Raman spectroscopic insights into the chemical gradients within the wound plug of the green alga Caulerpa taxifolia.

The invasive unicellular green macroalga Caulerpa taxifolia has spread dramatically in the Mediterranean Sea over the last decades. Its success is based on rapid plug formation after wounding, to prevent the loss of cell content. This quick and efficient process involves the rapid transformation of the secondary metabolite caulerpenyne to the reactive 1,4-dialdehyde oxytoxin 2, which acts as a protein crosslinker. The main metabolites of the wound plug were identified as proteins, caulerpenyne derivatives, and sulfated polysaccharides. Because of a methodological deficit, however, the detailed distribution of the compounds within the wound plug of C. taxifolia was unknown. This study demonstrates the suitability of FT-Raman spectroscopy for the noninvasive in vivo determination of caulerpenyne and its derivatives, as well as ß-carotene, from signals with special spectral features within the wound plug and the adjacent intact alga tissue, with a resolution of 100 µm. FT-Raman spectra allowed four different zones with distinct chemical compositions around the region of wounds to be characterized. Gradients of the investigated metabolites within the wound plug and the alga could be determined. Moreover, various caulerpenyne derivatives could be identified spectroscopically, and this has led to a mechanistic proposal for the internal and the external wound plug formation.

Toward in vivo chemical imaging of epicuticular waxes.

Epicuticular waxes, which are found on the outer surface of plant cuticles, are difficult to study in vivo. To monitor the growth, development, and structural alterations of epicuticular wax layers, coherent anti-Stokes Raman scattering (CARS) might be used. CARS, as a Raman-based technique, not only provides structural insight but also chemical information by imaging the spatial distribution of Raman-active vibrations. Here, we present a comparative study using CARS and scanning electron microscopy to characterize the structure of epicuticular waxes. The ability of CARS to provide detailed structural information on the biologically important wax layer was detailed on the examples of cherry laurel (Prunus laurocerasus), hoya (Hoya carnosa), and ceriman/Swiss cheese plant (Monstera sp. aff. deliciosa). We anticipate that the work presented will open a doorway for online monitoring of formation and alterations of epicuticular wax layers.