Unicellular nitrogen fixing cyanobacteria (UCYN) are abundant members of phytoplankton communities in a wide range of marine environments, including those with rapidly changing nitrogen (N) concentrations. We hypothesized that differences in N availability (N2 vs. combined N) would cause UCYN to shift strategies of intracellular N and C allocation. We used transmission electron microscopy and nanoscale secondary ion mass spectrometry imaging to track assimilation and intracellular allocation of 13C-labeled CO2 and 15N-labeled N2 or NO3 at different periods across a diel cycle in Cyanothece sp. ATCC 51142. We present new ideas on interpreting these imaging data, including the influences of pre-incubation cellular C and N contents and turnover rates of inclusion bodies. Within cultures growing diazotrophically, distinct subpopulations were detected that fixed N2 at night or in the morning. Additional significant within-population heterogeneity was likely caused by differences in the relative amounts of N assimilated into cyanophycin from sources external and internal to the cells. Whether growing on N2 or NO3, cells prioritized cyanophycin synthesis when N assimilation rates were highest. N assimilation in cells growing on NO3 switched from cyanophycin synthesis to protein synthesis, suggesting that once a cyanophycin quota is met, it is bypassed in favor of protein synthesis. Growth on NO3 also revealed that at night, there is a very low level of CO2 assimilation into polysaccharides simultaneous with their catabolism for protein synthesis. This study revealed multiple, detailed mechanisms underlying C and N management in Cyanothece that facilitate its success in dynamic aquatic environments.

Botany Department Federal University of Santa Catarina Campus de Trindade Florianópolis Brazil

Centre for Ecological Research Balaton Limnological Institute Tihany Hungary

Department Experimental Limnology Leibniz Institute of Freshwater Ecology and Inland Fisheries Stechlin Germany

Department of Biology Mount Allison University Sackville NB Canada

Department of Earth Sciences Utrecht University Utrecht Netherlands

Department of Ecology Berlin Institute of Technology Berlin Germany

Department of Microbiology Oregon State University Corvallis OR United States

Global Change Research Institute Czech Academy of Sciences Brno Czechia

Institute of Microbiology Czech Academy of Sciences Centre Algatech Třeboň Czechia

Institute of Parasitology Czech Academy of Sciences Biology Centre České Budějovice Czechia

Laboratoire de Biologie des Organismes et Ecosystèmes Aquatiques FRE 2030 Muséum National d'Histoire Naturelle CNRS IRD Sorbonne Université Université de Caen Normandie Normandie Université Esplanade de la Paix France

Max Planck Institute for Marine Microbiology Bremen Germany

NIOZ Royal Netherlands Institute for Sea Research and Utrecht University Den Burg Netherlands

Sorbonne Université CNRS Laboratoire d'Océanographie de Villefranche Villefranche sur mer France

Sorbonne Université CNRS Laboratoire d'Océanographie Microbienne Banyuls sur mer France

STU UNIVPM Joint Algal Research Center Marine Biology Institute College of Sciences Shantou University Shantou China

Universidade Federal de São Carlos São Carlos Brazil

University of Technology Sydney Climate Change Cluster Faculty of Science Ultimo NSW Australia

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