Award Abstract # 1829641
Collaborative Research: Inferring Cellular Lysis and Regeneration of Organic Matter by Marine Viruses

NSF Org: OCE
Division Of Ocean Sciences
Recipient: UNIVERSITY OF TENNESSEE
Initial Amendment Date: August 18, 2018
Latest Amendment Date: July 22, 2021
Award Number: 1829641
Award Instrument: Continuing Grant
Program Manager: Daniel J. Thornhill
OCE
 Division Of Ocean Sciences
GEO
 Directorate for Geosciences
Start Date: October 1, 2018
End Date: September 30, 2023 (Estimated)
Total Intended Award Amount: $598,368.00
Total Awarded Amount to Date: $657,818.00
Funds Obligated to Date: FY 2018 = $95,497.00
FY 2019 = $299,856.00

FY 2020 = $203,015.00

FY 2021 = $59,450.00
History of Investigator:
  • Steven Wilhelm (Principal Investigator)
    wilhelm@utk.edu
Recipient Sponsored Research Office: University of Tennessee Knoxville
201 ANDY HOLT TOWER
KNOXVILLE
TN  US  37996-0001
(865)974-3466
Sponsor Congressional District: 02
Primary Place of Performance: The University of Tennessee
1 Circle Park Drive
Knoxville
TN  US  37996-0003
Primary Place of Performance
Congressional District:
02
Unique Entity Identifier (UEI): FN2YCS2YAUW3
Parent UEI: LXG4F9K8YZK5
NSF Program(s): BIOLOGICAL OCEANOGRAPHY
Primary Program Source: 01001819DB NSF RESEARCH & RELATED ACTIVIT
01001920DB NSF RESEARCH & RELATED ACTIVIT

01002021DB NSF RESEARCH & RELATED ACTIVIT

01002122DB NSF RESEARCH & RELATED ACTIVIT
Program Reference Code(s): 097Z, 102Z, 1097, 1315, 1389, 8811, 9117
Program Element Code(s): 165000
Award Agency Code: 4900
Fund Agency Code: 4900
Assistance Listing Number(s): 47.050

ABSTRACT

Viral infections of marine microbes can transform the fate of microbial populations that fuel global ocean biogeochemical cycles. For example, viral infections of microbes lead to the release of carbon and nutrients back into the environment. This regeneration of carbon and nutrients stimulates the activity of other microbes and diverts carbon and nutrients from larger organisms in marine food webs. Because virus-microbe infections are relatively specific, it is critical to identify those pairs of viruses and microbes that may disproportionally contribute to the turnover of carbon and nutrients in the ocean. This project will develop quantitative approaches and tools to quantify which viruses infect which microbes and to use these data to quantify how viral infections of microbes collectively shape nutrient and carbon cycles in the North Atlantic Ocean. The project will analyze virus-microbe interactions in mesocosms at the Bigelow Laboratory for Ocean Sciences in mid-coast Maine and during open ocean expeditions to the Bermuda Atlantic Time-Series Study (BATS) site. An interdisciplinary team will leverage recent advances in molecular biology, computational biology, and mathematical modeling to identify virus-host partners and their impact on the movement of elements through marine systems. This project will support three graduate students, six undergraduate students and one postdoctoral researcher in an interdisciplinary context. Research advances will be translated into reproducible software methods to be disseminated via the community cyberinfrastructure platform iVirus, with additional training materials presented as part of a viral methods and informatics workshop held at The Ohio State University. The translation of discoveries to the public will be furthered by the involvement of journalism undergraduate students at the University of Tennessee-Knoxville.

This project builds upon advances in the molecular toolkit of viromics to develop an integrated approach to characterize lineage-specific rates of infection, lysis, and nutrient release induced by marine viruses in open ocean ecosystems. It will combine theory, in vitro experiments, and in situ sampling to (i) extend a robust inference method for estimating virus-microbe cross-infection networks from time-series data; (ii) establish and characterize in-vitro protocols for inferring cross-infectivity in complex communities using culture-independent methods; (iii) estimate lineage-specific rates of lysis and regeneration of nutrients in marine systems, including applications to coastal and open ocean ecosystems. Project aims focus on quantifying the extent to which virus-induced lysis and regeneration of carbon and nutrients is heterogeneously distributed across microbial populations. To do so, the project will incorporate time series measurements of abundance information (via metagenomes) and activity information (via metatranscriptomes). In so doing, it will advance efforts to understand community-scale interactions rather than those amongst a single virus-host pair. Theoretical methods and in vitro protocols will directly infer lineage-specific infection, lysis, and nutrient release rates in coastal- and open-ocean ecosystems in the North Atlantic Ocean. Results will be used to identify key links that disproportionately influence bulk nutrient release. A novel PCR-based approach will augment and validate the core inference approach. Overall, the project aims to enhance our understanding of how viruses contribute to marine ecosystem function.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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Coy, Samantha R. and Alsante, Alyssa N. and Van Etten, James L. and Wilhelm, Steven W. and Wood, Susanna A "Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host" PLOS ONE , v.14 , 2019 10.1371/journal.pone.0211755 Citation Details
Gilbert, Naomi E. and LeCleir, Gary R. and Pound, Helena L. and Strzepek, Robert F. and Ellwood, Michael J. and Twining, Benjamin S. and Roux, Simon and Boyd, Philip W. and Wilhelm, Steven W. "Giant Virus Infection Signatures Are Modulated by Euphotic Zone Depth Strata and Iron Regimes of the Subantarctic Southern Ocean" mSystems , v.8 , 2023 https://doi.org/10.1128/msystems.01260-22 Citation Details
Gilbert, Naomi E. and LeCleir, Gary R. and Strzepek, Robert F. and Ellwood, Michael J. and Twining, Benjamin S. and Roux, S. and Pennacchio, C. and Boyd, Philip W. and Wilhelm, Steven W. "Bioavailable iron titrations reveal oceanic Synechococcus ecotypes optimized for different iron availabilities" ISME Communications , v.2 , 2022 https://doi.org/10.1038/s43705-022-00132-5 Citation Details
LeCleir, Gary R. and Bassett, Jenna and Wilhelm, Steven W. "Effects of iron concentration and DFB (Desferrioxamine-B) on transcriptional profiles of an ecologically relevant marine bacterium" PLOS ONE , v.18 , 2023 https://doi.org/10.1371/journal.pone.0295257 Citation Details
Muratore, Daniel and Gilbert, Naomi E and LeCleir, Gary R and Wilhelm, Steven W and Weitz, Joshua S "Diel partitioning in microbial phosphorus acquisition in the Sargasso Sea" Proceedings of the National Academy of Sciences , v.122 , 2025 https://doi.org/10.1073/pnas.2410268122 Citation Details
Sofen, Laura E. and Antipova, Olga A. and Ellwood, Michael J. and Gilbert, Naomi E. and LeCleir, Gary R. and Lohan, Maeve C. and Mahaffey, Claire and Mann, Elizabeth L. and Ohnemus, Daniel C. and Wilhelm, Steven W. and Twining, Benjamin S. "Trace metal contents of autotrophic flagellates from contrasting openocean ecosystems" Limnology and Oceanography Letters , v.7 , 2022 https://doi.org/10.1002/lol2.10258 Citation Details

PROJECT OUTCOMES REPORT

Disclaimer

This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.

The project was a collaboration between the University of Tennessee, The Ohio State University and Georgia Institute of Technology/The University of Maryland.  The main goal of the research was to understand how the millions of viruses in every drop of ocean water interact with the hundreds and thousands of bacteria found in the same volume of ocean.  A key goal for the University of Tennessee group was to accomplish this from RNA sequencing data that is presently being broadly collected by researchers around the world. 

Starting with data generated in the spring of 2018 from the Southern Ocean near Tasmania, the team was able to develop strong linkages between the virus communities in the system and their potential hosts.  This was particularly true to a group known as the "giant viruses": these members of the Nucleocytoviricota infect protists that are abundant members of the Southern Ocean community. The observations demonstrated that changes in the availability of the trace element Fe influenced the activity of these viruses.  Increases in Fe availability made some more activty while decreases in the availability of Fe made other viruses more active. The results shine a light on how nutrients can indirectly regulate microbial community form and function.

The team also completed field expeditions to the Atlantic Ocean in 2019 and 2022.  In 2019 a 5 day diel study (i.e., around the clock) sampled several ocean depths every 4 hours.  Team member extracted RNA for sequencing as well as small molecules to characterize microbial community changes with the solar day.  One major observation was the rediscovery of a region of high oxygen just below the thermocline we have refered to as the subsurface oxygen maximum (SOM).  This region receives reduced surface sunlight (~ 1%) while at the same time getting nutrients which are upwelled from below. Our observations suggested that this region is expansive across the Atlantic and that biological activity in it is driven by a subset of the cyanobacteria in the genus Prochlorococcus along with various heterotrophic bacteria.  Surprisingly, it appaears that the more rapid activity in this region is driven by increase virus infection of the Prochlorococcus: increased infection leads to faster carbon cycling (as more cyanobacteria lyse) which leads to more heterotrophic bacterial activity.  The bacteria respire off the carbon and then recycling ammonium, which in turn enhances the growth rate of the Prochlorococcus.  One conclusion from this data is that the SOM is likely strong affected by regional climate, and it could become under threat with predictions of climate change that will alter ocean circulation and stratification. 

A final significant observation in this region was that the assimilation of phosphorous (P) is like partitioned by different members of the microbial community to different times of day.  This observation arose from the diel assessment of microbial community function as observed in the RNA sequencing data.  In short, different members of the community seemed tuned to  best competing for this limiting resource at different times of day.   This suggests very tight regulation of nutrient dynamics across the entire community.  Moving forward the data also suggest that changes in circulation and mixing might alter how cells perceived time of day (e.g., by changing the depth cells mix too in the water column they will get different light cues) and thus the physical changes in the water column brought on by a changing climate might have serious complications for the future.

Along with the above scientific output, this project trained graduate and undergraduate students in lab and field techniques invcluding the state-of-the-art bioinformatics needed to analyse the billions of RNA sequences generated by this study.  Papers have been published and will continue to be published from this work, and our observations have been presented at scientific meetings around the globe. 


Last Modified: 01/29/2024
Modified by: Steven W Wilhelm

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