Interrogating marine virus-host interactions and elemental transfer with BONCAT and nanoSIMS-based methods

Area of Science:

• Marine microbiology
• Biogeochemistry
• Viral ecology

Background:

• Marine viruses significantly impact ocean ecosystems, yet their direct activity and role in nutrient cycling remain understudied.
• Understanding virus-host dynamics at the single-virus level is crucial for a comprehensive view of marine microbial processes.

Purpose of the Study:

• To introduce and validate two novel techniques, nanoSIMS and BONCAT, for studying marine virus activity and biogeochemical influence.
• To enable direct, quantitative measurements of viral production and nutrient assimilation at the single-virus level.

Main Methods:

• Stable isotope probing coupled with nanoscale secondary ion mass spectrometry (nanoSIMS) to trace nutrient incorporation into viral particles.
• Fluorescence-based biorthogonal non-canonical amino acid tagging (BONCAT) for direct quantification of viral production and detection of newly synthesized viral proteins.
• Application of these methods to model marine systems (Emiliania huxleyi, Synechococcus sp.) and a pilot field study.

Main Results:

• nanoSIMS successfully resolved carbon and nitrogen enrichment in viral particles, providing quantitative estimates of virus production.
• BONCAT enabled direct quantification of viral production with minimal sample manipulation and detected newly synthesized viral proteins.
• Both techniques proved effective in ecologically relevant model systems and initial field applications.

Conclusions:

• nanoSIMS and BONCAT are powerful, complementary tools for advancing single-virus level research in marine environments.
• These methods fill critical knowledge gaps regarding the biogeochemical impact of marine viruses.
• Direct observation of viral activity will improve our understanding of marine nutrient cycling and microbial dynamics.