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Expanding the Symbiodinium (Dinophyceae, Suessiales) Toolkit Through Protoplast Technology

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dc.contributor Ctr Marine Bioinnovat
dc.contributor University of Technology Sydney
dc.contributor Univ New South Wales
dc.contributor Univ Technol Sydney
dc.contributor University of New South Wales Sydney
dc.contributor Univ Melbourne
dc.contributor Australian Institute of Marine Science
dc.contributor Sch Biol Earth & Environm Sci
dc.contributor Australian Inst Marine Sci
dc.contributor Sch BioSci
dc.contributor Sydney Inst Marine Sci
dc.contributor Climate Change Cluster
dc.contributor University of Melbourne
dc.contributor.author van Oppen, Madeleine J. H.
dc.contributor.author Nitschke, Matthew R.
dc.contributor.author Levin, Rachel A.
dc.contributor.author Suggett, David J.
dc.contributor.author Steinberg, Peter D.
dc.date.accessioned 2017-11-05T18:59:12Z
dc.date.accessioned 2017-11-15T03:09:39Z
dc.date.available 2017-11-05T18:59:12Z
dc.date.available 2017-11-15T03:09:39Z
dc.date.issued 2017-09-01
dc.identifier.citation Levin RA, Suggett DJ, Nitschke MR, van Oppen MJH, Steinberg PD (2017) Expanding the Symbiodinium (Dinophyceae, Suessiales) toolkit through protoplast technology. Journal of Eukaryotic Microbiology 64(5): 588-597
dc.identifier.issn 1066-5234
dc.identifier.uri http://epubs.aims.gov.au/11068/11662
dc.description.abstract Dinoflagellates within the genus Symbiodinium are photosymbionts of many tropical reef invertebrates, including corals, making them central to the health of coral reefs. Symbiodinium have therefore gained significant research attention, though studies have been constrained by technical limitations. In particular, the generation of viable cells with their cell walls removed (termed protoplasts) has enabled a wide range of experimental techniques for bacteria, fungi, plants, and algae such as ultrastructure studies, virus infection studies, patch clamping, genetic transformation, and protoplast fusion. However, previous studies have struggled to remove the cell walls from armored dinoflagellates, potentially due to the internal placement of their cell walls. Here, we produce the first Symbiodinium protoplasts from three genetically and physiologically distinct strains via incubation with cellulase and osmotic agents. Digestion of the cell walls was verified by a lack of Calcofluor White fluorescence signal and by cell swelling in hypotonic culture medium. Fused protoplasts were also observed, motivating future investigation into intra-and inter-specific somatic hybridization of Symbiodinium. Following digestion and transfer to regeneration medium, protoplasts remained photosynthetically active, regrew cell walls, regained motility, and entered exponential growth. Generation of Symbiodinium protoplasts opens exciting, new avenues for researching these crucial symbiotic dinoflagellates, including genetic modification.
dc.description.sponsorship The Centre for Marine Bio-Innovation at The University of New South Wales contributed financial support for this study. The Biomedical Imaging Facility at The University of New South Wales covered expenses for the use of the Olympus FV1000 confocal microscope. Iveta Slapetova provided assistance with confocal imaging. Dave Hughes provided assistance with FRRf measurements. Caitlin Lawson provided assistance with Symbiodinium culturing.
dc.language English
dc.subject protoplast fusion
dc.subject STRESS
dc.subject FUSION
dc.subject somatic hybridization
dc.subject GENETIC-TRANSFORMATION
dc.subject SYMBIOTIC ALGAE
dc.subject SOMATIC HYBRID PLANTS
dc.subject dinoflagellate
dc.subject cellulase
dc.subject Cell wall
dc.subject genetic modification
dc.subject THERMAL TOLERANCES
dc.subject cellulose
dc.subject CELL-WALL
dc.subject CLIMATE-CHANGE
dc.subject protoplast generation
dc.subject GENUS SYMBIODINIUM
dc.subject CORAL-REEFS
dc.subject Microbiology
dc.subject zooxanthellae
dc.title Expanding the Symbiodinium (Dinophyceae, Suessiales) Toolkit Through Protoplast Technology
dc.type journal article
dc.identifier.doi 10.1111/jeu.12393
dc.identifier.wos WOS:000410612000004


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