REMineralisation of organic carbon by marine bActerIoplanktoN (REMAIN) - reducing the known unknown

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The balance between the uptake of CO2 during phytoplankton photosynthesis and the production of CO2 during bacterial, zooplankton and phytoplankton respiration influences how much carbon can be stored in the ocean and hence how much remains in the atmosphere to affect climate. Yet, despite its crucial role, our knowledge of the respiration of component plankton groups such as bacteria, is severely limited because we do not have a method which can differentiate the respiration of one group from that of the rest of the community. We resort to measuring the respiration of a subsample which contains only cells which have passed through a filter. Just as we take in oxygen when we breathe, plankton take in oxygen when they respire and so the standard way to measure respiration is as the decrease in oxygen in the water. Unfortunately the low rates of respiration mean that measurements of oxygen have to be made over many hours and the disruption of the plankton foodweb by the filtration can lead to major errors.The recent development of a much more sensitive method (reduction of the tetrazolium salt INT) which can produce results in minutes and does not involve disruption of the plankton foodweb, is a major step forward and has revealed previously unknown variability. Unexpectedly, the results also suggest that the proportion of respiration attributable to the bacterial size class is consistently low, even in communities where bacteria are the most numerous plankton. This has profound implications for our understanding of the amount of CO2 produced by different plankton groups, and poses two new questions - which size class contributes most to plankton respiration if not the bacterial size class, and what influences the variability in respiration if not the type of plankton present. Marine scientists including ourselves have excitedly started to use the new INT technique, but it has not been thoroughly tested for all plankton communities. In fact, recent data suggest that the method can sometimes underestimate respiration because not all plankton can take up INT and sometimes overestimate respiration because compounds not associated with respiration can affect the INT. Thus while this method could potentially enable a critical improvement in our understanding and thus prediction of CO2 cycling in the ocean, these new intriguing results cannot be confirmed until a comprehensive test of the method has been completed. This is what we will do.We have brought together an international team of experts to undertake an innovative combination of laboratory and field work, taking advantage of a unique sampling opportunity - the Atlantic Meridional Transect - which allows the study of five different plankton ecosystems in the Atlantic Ocean. We will first develop a novel fluorescent method of tracking the uptake of INT into a representative range of plankton to quantify how different cells take it up. Then we will measure the INT reduction of both seawater samples and laboratory cultures to which have been added chemical inhibitors of plankton respiration and increasing concentrations of naturally occurring organic compounds, to determine to what extent these organic compounds lead to an overestimate of respiration. These results will be used to improve and validate the INT method. Finally we will participate in the research cruise to determine plankton respiration in size classes of the plankton community alongside identification of the plankton in these size classes and the concentration of organic compounds. The main deliverable of the project - apportionment of plankton respiration to plankton size classes - is of benefit to marine scientists who aim to predict how a changing climate will affect plankton production of CO2, policy makers interested in how much carbon can be stored in the ocean, and potentially commercial companies interested in the development of the fluorescent probe for medical or water quality applications.