Dataset: Data and code from an examination of growth rates of cyanobacteria co-cultured with a heterotrophic bacterium, Alteromonas, under either present-day or predicted future pCO2 conditions

Final no updates expectedDOI: 10.26008/1912/bco-dmo.925841.1Version 1 (2024-04-24)Dataset Type:experimental

Principal Investigator: James Jeffrey Morris (University of Alabama at Birmingham)

Scientist: Elizabeth Entwistle (University of Alabama at Birmingham)

Scientist: Zhiying Lu (University of Alabama at Birmingham)

Student: Matthew Kuhl (University of Alabama at Birmingham)

Technician: Alexander Durrant (University of Alabama at Birmingham)

BCO-DMO Data Manager: Shannon Rauch (Woods Hole Oceanographic Institution)


Program: Science, Engineering and Education for Sustainability NSF-Wide Investment (SEES): Ocean Acidification (formerly CRI-OA) (SEES-OA)

Project: Impacts of Evolution on the Response of Phytoplankton Populations to Rising CO2 (P-ExpEv)

Project: Collaborative Research: Ecology and Evolution of Microbial Interactions in a Changing Ocean (LTPE)


Abstract

The CO2 content of Earth's atmosphere is rapidly increasing due to human consumption of fossil fuels. Models based on short-term culture experiments predict that major changes will occur in marine phytoplankton communities in the future ocean, but these models rarely consider how the evolutionary potential of phytoplankton or interactions within marine microbial communities may influence these changes. Here we experimentally evolved representatives of four phytoplankton functional types (silicif...

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Detailed methods can be found in the manuscript "Marine phytoplankton and heterotrophic bacteria rapidly adapt to future pCO2 conditions in experimental co-cultures". A summary of major methods are provided here.

The phytoplankton used in this study as well as the media in which they were grown are Prochlorococcus MIT9312 (PEv medium), Synechococcus CC9311 (SEv medium), Synechocystis PCC6803 (SEv medium), Thalassiosira oceanica CCMP1005 (FEv medium), and Emiliania huxleyi CCMP371 (FEv medium). All media types were derived from media commonly used to cultivate each organism detailed in "Algal Culturing Techniques" edited by Andersen. Prior to use in experiments, phytoplankton cultures were rendered clonal and axenic, then mixed with pure cultures of the heterotrophic bacterium Alteromonas EZ55, obtained by streaking for isolation on YTSS agar.

Cultures were experimentally evolved for approximately 500 generations at 22 degrees Celisius (°C) under approximately 75 micromoles photons per square meter per second (µmol photons m-2 s-1) in acid-washed conical-bottom glass tubes with airtight caps where the carbonate system was manipulated by the addition of acid or base to achieve present-day (400 parts per million (ppm)) or year 2100 (800 ppm) pCO2 conditions. Phytoplankton growth was measured every 48 hours using a Guava HT1 flow cytometer equipped with a 488 nanometer (nm) laser. When phytoplankton cell densities crossed a cutoff value, cultures were diluted 26-fold into fresh media, representing log₂26 or 4.7 generations per transfer. Samples from each lineage were cryopreserved every 25 generations and again at the end of the experiment.

At the end of the evolution period, we subcultured clonal evolved Alteromonas strains by spread-plating evolved cultures on YTSS agar and selecting single, isolated colonies for growth in YTSS broth. Prochlorococcus was rendered axenic by the addition of 100 micrograms per milliliter (µg mL-1) streptomycin. All growth experiments were initiated by mixing axenic Prochlorococcus with a specific Alteromonas clone (or else remaining axenic) and acclimating the co-culture for 3 transfer cycles (approximately 14 generations) at the target pCO2 concentration. Growth was then monitored by flow cytometry as described above for at least 3 subsequent transfers under constant conditions.


Related Datasets

IsRelatedTo

Dataset: The Long Term Phytoplankton Evolution Experiment: Genomic Analysis
Morris, J. J., Entwistle, E., Lu, Z. (2024) Data and analysis code used to experimentally evolve representatives of four phytoplankton functional types in co-culture with a heterotrophic bacterium under either present-day or predicted future pCO2 conditions. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2024-04-25 doi:10.26008/1912/bco-dmo.925872.1

Related Publications

Results

Lu, Z., Entwistle, E., Kuhl, M. D., Durrant, A. R., Filho, M. M. B., Goswami, A., & Morris, J. J. (2024). Marine phytoplankton and heterotrophic bacteria rapidly adapt to future pCO2conditions in experimental co-cultures. https://doi.org/10.1101/2024.02.07.579367
Methods

Hennon, G. M., Morris, J. J., Haley, S. T., Zinser, E. R., Durrant, A. R., Entwistle, E., … Dyhrman, S. T. (2017). The impact of elevated CO2 on Prochlorococcus and microbial interactions with “helper” bacterium Alteromonas. The ISME Journal, 12(2), 520–531. doi:10.1038/ismej.2017.189.
Methods

R. Andersen, Ed., Algal Culturing Techniques, (Academic Press, Burlington, MA, 2005), pp. 596.