Production of the cobalamin lower ligand, 5,6-dimethylbenzimidazole, by a diatom under cobalamin-deplete conditions

Abstract

Cobalamin, or vitamin B12, is a micronutrient required by half of all surveyed phytoplankton but produced only by select bacteria and archaea. Cobalamin influences community composition and primary productivity in various regions of the ocean and has been shown to be a critical currency in microbial interactions. Many marine microorganisms can salvage cobinamide and remodel pseudocobalamin, to generate cobalamin using the lower ligand 5,6-dimethylbenzimidazole (DMB), or the ribosylated form, -ribazole. The sources of the cobalamin lower ligand and intermediate remain poorly characterized but are currently attributed to prokaryotes alone. Here, we grew axenic cultures of a recently isolated Thalassiosiraceae diatom over a range of cobalamin concentrations (0, 0.75, 10, 100 pM). Then, using mass spectrometry, we quantified cobalamin and -ribazole in both particulate (cellular) and dissolved (media) phases. We found that this diatom is a facultative cobalamin consumer: it takes up cobalamin when available but can survive in its absence. Strikingly, under low cobalamin availability, we demonstrate that the diatom produces -ribazole. This highlights eukaryotic production as a source of the molecule and suggests that salvaging and remodeling by phytoplankton may be less dependent on prokaryotes than previously imagined. We detected -ribazole in the spent media during stationary phase, highlighting eukaryotic production of the molecule as a public good. We provide evidence that -ribazole is produced by this diatom as a mechanism to cope with low cobalamin availability, suggesting that even amongst organisms that dont absolutely require it, cobalamin use has important consequences for fitness. Importance SectionCobalamin (vitamin B12) is a scarce micronutrient in the ocean that impacts the composition and activity of marine microbial communities. Microbes employ a range of strategies to cope with low cobalamin availability, including remodeling cobalamin intermediates and alternatives into cobalamin when the lower ligand is available. Here we show that a recently isolated Thalassiosiraceae diatom doesnt need cobalamin to survive but will take it up and use it when available. Additionally, the diatom produces the lower ligand (-ribazole) under low cobalamin conditions. The production of -ribazole was previously thought to be restricted to bacteria and archaea, so these results suggest that eukaryotes could play a larger role in cobamide remodeling than previously imagined. This work also demonstrates that even organisms that dont absolutely require cobalamin for survival have adopted metabolic strategies to cope with its absence, emphasizing the importance of this vitamin for phytoplankton metabolism.