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In this study we used denaturing gradient gel electrophoresis, sequencing analysis, and analytical flow cytometry to monitor the dynamics and genetic richness of Emiliania huxleyi isolates and cooccurring viruses during two mesocosm experiments in a Norwegian fjord in 2000 and 2003. We exploited variations in a gene encoding a protein with calcium-binding motifs (GPA) and in the major capsid protein (MCP) gene to assess allelic and genotypic richness within E. huxleyi and E. huxleyi-specific viruses (EhVs), respectively. To our knowledge, this is the first report that shows the effectiveness of the GPA gene for analysis of natural communities of E. huxleyi. Our results revealed the existence of a genetically rich, yet stable E. huxleyi and EhV community in the fjordic environment. Incredibly, the same virus and host genotypes dominated in separate studies conducted 3 years apart. Both E. huxleyi-dominated blooms contained the same six E. huxleyi alleles. In addition, despite the presence of at least six and four EhV genotypes at the start of the blooms in 2000 and 2003, respectively, the same two virus genotypes dominated the naturally occurring infections during the exponential and termination phases of the blooms in both years.

Despite their importance in shaping the structure and function of marine microbial food webs, little is known about factors regulating marine virus abundance. Previous work demonstrated clearance of laboratory‐cultured Emiliania huxleyi virus by the appendicularian Oikopleura dioica; however, the applicability of this interaction to natural virus assemblages was not investigated. Here, we conducted controlled laboratory experiments using O. dioica and mesocosm water containing natural virus assemblages with high densities of virus, and measured removal of virus by O. dioica using both flow cytometry and molecular methods. Bayesian models based on flow cytometry quantification of virus particles demonstrated efficient removal of viruses (mean 90.3 mL ind−1 d−1), with a clearance efficiency of 42.6% relative to food algae. Molecular detection of virus removal by quantification of viral mcp gene copies revealed a mean clearance rate of 68.1 mL ind−1 d−1. Fecal pellets from these experiments demonstrated that viruses in fecal pellets retain infectivity despite passage through the O. dioica gut. Shotgun metavirome analysis demonstrated O. dioica removal of large virus groups, notably the Phycodnaviridae. The results demonstrate the removal of E. huxleyi virus from natural virus assemblages by O. dioica and the maintenance of viral infectivity when incorporated into fecal pellets, prompting further investigation on the fate of fecal‐packaged viruses and their impact on host dynamics. Furthermore, our results indicate the generality of this interaction for other large algal viruses, raising questions about the implications of this mechanism of marine virus redistribution on the broader marine virus community. [ABSTRACT FROM AUTHOR]

Abstract: Appendicularians are planktonic marine tunicates with elaborate filter‐feeding houses that can efficiently trap particles as small as 0.2 μm. While marine viruses are seldom considered outside their role in disease transmission, we conducted a controlled laboratory experiment to determine if the appendicularian Oikopleura dioica can trap and ingest the Emiliania huxleyi virus (EhV; 160–180 nm diameter). Removal and retention of EhV during 2.5 h and overnight incubations at 15°C were measured using flow cytometry and quantitative polymerase chain reaction specific for the mcp gene of EhV. The fate of retained EhV was tested by quantifying EhV DNA in three biological compartments: house‐trapping, ingestion/digestion, and defecation. Clearance rates for EhV varied from approximately 2 mL ind−1 d−1 to 50 mL ind−1 d−1, with highest rates for 4–5 d‐old animals. EhV particles were cleared by O. dioica at rates similar to those reported for larger food particles, with mean clearance rates in the 2.5 h incubations ranging from approximately 2 mL ind−1 d−1 to 50 mL ind−1 d−1. This demonstrates efficient virus removal by O. dioica and a previously overlooked link between the microbial loop and the classical marine food web. EhV DNA was readily detectable above background levels in O. dioica houses, gut contents, and faecal pellets, suggesting that appendicularian houses and faecal pellets may contribute to the dispersal of viruses. Furthermore, clearance of EhV and presumably other viruses by O. dioica may be a significant sink for viruses and thus an important factor in regulating the population dynamics of viruses and their hosts. [ABSTRACT FROM AUTHOR]

Factors controlling the community composition ofmarine heterotrophic prokaryotes include organic-C, mineral nutrients, predation, and viral lysis. Two mesocosm experiments, performed at an Arctic location and bottom-upmanipulated with organic-C, had very different results in community composition for both prokaryotes and viruses. Previously, we showed how a simple mathematical model could reproduce food web level dynamics observed in these mesocosms, demonstrating strong top-down control through the predator chain from copepods via ciliates and heterotrophic nanoflagellates. Here, we use a steady-state analysis to connect ciliate biomass to bacterial carbon demand. This gives a coupling of top-down and bottom-up factors whereby low initial densities of ciliates are associated withmineral nutrient-limited heterotrophic prokaryotes that do not respond to external supply of labile organic-C. In contrast, high initial densities of ciliates give carbon-limited growth and high responsiveness to organic-C. The differences observed in ciliate abundance, and in prokaryote abundance and community composition in the two experimentswere in accordancewith these predictions. Responsiveness in the viral community followed a pattern similar to that of prokaryotes. Our study provides a unique link between the structure of the predator chain in themicrobial food web and viral abundance and diversity. [ABSTRACT FROM AUTHOR]

Viruses influence the ecology and diversity of phytoplankton in the ocean. Most studies of phytoplankton host-virus interactions have focused on bloom-forming species like Emiliania huxleyi or Phaeocystis spp. The role of viruses infecting phytoplankton that do not form conspicuous blooms have received less attention. Here we explore the dynamics of phytoplankton and algal viruses over several sequential seasons, with a focus on the ubiquitous and diverse phytoplankton division Haptophyta, and their double-stranded DNA viruses, potentially with the capacity to infect the haptophytes. Viral and phytoplankton abundance and diversity showed recurrent seasonal changes, mainly explained by hydrographic conditions. By 454 tag-sequencing we revealed 93 unique haptophyte operational taxonomic units (OTUs), with seasonal changes in abundance. Sixty-one unique viral OTUs, representing Megaviridae and Phycodnaviridae, showed only distant relationship with currently isolated algal viruses. Haptophyte and virus community composition and diversity varied substantially throughout the year, but in an uncoordinated manner. A minority of the viral OTUs were highly abundant at specific time-points, indicating a boom-bust relationship with their host. Most of the viral OTUs were very persistent, which may represent viruses that coexist with their hosts, or able to exploit several host species. [ABSTRACT FROM AUTHOR]

Viruses are ubiquitous components of the marine ecosystem. In the current study we investigated seasonal variations in the viral community in Norwegian coastal waters by pulsed-field gel electrophoresis (PFGE). The results demonstrated that the viral community was diverse, displaying dynamic seasonal variation, and that viral populations of 29 different sizes in the range from 26 to 500 kb were present. Virus populations from 260 to 500 kb and dominating autotrophic pico- and nanoeukaryotes showed similar dynamic variations. Using flow cytometry and real-time PCR, we focused in particular on one host-virus system: Synechococcus spp. and cyanophages. The two groups covaried throughout the year and were found in the highest amounts in fall with concentrations of 7.3 × 104 Synechococcus cells ml-1 and 7.2 × 10³ cyanophage ml-1. By using primers targeting the g20 gene in PCRs on DNA extracted from PFGE bands, we demonstrated that cyanophages were found in a genomic size range of 26 to 380 kb. The genetic richness of the cyanophage community, determined by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified g20 gene fragments, revealed seasonal shifts in the populations, with one community dominating in spring and summer and a different one dominating in fall. Phylogenetic analysis of the sequences originating from PFGE and DGGE bands grouped the sequences into three groups, all with homology to cyanomyoviruses present in cultures. Our results show that the cyanophage community in Norwegian coastal waters is dynamic and genetically diverse and has a surprisingly wide genomic size range. [ABSTRACT FROM AUTHOR]

The isolation and characterization of a virus (designated EhV) that infects the marine coccolithophorid Emiliania huxleyi (Lohmann) Hay & Mohler are described. Three independent clones of EhV were isolated from Norwegian coastal waters in years 1999 and 2000. EhV is a double-stranded DNA-containing virus with a genome size of ∼415 kilo-base pairs. The viral particle is an icosahedron with a diameter of 160–180 nm. The virus particle contains at least nine proteins ranging from 10 to 140 kDa; the major capsid protein weighs ∼54 kDa. EhV has a latent period of 12–14 h and a burst size of 400–1000 (mean, 620) viral particles per cell. A phylogenetic tree based on DNA polymerase amino acid sequences indicates EhV should be assigned to the Phycodnaviridae virus family and that the virus is most closely related to viruses that infect Micromonas pusilla and certain Chlorella species. [ABSTRACT FROM AUTHOR]

A correction to the article " Emerging Interaction Patterns in the Emiliania Huxleyi-EhV System" is presented.

Abstract: The purpose of the present study was to explore the composition and variation of the pico-, nano- and micro-plankton communities in Norwegian coastal waters and Skagerrak, and the co-occurrence of bacteria and viruses. Samples were collected along three cruise transects from Jæren, Lista and Oksøy on the south coast of Norway and into the North Sea and Skagerrak. We also followed a drifting buoy for 55h in Skagerrak in order to observe diel variations. Satellite ocean color images (SeaWiFS) of the chlorophyll a (chl a) distribution compared favorably to in situ measurements in open waters, while closer to the shore remote sensing chl a data was overestimated compared to the in situ data. Using light microscopy, we identified 49 micro- and 15 nanoplankton sized phototrophic forms as well as 40 micro- and 12 nanoplankton sized heterotrophic forms. The only picoeukaryote (0.2–2.0μm) we identified was Resultor micron (Pedinophyceae). Along the transects a significant variation in the distribution and abundance of different plankton forms were observed, with Synechococcus spp and autotrophic picoeukaryotes as the most notable examples. There was no correlation between viruses and chl a, but between viruses and bacteria, and between viruses and some of the phytoplankton groups, especially the picoeukaryotes. Moreover, there was a negative correlation between nutrients and small viruses (Low Fluorescent Viruses) but a positive correlation between nutrients and large viruses (High Fluorescent Viruses). The abundance of autotrophic picoplankton, bacteria and viruses showed a diel variation in surface waters with higher values around noon and late at night and lower values in the evening. Synechococcus spp were found at 20m depth 25–45 nautical miles from shore apparently forming a bloom that stretched out for more than 100 nautical miles from Skagerrak and up the south west coast of Norway. The different methods used for assessing abundance, distribution and diversity of microorganisms yielded complementary information about the plankton community. Flow cytometry enabled us to map the distribution of the smaller phytoplankton forms, bacteria and viruses in more detail than has been possible before but detection and quantification of specific forms (genus or species) still requires taxonomic skills, molecular analysis or both. [Copyright &y& Elsevier]

The isolation and characterization of a virus (designated EhV) that infects the marine coccolithophorid Emiliania huxleyi (Lohmann) Hay & Mohler are described. Three independent clones of EhV were isolated from Norwegian coastal waters in years 1999 and 2000. EhV is a double-stranded DNA-containing virus with a genome size of ∼415 kilo-base pairs. The viral particle is an icosahedron with a diameter of 160–180 nm. The virus particle contains at least nine proteins ranging from 10 to 140 kDa; the major capsid protein weighs ∼54 kDa. EhV has a latent period of 12–14 h and a burst size of 400–1000 (mean, 620) viral particles per cell. A phylogenetic tree based on DNA polymerase amino acid sequences indicates EhV should be assigned to the Phycodnaviridae virus family and that the virus is most closely related to viruses that infect Micromonas pusilla and certain Chlorella species. [ABSTRACT FROM AUTHOR]