Table S1. Primers used in this study. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
are key players in the biogeochemical cycle of iron (Fe) in the ocean, but the capability of different bacterial groups to access this micronutrient is ignored thus far. The aim of our study was to develop a protocol for the combined application of microautoradiography (MICRO) and catalyzed reporter Quizartinib molecular weight deposition–fluorescence in situ hybridization (CARD-FISH) using the radioisotope 55Fe. Among the different washing solutions tested, Ti-citrate-EDTA was the most efficient for the removal of extracellular 55Fe providing sufficiently low background values. We further demonstrate that the washing Angiogenesis inhibitor of cells with Ti-citrate-EDTA and the fixation with paraformaldehyde or formaldehyde do not induce leakage of intracellular 55Fe. Incubating natural bacterial communities collected from contrasting environments, the NW Mediterranean Sea and the Southern Ocean, with 55Fe revealed that 3–29% of bacterial cells were associated with silver grains. Combining microautoradiography with CARD-FISH, we demonstrate that the contribution of different bacterial
groups to total 55Fe-incorporating cells was overall reflected by their relative contribution to abundance. An exception to this pattern was the proportionally higher
contribution of Gammaproteobacteria, SAR86 and Alteromonas. Our study demonstrates the feasibility of MICRO-CARD-FISH using the radiotracer 55Fe and provides the first description of marine bacterial assemblages actively incorporating Fe. second Iron is a rare resource for microorganisms in the ocean. In surface waters, the iron demand of heterotrophic bacteria can be as high as that of phytoplankton, leading to a strong competition among microorganisms (Tortell et al., 1996). Concurrently, heterotrophic bacteria are key players in the remineralization of particulate biogenic and lithogenic iron, thereby contributing to the production of regenerated bioavailable iron (Tortell et al., 1999; Poorvin et al., 2004). Our understanding of the role of heterotrophic bacteria in iron cycling relies mainly on bulk measurements, such as the contribution of bacterial biomass to the biogenic iron stock and bacterial iron uptake rates (Strzepek et al., 2005). By contrast, links between bacterial diversity and biogeochemical functions involving iron are still lacking. Single-cell approaches were proven a powerful tool to study the role of bacterial groups in biogeochemical cycles of the major elements carbon, phosphorus, and sulfur.