Abstract
Filamentous nitrogen fixing cyanobacteria are key players in global nutrient cycling, but the relationship between CO2- and N2-fixation and intercellular exchange of these elements remains poorly understood in many genera. Using high-resolution nanometer-scale secondary ion mass spectrometry (NanoSIMS) in conjunction with enriched H13CO3− and 15N2 incubations of Anabaena oscillarioides, we imaged the cellular distributions of C, N and P and 13C and 15N enrichments at multiple time points during a diurnal cycle as proxies for C and N assimilation. The temporal and spatial distributions of the newly fixed C and N were highly heterogeneous at both the intra- and inter-cellular scale, and indicative of regions performing active assimilation and biosynthesis. Subcellular components such as the neck region of heterocycts, cell division septae and putative cyanophycin granules were clearly identifiable by their elemental composition. Newly fixed nitrogen was rapidly exported from heterocysts and was evenly allocated among vegetative cells, with the exception of the most remote vegetative cells between heterocysts, which were N limited based on lower 15N enrichment. Preexisting functional heterocysts had the lowest levels of 13C and 15N enrichment, while heterocysts that were inferred to have differentiated during the experiment had higher levels of enrichment. This innovative approach, combining stable isotope labeling and NanoSIMS elemental and isotopic imaging, allows characterization of cellular development (division, heterocyst differentiation), changes in individual cell composition and cellular roles in metabolite exchange.
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Acknowledgements
We thank Christina Ramon (LLNL) for invaluable assistance in preparing samples for NanoSIMS analysis, Troy Gunderson (USC) for his assistance with the isotope enrichment component of this study and for the IRMS analysis and Larry Nittler (LLNL) for software development. Work was funded in part by the US Department of Energy Office of Biological and Environmental Research Genomics: GTL research program (RP, PKW, JP, SJF, IDH and KHN). Work was performed at LLNL under the auspices of the US Department of Energy under Contract W-7405-Eng-4. DGC also thanks NSF Division of Ocean Sciences for research support.
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Popa, R., Weber, P., Pett-Ridge, J. et al. Carbon and nitrogen fixation and metabolite exchange in and between individual cells of Anabaena oscillarioides. ISME J 1, 354–360 (2007). https://doi.org/10.1038/ismej.2007.44
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DOI: https://doi.org/10.1038/ismej.2007.44
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