Raman Spectral Analysis in the CHx‐Stretching Region as a Guiding Beacon for Non‐Targeted, Disruption‐Free Monitoring of Germination and Biofilm Formation in the Green Seaweed Ulva

Raman spectroscopy was used to study the complex interactions and morphogenesis of the green seaweed Ulva (Chlorophyta) and its associated bacteria under controlled conditions in a reductionist model system. Integrating multiple imaging techniques contributes to a more comprehensive understanding of these biological processes. Therefore, Raman spectroscopy was introduced as a non‐invasive, label‐free tool for examining chemical information of the tripartite community Ulva mutabilis‐Roseovarius sp.‐Maribacter sp. The study explored cell differentiation, cell wall protrusion, and bacterial‐macroalgae interactions of intact algal thalli. Using Raman spectroscopy, the analysis of the CHx‐stretching wavenumber region distinguished spatial regions in Ulva germination and cellular malformations under axenic conditions and upon inoculation with a specific bacterium in bipartite communities. The spectral information was used to guide in‐depth analyses within the fingerprint region and to identify substance classes such as proteins, lipids, and polysaccharides, including evidence for ulvan found in cell wall protrusions. Raman microspectroscopy was utilized to monitor the germination and cell differentiation of the green seaweed Ulva (Chlorophyta) and its accompanying bacteria in the CHx‐stretching wavenumber region. It offered useful information for further in‐depth fingerprint analysis. Cell differentiation, cellular abnormalities, and biofilm formation were all tracked, and distinct chemical substance classes were found.