However, they may also enhance the secondary colonisation of other bacteria, or the larvae and spores of macrofoulers 9. The epiphytic bacterial community modulates algal growth, morphogenesis, reproduction and chemical defense against other micro- and macrofoulers 1, 2, 8. The epibiosis has both beneficial and detrimental effects on the host. The algae-associated bacterial communities are host-species-specific they differ between algal species 4, 7 and show temporal variation 5. Although the bacterial colonisers are recruited from the surrounding water and vary with season 4, 5, the composition of the macroalgal biofilms differs from that of the seawater 6, 7. Macroalgae host a dense bacterial epibiome forming surface biofilms 2, 3, 4. Particularly bacteria play major structural and functional roles as epibionts and are the primary colonisers 2. Macroalgal surfaces represent a physical interface to the surrounding seawater and are prone to rapid colonisation resulting in fouling and overgrowth 1, 2. This is the first study analysing the seaweed surface microbiome in conjunction with untargeted surface metabolomics and spatial metabolomics approaches. The UPLC-MS, DESI-IMS and NMR analyses failed to confirm the presence of any surface-associated metabolite, except for mannitol, which were previously reported from F. Untargeted spatial metabolomics by DESI-Imaging MS identified the specific localisation and distribution of various primary and secondary metabolites on surface imprints and in algal cross sections. In total, 50 compounds were putatively dereplicated by UPLC-MS/MS, 37 of which were previously reported from both seaweeds and microorganisms.
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An untargeted metabolomics approach using molecular networks, in silico prediction and manual dereplication showed the differential metabolome of the surface and the whole tissue extracts. Fungi of the class Eurotiomycetes were visualised for the first time on an algal surface. The amplicon sequencing, SEM and CARD-FISH imaging studies showed Alphaproteobacteria to predominate the epibiotic bacteria. vesiculosus in conjunction with the composition and spatial distribution of its surface metabolome. This study was designed to analyse the surface epibiome of F. The brown alga Fucus vesiculosus is a keystone marine species, which is subject to heavy surface colonisation.
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