Click & sea: using bioorthogonal click chemistry to visualize seaweed cell walls

Area of Science:

• Marine biology
• Phycology
• Biochemistry

Background:

• Seaweed cell wall research is vital for understanding climate change adaptation.
• Current in situ methods for studying seaweed cell walls have limitations, including a lack of specific antibodies and incomplete characterization of cell wall components.
• Studying seaweed cell wall metabolism in situ is challenging.

Purpose of the Study:

• To develop and present a new method for labeling seaweed cell walls in situ.
• To utilize bioorthogonal click chemistry for visualizing seaweed cell wall components.
• To overcome limitations of existing techniques for studying seaweed cell wall metabolism and composition.

Main Methods:

• Incorporation of chemical reporters (activated sugar analogues of fucose, galactose, and glucose) directly into seaweed tissues (Phycodrys rubens and Ulva spp.).
• Visualization of incorporated reporters using click chemistry to attach a fluorescent probe (AF488).
• Confocal microscopy imaging and image processing (using Fiji software) to analyze specific fluorescence in cell walls.

Main Results:

• Successful incorporation of activated sugars into seaweed cell walls and some organelles was observed.
• Fluorescence was primarily localized to cell walls, indicating specific incorporation into cell wall components like polysaccharides or glycoproteins.
• Image analysis confirmed specific and good incorporation of the fluorochrome after accounting for autofluorescence.

Conclusions:

• Bioorthogonal click chemistry offers a valuable new tool for phycologists studying seaweed cell walls.
• This method can provide deeper insights into seaweed cell wall composition and dynamics.
• The technique is applicable to understanding seaweed responses to various stresses, including those related to climate change.