (SpeI, HpyCH4IV, and TaqαI) capable of distinguishing most of the known Gambierdiscus species for which DNA sequences were available. This assay was tested using in silico analysis and cultured isolates, and species identifications of isolates assigned by RFLP typing were confirmed by DNA sequencing. To verify the assay and assess intra-specific heterogeneity in RFLP patterns, identifications of 63 Gambierdiscus isolates comprising ten Gambierdiscus species, one ribotype, and two Fukuyoa species were confirmed using RFLP typing, and this method was subsequently employed in the routine identification of isolates collected from the Caribbean Sea. The RFLP assay presented here reduces the time and cost associated with morphological identification via scanning electron microscopy and/or DNA sequencing, and provides a phylogenetically sensitive method for routine Gambierdiscus species assignment.
    Description: Funding for this study was provided by the U.S. National Oceanic and Atmospheric Administration ECOHAB program (CiguaHAB; Cooperative Agreement NA11NOS4780060, NA11NOS4780028), the China Scholarship Council and Natural Science Foundation of China (No. 41606137, 41606136), and the Guangxi Natural Science Foundation (2015GXNSFCA139003, 2016GXNSFBA380037).
    Keywords: Gambierdiscus ; Ciguatera ; RFLP ; LSU rDNA
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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