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1. Stress responses in an Arctic microalga (Pelagophyceae) following sudden salinity change revealed by gene expression analysis

Author

Nastasia J. Freyria, Thais C. de Oliveira, Mansi Chovatia, Jennifer Johnson, Alan Kuo, Anna Lipzen, Kerrie W. Barry, Igor V. Grigoriev, and Connie Lovejoy

Subjects
Biology (General), QH301-705.5
Abstract

Abstract Marine microbes that have for eons been adapted to stable salinity regimes are confronted with sudden decreases in salinity in the Arctic Ocean. The episodic freshening is increasing due to climate change with melting multi-year sea-ice and glaciers, greater inflows from rivers, and increased precipitation. To investigate algal responses to lowered salinity, we analyzed the responses and acclimatation over 24 h in a non-model Arctic marine alga (pelagophyte CCMP2097) following transfer to realistic lower salinities. Using RNA-seq transcriptomics, here we show rapid differentially expressed genes related to stress oxidative responses, proteins involved in the photosystem and circadian clock, and those affecting lipids and inorganic ions. After 24 h the pelagophyte adjusted to the lower salinity seen in the overexpression of genes associated with freezing resistance, cold adaptation, and salt tolerance. Overall, a suite of ancient widespread pathways is recruited enabling the species to adjust to the stress of rapid salinity change.

Published
2024
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2. Unraveling the transcriptional features and gene expression networks of pathogenic and saprotrophic Ophiostoma species during the infection of Ulmus americana

Author

Thais C. de Oliveira, Nastasia J. Freyria, Jorge Luis Sarmiento-Villamil, Ilga Porth, Philippe Tanguay, and Louis Bernier

Subjects
transcriptome, differential gene expression, Ophiostoma, pathogenicity, host-pathogen interaction, Dutch elm disease, Microbiology, QR1-502
Abstract

ABSTRACTAmerican elm (Ulmus americana), highly prized for its ornamental value, has suffered two successive outbreaks of Dutch elm disease (DED) caused by ascomycete fungi belonging to the genus Ophiostoma. To identify the genes linked to the pathogenicity of different species and lineages of Ophiostoma, we inoculated 2-year-old U. americana saplings with six strains representing three species of DED fungi, and one strain of the saprotroph Ophiostoma quercus. Differential expression analyses were performed following RNA sequencing of fungal transcripts recovered at 3- and 10-days post-infection. Based on a total of 8,640 Ophiostoma genes, we observed a difference in fungal gene expression depending on the strain inoculated and the time of incubation in host tissue. Some genes overexpressed in the more virulent strains of Ophiostoma encode hydrolases that possibly act synergistically. A mutant of Ophiostoma novo-ulmi in which the gene encoding the ogf1 transcription factor had been deleted did not produce transcripts for the gene encoding the hydrophobin cerato-ulmin and was less virulent. Weighted gene correlation network analyses identified several candidate pathogenicity genes distributed among 13 modules of interconnected genes.IMPORTANCEOphiostoma is a genus of cosmopolitan fungi that belongs to the family Ophiostomataceae and includes the pathogens responsible for two devastating pandemics of Dutch elm disease (DED). As the mechanisms of action of DED agents remain unclear, we carried out the first comparative transcriptomic study including representative strains of the three Ophiostoma species causing DED, along with the phylogenetically close saprotrophic species Ophiostoma quercus. Statistical analyses of the fungal transcriptomes recovered at 3 and 10 days following infection of Ulmus americana saplings highlighted several candidate genes associated with virulence and host-pathogen interactions wherein each strain showed a distinct transcriptome. The results of this research underscore the importance of investigating the transcriptional behavior of different fungal taxa to understand their pathogenicity and virulence in relation to the timeline of infection.

Published
2024
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3. Sulfur-cycling chemolithoautotrophic microbial community dominates a cold, anoxic, hypersaline Arctic spring

Author

Elisse Magnuson, Ianina Altshuler, Nastasia J. Freyria, Richard J. Leveille, and Lyle G. Whyte

Subjects
Metagenomics, Metatranscriptomics, Metagenome-assembled genomes, Saline spring, Sulfidic spring, Cryosphere, Microbial ecology, QR100-130
Abstract

Abstract Background Gypsum Hill Spring, located in Nunavut in the Canadian High Arctic, is a rare example of a cold saline spring arising through thick permafrost. It perennially discharges cold (~ 7 °C), hypersaline (7–8% salinity), anoxic (~ 0.04 ppm O2), and highly reducing (~ − 430 mV) brines rich in sulfate (2.2 g.L−1) and sulfide (9.5 ppm), making Gypsum Hill an analog to putative sulfate-rich briny habitats on extraterrestrial bodies such as Mars. Results Genome-resolved metagenomics and metatranscriptomics were utilized to describe an active microbial community containing novel metagenome-assembled genomes and dominated by sulfur-cycling Desulfobacterota and Gammaproteobacteria. Sulfate reduction was dominated by hydrogen-oxidizing chemolithoautotrophic Desulfovibrionaceae sp. and was identified in phyla not typically associated with sulfate reduction in novel lineages of Spirochaetota and Bacteroidota. Highly abundant and active sulfur-reducing Desulfuromusa sp. highly transcribed non-coding RNAs associated with transcriptional regulation, showing potential evidence of putative metabolic flexibility in response to substrate availability. Despite low oxygen availability, sulfide oxidation was primarily attributed to aerobic chemolithoautotrophic Halothiobacillaceae. Low abundance and transcription of photoautotrophs indicated sulfur-based chemolithoautotrophy drives primary productivity even during periods of constant illumination. Conclusions We identified a rare surficial chemolithoautotrophic, sulfur-cycling microbial community active in a unique anoxic, cold, hypersaline Arctic spring. We detected Mars-relevant metabolisms including hydrogenotrophic sulfate reduction, sulfur reduction, and sulfide oxidation, which indicate the potential for microbial life in analogous S-rich brines on past and present Mars. Video Abstract

Published
2023
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4. Salinity tolerance mechanisms of an Arctic Pelagophyte using comparative transcriptomic and gene expression analysis

Author

Nastasia J. Freyria, Alan Kuo, Mansi Chovatia, Jenifer Johnson, Anna Lipzen, Kerrie W. Barry, Igor V. Grigoriev, and Connie Lovejoy

Subjects
Biology (General), QH301-705.5
Abstract

Arctic microalgae show specific adaptation to cold saline arctic conditions with differential expression of several antifreeze proteins, an ice-binding protein and an acyl-esterase involved in cold adaptation.

Published
2022
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5. A decadal perspective on north water microbial eukaryotes as Arctic Ocean sentinels

Author

Nastasia J. Freyria, Nathalie Joli, and Connie Lovejoy

Subjects
Medicine, Science
Abstract

Abstract The North Water region, between Greenland and Ellesmere Island, with high populations of marine birds and mammals, is an Arctic icon. Due to climate related changes, seasonal patterns in water column primary production are changing but the implications for the planktonic microbial eukaryote communities that support the ecosystem are unknown. Here we report microbial community phenology in samples collected over 12 years (2005–2018) from July to October and analysed using high throughput 18S rRNA V4 amplicon sequencing. Community composition was tied to seasonality with summer communities more variable than distinct October communities. In summer, sentinel pan-Arctic species, including a diatom in the Chaetoceros socialis-gelidus complex and the picochlorophyte Micromonas polaris dominated phytoplankton and were summer specialists. In autumn, uncultured undescribed open water dinoflagellates were favored, and their ubiquity suggests they are sentinels of arctic autumn conditions. Despite the input of nutrients into surface waters, autumn chlorophyll concentrations remained low, refuting projected scenarios that longer ice-free seasons are synonymous with high autumn production and a diatom dominated bloom. Overall, the summer sentinel microbial taxa are persisting, and a subset oceanic dinoflagellate should be monitored for possible ecosystem shifts as later autumn ice formation becomes prevalent elsewhere.

Published
2021
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6. Convergent evolution and horizontal gene transfer in Arctic Ocean microalgae

Author

Richard G Dorrell, Alan Kuo, Zoltan Füssy, Elisabeth H Richardson, Asaf Salamov, Nikola Zarevski, Nastasia J Freyria, Federico M Ibarbalz, Jerry Jenkins, Juan Jose Pierella Karlusich, Andrei Stecca Steindorff, Robyn E Edgar, Lori Handley, Kathleen Lail, Anna Lipzen, Vincent Lombard, John McFarlane, Charlotte Nef, Anna MG Novák Vanclová, Yi Peng, Chris Plott, Marianne Potvin, Fabio Rocha Jimenez Vieira, Kerrie Barry, Colomban de Vargas, Bernard Henrissat, Eric Pelletier, Jeremy Schmutz, Patrick Wincker, Joel B Dacks, Chris Bowler, Igor V Grigoriev, Connie Lovejoy, Institut de biologie de l'ENS Paris (IBENS), Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Lawrence Berkeley National Laboratory [Berkeley] (LBNL), Charles University [Prague] (CU), University of Alberta, Université Laval [Québec] (ULaval), HudsonAlpha Genome Sequencing Center, University of Alabama in Huntsville (UAH), Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), King Abdulaziz University, Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), ANR-21-CE02-0014,PanArctica,Caractérisation phylogénétique, écologique et physiologique de le pan-génome des algues Arctiques(2021), and ANR-22-ERCS-0018,ChloroMosaic,Caractérisation moléculaire du réussite environnemental de la chloroplaste secondaire rouge(2022)

Subjects
Ecology, Bacteria, Arctic Regions, Health, Toxicology and Mutagenesis, Oceans and Seas, Gene Transfer, Plant Science, [SDV.BID]Life Sciences [q-bio]/Biodiversity, Biochemistry, Genetics and Molecular Biology (miscellaneous), Horizontal, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Microalgae, Ice Cover, Life Below Water
Abstract

Microbial communities in the world ocean are affected strongly by oceanic circulation, creating characteristic marine biomes. The high connectivity of most of the ocean makes it difficult to disentangle selective retention of colonizing genotypes (with traits suited to biome specific conditions) from evolutionary selection, which would act on founder genotypes over time. The Arctic Ocean is exceptional with limited exchange with other oceans and ice covered since the last ice age. To test whether Arctic microalgal lineages evolved apart from algae in the global ocean, we sequenced four lineages of microalgae isolated from Arctic waters and sea ice. Here we show convergent evolution and highlight geographically limited HGT as an ecological adaptive force in the form of PFAM complements and horizontal acquisition of key adaptive genes. Notably, ice-binding proteins were acquired and horizontally transferred among Arctic strains. A comparison withTaraOceans metagenomes and metatranscriptomes confirmed mostly Arctic distributions of these IBPs. The phylogeny of Arctic-specific genes indicated that these events were independent of bacterial-sourced HGTs in Antarctic Southern Ocean microalgae.

Published
2022

7. Author Reply to Peer Reviews of Convergent evolution and horizontal gene transfer in Arctic Ocean microalgae

Author

Richard G Dorrell, Alan Kuo, Zoltan Fussy, Elisabeth H Richardson, Asaf Salamov, Nikola Zarevski, Nastasia J Freyria, Federico M Ibarbalz, Jerry Jenkins, Juan J Pierella Karlusich, Andrei Stecca Steindorff, Robyn E Edgar, Lori Howell Handley, Kathleen Lail, Anna Lipzen, Vincent Lombard, John McFarlane, Charlotte Nef, Anna MG Novak Vanclova, Yi Peng, Chris Plott, Marianne Potvin, FABIO RJ Vieira, Kerrie Barry, Colomban de Vargas, Bernard Henrissat, Eric Pelletier, Jeremy Schmutz, Patrick Wincker, Joel B. Dacks, Chris Bowler, Igor Vasilij Grigoriev, and Connie Lovejoy

Published
2022

8. A decadal perspective on north water microbial eukaryotes as Arctic Ocean sentinels

Author

Connie Lovejoy, Nastasia J Freyria, and Nathalie Joli

Subjects
0106 biological sciences, Science, Microbial communities, Microbiology, 01 natural sciences, Article, Microbial ecology, 03 medical and health sciences, Water column, Phytoplankton, Ecosystem, 14. Life underwater, 030304 developmental biology, Marine biology, 0303 health sciences, Multidisciplinary, Environmental microbiology, Ecology, biology, 010604 marine biology & hydrobiology, fungi, Chaetoceros, Biodiversity, 15. Life on land, Plankton, biology.organism_classification, Ocean sciences, Diatom, Geography, Biogeography, Arctic, 13. Climate action, Medicine
Abstract

The North Water region, between Greenland and Ellesmere Island, with high populations of marine birds and mammals, is an Arctic icon. Due to climate related changes, seasonal patterns in water column primary production are changing but the implications for the planktonic microbial eukaryote communities that support the ecosystem are unknown. Here we report microbial community phenology in samples collected over 12 years (2005–2018) from July to October and analysed using high throughput 18S rRNA V4 amplicon sequencing. Community composition was tied to seasonality with summer communities more variable than distinct October communities. In summer, sentinel pan-Arctic species, including a diatom in the Chaetoceros socialis-gelidus complex and the picochlorophyte Micromonas polaris dominated phytoplankton and were summer specialists. In autumn, uncultured undescribed open water dinoflagellates were favored, and their ubiquity suggests they are sentinels of arctic autumn conditions. Despite the input of nutrients into surface waters, autumn chlorophyll concentrations remained low, refuting projected scenarios that longer ice-free seasons are synonymous with high autumn production and a diatom dominated bloom. Overall, the summer sentinel microbial taxa are persisting, and a subset oceanic dinoflagellate should be monitored for possible ecosystem shifts as later autumn ice formation becomes prevalent elsewhere.

Published
2021

9. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus.

Author

Emma R Cold, Nastasia J Freyria, Joaquín Martínez Martínez, and José A Fernández Robledo

Subjects
Medicine, Science
Abstract

The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham's F12-5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham's F12-5% FBS- 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham's F12-5% FBS- 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications.

Published
2016
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10. Two versions of short-term phytoplankton ecophysiology and taxonomic assemblages in the Arctic Ocean’s North Water (Canada, Greenland)

Author

Connie Lovejoy, Nathalie Joli, Marcel Babin, Nastasia J Freyria, Thomas Lacour, Sarah-Jeanne Royer, Agencia Estatal de Investigación (España), Takuvik Joint International Laboratory ULAVAL-CNRS, and Université Laval [Québec] (ULaval)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, chemistry.chemical_classification, Ecophysiology, 010504 meteorology & atmospheric sciences, Ecology, 010604 marine biology & hydrobiology, Phytoplankton pigments, Aquatic Science, 01 natural sciences, Term (time), The arctic, Oceanography, chemistry, 13. Climate action, Photoprotection, Xanthophyll, Phytoplankton, [SDE]Environmental Sciences, Environmental science, 14. Life underwater, Ecology, Evolution, Behavior and Systematics, 0105 earth and related environmental sciences
Abstract

16 pages, Photosynthetic performance in open marine waters is determined by how well phytoplankton species are adapted to their immediate environment and available light. Although there is light for 24 h a day during the Arctic summer, little is known about short-term (h) temporal variability of phytoplankton photosynthetic performance in Arctic waters. To address this, we sampled the North Water (76.5°N) every 4 h over 24 h at two stations on the East and West sides that are influenced by different water masses and current conditions. We specifically investigated phytoplankton pigments, the xanthophyll cycle (XC), which is an indication of photoprotective capacity, and photosynthesis–irradiance (PE) response curves, at the surface and 20 m depth. The photophysiological parameters on the two sides differed along with the taxonomic signal derived from accessory pigments. On both sides, surface XC pigments showed high photoprotection capacity with the dinodinoxanthin–diatoxanthin (DD) and the violaxanthin, antheraxanthin and zeaxanthin cycles correlated with incoming radiation. The PE results showed that communities dominated by small flagellates on the western side performed better compared to diatom dominated communities on the eastern side. We conclude that phytoplankton and photosynthetic capacity differed consistent with known hydrography, with implications for a changing Arctic, With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Published
2021

11. Genetic tool development in marine protists: Emerging model organisms for experimental cell biology

Author

Deepak Nanjappa, Iñaki Ruiz-Trillo, Christopher J. Howe, Chris Bowler, Mariusz Nowacki, Anastasios D. Tsaousis, Noelia Lander, Christopher L. Dupont, Shinichiro Maruyama, Fulei Luan, Andrew E. Allen, Anna M. G. Novák Vanclová, Pamela A. Silver, Nastasia J. Freyria, Peter von Dassow, Elisabeth Hehenberger, Konomi Fujimura-Kamada, Julius Lukeš, Roberto Docampo, Deborah L. Robertson, Thomas E. Clemente, Sebastián R. Najle, Kathryn J. Coyne, Cristina Aresté, Brittany N. Sprecher, Lu Wang, Eleanna Kazana, Verónica Freire-Benéitez, Vladimír Hampl, Cecilia Balestreri, Isabel C. Nimmo, Mariana Rius, Yoshihisa Hirakawa, Arnab Pain, Jorge Ibañez, Elin Einarsson, Lev Tsypin, Heriberto Cerutti, Adrian C. Barbrook, G. Jason Smith, Alexandra Z. Worden, Jian Guo, Jean Claude Lozano, Virginia P. Edgcomb, Huan Zhang, Andrea Highfield, Isaac Núñez, Fatma Gomaa, Jan Pyrih, Natalia Ewa Janowicz, Sara J. Bender, Ross F. Waller, Tobias von der Haar, François-Yves Bouget, Matus Valach, Amanda Hopes, Luke M. Noble, Paulo A. Garcia, Nicholas A.T. Irwin, Tamara Matute, Jernej Turnšek, Ian Hu, Sandra Pucciarelli, Fernán Federici, Valérie Vergé, R. Ellen R. Nisbet, Miguel Angel Chiurillo, Mark Moosburner, Monika Abedin Sigg, Aaron P. Turkewitz, Albane Ruaud, Angela Piersanti, Sebastian G. Gornik, Peter R. Girguis, Adam C. Jones, Lawrence A. Klobutcher, Claudio H. Slamovits, Elena Casacuberta, Imen Lassadi, Elizabeth C. Cooney, Kodai Fukuda, Nicole King, Manuel Ares, Jonathan Z. Kaye, Lisa Sudek, Patrick Beardslee, Cristina Miceli, Xiaoxue Wen, Estienne C. Swart, Yuu Ishii, Ambar Kachale, Pia A. Elustondo, Esteban R. Haro-Contreras, Patrick J. Keeling, José A. Fernández Robledo, Glen L. Wheeler, Colin Brownlee, Rowena Stern, Joshua S. Rest, Susana A. Breglia, Senjie Lin, Duncan B. Coles, Jackie L. Collier, Drahomíra Faktorová, David S. Booth, April Woods, Binnypreet Kaur, Thomas Mock, Gertraud Burger, Jun Minagawa, Yutaka Hanawa, Zhu-Hong Li, Rachele Cesaroni, Laboratoire d'Océanographie Microbienne (LOMIC), Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Institut national des sciences de l'Univers (INSU - CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Faktorová, Drahomíra [0000-0001-9623-2233], Nisbet, R. Ellen R. [0000-0003-4487-196X], Allen, Andrew E. [0000-0001-5911-6081], Ares, Manuel, Jr [0000-0002-2552-9168], Bowler, Chris [0000-0003-3835-6187], Burger, Gertraud [0000-0002-8679-8812], Collier, Jackie L. [0000-0001-8774-5715], Cooney, Elizabeth C. [0000-0001-7435-7609], Docampo, Roberto [0000-0003-4229-8784], Dupont, Christopher L. [0000-0002-0896-6542], Edgcomb, Virginia [0000-0001-6805-381X], Freyria, Nastasia J. [0000-0002-4972-9383], Gornik, Sebastian G. [0000-0002-8026-1336], Howe, Christopher J. [0000-0002-6975-8640], Hu, Ian [0000-0002-2287-031X], Ishii, Yuu [0000-0003-1735-9557], Jones, Adam C. [0000-0001-7521-1863], Lin, Senjie [0000-0001-8831-6111], Maruyama, Shinichiro [0000-0002-1128-5916], Minagawa, Jun [0000-0002-3028-3203], Najle, Sebastián R. [0000-0002-0654-9147], Nowacki, Mariusz [0000-0003-4894-2905], Pain, Arnab [0000-0002-1755-2819], Silver, Pamela A. [0000-0002-7856-4071], Jason Smith, G. [0000-0003-1258-4800], Sprecher, Brittany N. [0000-0001-5032-3834], Tsaousis, Anastasios D. [0000-0002-5424-1905], Tsypin, Lev [0000-0002-0642-8468], Turkewitz, Aaron [0000-0003-3531-5806], Turnšek, Jernej [0000-0002-9056-3565], Valach, Matus [0000-0001-8689-0080], von der Haar, Tobias [0000-0002-6031-9254], Mock, Thomas [0000-0001-9604-0362], Worden, Alexandra Z. [0000-0002-9888-9324], Lukeš, Julius [0000-0002-0578-6618], Apollo - University of Cambridge Repository, Nisbet, R Ellen R [0000-0003-4487-196X], Allen, Andrew E [0000-0001-5911-6081], Ares, Manuel [0000-0002-2552-9168], Collier, Jackie L [0000-0001-8774-5715], Cooney, Elizabeth C [0000-0001-7435-7609], Dupont, Christopher L [0000-0002-0896-6542], Freyria, Nastasia J [0000-0002-4972-9383], Gornik, Sebastian G [0000-0002-8026-1336], Howe, Christopher J [0000-0002-6975-8640], Jones, Adam C [0000-0001-7521-1863], Najle, Sebastián R [0000-0002-0654-9147], Silver, Pamela A [0000-0002-7856-4071], Jason Smith, G [0000-0003-1258-4800], Sprecher, Brittany N [0000-0001-5032-3834], Tsaousis, Anastasios D [0000-0002-5424-1905], Worden, Alexandra Z [0000-0002-9888-9324], Gordon and Betty Moore Foundation, Leverhulme Trust, Ministry of Education, Youth and Sports (Czech Republic), and European Commission

Subjects
Resource, Molecular biology, Range (biology), Tree of life (biology), [SDV]Life Sciences [q-bio], Green Fluorescent Proteins, ved/biology.organism_classification_rank.species, Marine Biology, Environment, Diversification (marketing strategy), Biology, Models, Biological, Biochemistry, Genome, 03 medical and health sciences, Transformation, Genetic, 0302 clinical medicine, Species Specificity, Genome editing, 631/1647/767/722, Ecosystem, 14. Life underwater, Model organism, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Dna delivery, 030306 microbiology, ved/biology, Cellular pathways, Eukaryota, Genetic systems, Biodiversity, DNA, Cell Biology, Genetic models, biology.organism_classification, Cell biology, QR, Transformation (genetics), Eukaryote, 030217 neurology & neurosurgery, 631/337, Biotechnology
Abstract

Diverse microbial ecosystems underpin life in the sea. Among these microbes are many unicellular eukaryotes that span the diversity of the eukaryotic tree of life. However, genetic tractability has been limited to a few species, which do not represent eukaryotic diversity or environmentally relevant taxa. Here, we report on the development of genetic tools in a range of protists primarily from marine environments. We present evidence for foreign DNA delivery and expression in 13 species never before transformed and for advancement of tools for eight other species, as well as potential reasons for why transformation of yet another 17 species tested was not achieved. Our resource in genetic manipulation will provide insights into the ancestral eukaryotic lifeforms, general eukaryote cell biology, protein diversification and the evolution of cellular pathways., This collaborative effort was supported by the Gordon and Betty Moore Foundation EMS Program of the Marine Microbiology Initiative (grant nos. GBMF4972 and 4972.01 to F.-Y.B.; GBMF4970 and 4970.01 to M.A. and A.Z.W.; GBMF3788 to A.Z.W.; GBMF 4968 and 4968.01 to H.C.; GBMF4984 to V.H.; GBMF4974 and 4974.01 to C. Brownlee; GBMF4964 to Y. Hirakawa; GBMF4961 to T. Mock; GBMF4958 to P.S.; GBMF4957 to A.T.; GBMF4960 to G.J.S.; GBMF4979 to K.C.; GBMF4982 and 4982.01 to J.L.C.; GBMF4964 to P.J.K.; GBMF4981 to P.v.D.; GBMF5006 to A.E.A.; GBMF4986 to C.M.; GBMF4962 to J.A.F.R.; GBMF4980 and 4980.01 to S.L.; GBMF 4977 and 4977.01 to R.F.W.; GBMF4962.01 to C.H.S.; GBMF4985 to J.M.; GBMF4976 and 4976.01 to C.H.; GBMF4963 and 4963.01 to V.E.; GBMF5007 to C.L.D.; GBMF4983 and 4983.01 to J.L.; GBMF4975 and 4975.01 to A.D.T.; GBMF4973 and 4973.01 to I.R.-T. and GBMF4965 to N.K.), by The Leverhulme Trust (RPG-2017-364) to T. Mock and A. Hopes, and by ERD funds (16_019/0000759) from the Czech Ministry of Education to J.L.

Published
2020

12. Publisher Correction: Genetic tool development in marine protists: emerging model organisms for experimental cell biology

Author

Claudio H. Slamovits, Gertraud Burger, Patrick J. Keeling, Jian Guo, Fatma Gomaa, Jan Pyrih, Jun Minagawa, Christopher L. Dupont, Verónica Freire-Benéitez, Nastasia J. Freyria, Rachele Cesaroni, Elizabeth C. Cooney, Eleanna Kazana, Aaron P. Turkewitz, Peter R. Girguis, Konomi Fujimura-Kamada, Chris Bowler, Anastasios D. Tsaousis, Pamela A. Silver, Lawrence A. Klobutcher, Tamara Matute, Shinichiro Maruyama, Nicole King, Virginia P. Edgcomb, Roberto Docampo, Isaac Núñez, Deborah L. Robertson, Xiaoxue Wen, Zhu-Hong Li, Huan Zhang, Jorge Ibañez, Kathryn J. Coyne, Jonathan Z. Kaye, Christopher J. Howe, Andrew E. Allen, Ross F. Waller, Brittany N. Sprecher, R. Ellen R. Nisbet, Mark Moosburner, Alexandra Z. Worden, Thomas Mock, Angela Piersanti, Sebastian G. Gornik, Paulo A. Garcia, Amanda Hopes, Isabel C. Nimmo, Mariana Rius, Estienne C. Swart, Nicholas A.T. Irwin, Elisabeth Hehenberger, Valérie Vergé, Andrea Highfield, Sandra Pucciarelli, François-Yves Bouget, Yutaka Hanawa, Matus Valach, April Woods, Elin Einarsson, Anna M. G. Novák Vanclová, Lu Wang, Fernán Federici, Monika Abedin Sigg, Adrian C. Barbrook, Yoshihisa Hirakawa, G. Jason Smith, Adam C. Jones, Deepak Nanjappa, Vladimír Hampl, Binnypreet Kaur, Thomas E. Clemente, Jernej Turnšek, Jean Claude Lozano, Sara J. Bender, Ian Hu, Sebastián R. Najle, Imen Lassadi, Glen L. Wheeler, Cecilia Balestreri, Fulei Luan, Lev Tsypin, Heriberto Cerutti, Miguel Angel Chiurillo, Colin Brownlee, Rowena Stern, Julius Lukeš, Manuel Ares, Drahomíra Faktorová, Kodai Fukuda, David S. Booth, Peter von Dassow, Elena Casacuberta, Patrick Beardslee, Cristina Miceli, Cristina Aresté, Natalia Ewa Janowicz, Joshua S. Rest, Susana A. Breglia, Senjie Lin, Luke M. Noble, Albane Ruaud, Duncan B. Coles, Pia A. Elustondo, Jackie L. Collier, Lisa Sudek, Yuu Ishii, José A. Fernández Robledo, Noelia Lander, Tobias von der Haar, Ambar Kachale, Esteban R. Haro-Contreras, Arnab Pain, Iñaki Ruiz-Trillo, and Mariusz Nowacki

Subjects
biology, Molecular biology, media_common.quotation_subject, Garcia, Green Fluorescent Proteins, Drahomira, Eukaryota, Marine Biology, Cell Biology, Art, Biodiversity, DNA, Environment, biology.organism_classification, Genetic models, Biochemistry, Publisher Correction, Models, Biological, Transformation, Genetic, Species Specificity, Humanities, Ecosystem, Biotechnology, media_common
Abstract

Author(s): Faktorova, Drahomira; Nisbet, R Ellen R; Fernandez Robledo, Jose A; Casacuberta, Elena; Sudek, Lisa; Allen, Andrew E; Ares, Manuel; Areste, Cristina; Balestreri, Cecilia; Barbrook, Adrian C; Beardslee, Patrick; Bender, Sara; Booth, David S; Bouget, Francois-Yves; Bowler, Chris; Breglia, Susana A; Brownlee, Colin; Burger, Gertraud; Cerutti, Heriberto; Cesaroni, Rachele; Chiurillo, Miguel A; Clemente, Thomas; Coles, Duncan B; Collier, Jackie L; Cooney, Elizabeth C; Coyne, Kathryn; Docampo, Roberto; Dupont, Christopher L; Edgcomb, Virginia; Einarsson, Elin; Elustondo, Pia A; Federici, Fernan; Freire-Beneitez, Veronica; Freyria, Nastasia J; Fukuda, Kodai; Garcia, Paulo A; Girguis, Peter R; Gomaa, Fatma; Gornik, Sebastian G; Guo, Jian; Hampl, Vladimir; Hanawa, Yutaka; Haro-Contreras, Esteban R; Hehenberger, Elisabeth; Highfield, Andrea; Hirakawa, Yoshihisa; Hopes, Amanda; Howe, Christopher J; Hu, Ian; Ibanez, Jorge; Irwin, Nicholas AT; Ishii, Yuu; Janowicz, Natalia Ewa; Jones, Adam C; Kachale, Ambar; Fujimura-Kamada, Konomi; Kaur, Binnypreet; Kaye, Jonathan Z; Kazana, Eleanna; Keeling, Patrick J; King, Nicole; Klobutcher, Lawrence A; Lander, Noelia; Lassadi, Imen; Li, Zhuhong; Lin, Senjie; Lozano, Jean-Claude; Luan, Fulei; Maruyama, Shinichiro; Matute, Tamara; Miceli, Cristina; Minagawa, Jun; Moosburner, Mark; Najle, Sebastian R; Nanjappa, Deepak; Nimmo, Isabel C; Noble, Luke; Novak Vanclova, Anna MG; Nowacki, Mariusz; Nunez, Isaac; Pain, Arnab; Piersanti, Angela; Pucciarelli, Sandra; Pyrih, Jan; Rest, Joshua S | Abstract: An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Published
2020

13. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus

Author

José A. Fernández Robledo, Joaquín Martínez Martínez, Nastasia J. Freyria, and Emma R. Cold

Subjects
0301 basic medicine, Aquatic Organisms, Life Cycles, lcsh:Medicine, Pathogenesis, Pathology and Laboratory Medicine, Schizogony, Perkinsus marinus, Animal Products, Medicine and Health Sciences, Agar, Parasite hosting, lcsh:Science, Protozoans, Fluids, Multidisciplinary, biology, Physics, Eukaryota, Agriculture, Alveolata, Host-Pathogen Interactions, Physical Sciences, Research Article, States of Matter, food.ingredient, Meat, Parasitic Life Cycles, 030106 microbiology, Research and Analysis Methods, Transfection, Microbiology, Agar plate, 03 medical and health sciences, food, Parasite Groups, Animals, Parasites, 14. Life underwater, Trophozoites, Perkinsus, Molecular Biology Techniques, Molecular Biology, Shellfish, Nutrition, lcsh:R, Organisms, Biology and Life Sciences, Liquids, Subcloning, biology.organism_classification, Parasitic Protozoans, Culture Media, Ham, Diet, 030104 developmental biology, Mollusca, Food, Parasitology, lcsh:Q, Apicomplexa, Cloning, Developmental Biology
Abstract

The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham's F12-5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham's F12-5% FBS- 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham's F12-5% FBS- 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications.

Published
2016

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