A laser-scanning confocal microscopy study of carrageenan in red algae from seaweed farms near the Caribbean entrance of the Panama Canal

Kappaphycus alvarezii (Doty) Doty ex P.C. Silva, a red macroalga, is a commercial source of carrageenan, a widely used polysaccharide compound important in the food and pharmaceutical industries, in nanotechnology, and in pharmacological applications. Carrageenan is found mainly in the cell wall and in the intercellular matrix. This is the first study to propose the characterization of carrageenans in vitro, using the auto-fluorescence properties of the alga treated with different polyamines: putrescine, spermidine, and spermine. This study suggests a four-phase cultivation sequence for seaweed farmers to enhance and assess the potential carrageenan yield of their crops. In phase 1, seedlings were treated with each of the polyamines. Explants were subsequently transferred through two additional culture phases before being planted on the sea farms in phase 4 and then harvested after 60 days for analysis. Images from transverse sections of 11 representative cultured K. alvarezii samples were obtained at 561 nm excitation wavelength for both the cell center and the cell wall of each sample. Spectral data were also analyzed using the spectral phasor algorithm of SimFCS developed at the Laboratory for Fluorescence Dynamics (www.lfd.uci.edu). We report on the identification of several spectral fluorescence emission fingerprints from different auto-fluorescence compounds spatially mapped using this technique. These fingerprints have the potential to improve strain selection of explants for enhanced carrageenan yield in seaweed farming operations as well as to enable wholesale pricing to correspond with crop quality.