The chemical structures of ultraviolet (UV)- absorbing substances secreted from the healthy living brown algae, Eisenia bicyclis and Ecklonia kurome, were demonstrated. The living activity of algal cells was critically examined using a confocal laser-scanning microscope after incorporation of fluorescein diacetate (FDA) into the cells. Using thin-layer chromatography (TLC), reversed-phase three-dimensional high-performance liquid chromatography (RP-3D-HPLC) and gas chromatography-mass spectrometry (GC-MS), it was found that the UV-absorbing substances (λmax 265–270 nm) secreted from the living brown algae mainly consisted of the three monomeric bromophenols, 2,4-dibromophenol, 2,4,6-tribromophenol and dibromo-iodophenol, but not phloroglucinol or phlorotannins. The other minor compounds detected in the secretions were as follows: benzothiazole, fatty acids (14:1, 16:0 and 18:0 acids), franesol, 3-hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate and squalene. Exudation of phloroglucinol and phlorotannins was ascertained to take place only after the cell death of these algae. These results indicate that, whilst the algae are alive, polymeric phlorotannins are strictly kept within the algal body, and only monomeric bromophenols are secreted into the seawater medium.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
gas chromatography-mass spectrometry
reversed-phase three-dimensional high-performance liquid chromatography
Bailey RW, Bourne EJ (1960) Colour reactions given by sugars and diphenylamine-aniline spray reagents on paper chromatograms. J Chromatog 4:206–213
Carlson DJ (1982) Surface microlayer phenolic enrichments indicate sea surface slicks. Nature 296:426–429
Carlson DJ, Carlson ML (1984) Reassessment of exudation by fucoid macroalgae. Limnol Oceanogr 29:1077–1087
Craigie JS, McLachlan J (1964) Excretion of colored ultraviolet-absorbing substances by marine algae. Can J Bot 42:23–33
Fogg GE, Boalch GT (1958) Extracellular products in pure cultures of a brown alga. Nature 181:789–790
Folch J, Less M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226:497–509
Jennings JG, Steinberg PD (1994) In situ exudation of phlorotannins by the sublittoral kelp Ecklonia radiata. Mar Biol 121:349–354
Krebs KG, Heusser D, Wimmer H (1969) Spray reagents. In: Stahl E (ed.) Thin-layer chromatography. Springer-Verlag, Heidelberg pp 854–909
McLachlan J, Craigie, JS (1964) Algal inhibition by yellow ultraviolet-absorbing substances from Fucus vesiculosus. Can J Bot 42:287–292
Nagayama K, Iwamura Y, Shibata T, Hirayama I, Nakamura T (2002) Bactericidal activity of phlorotannins from the brown alga Ecklonia kurome. J Antimicrob Chemother 50:889–893
Nagayama K, Shibata T, Fujimoto K, Honjo T, Nakamura T (2003) Algicidal effect of phlorotannins from the brown alga Ecklonia kurome on red tide microalgae. Aquaculture 218:601–611
Nakamura T, Nagayama K, Uchida K, Tanaka R (1996) Antioxidant activity of phlorotannins isolated from the brown alga Eisenia bicyclis. Fisheries Sci 62:923–926
Pedersen M, Collen J, Abrahamsson K, Ekdahl A (1996) Production of halocarbons from seaweeds: an oxidative stress reaction?. Sci Mar 60 (supl. 1): 257–263
Pellegrini L (1980) Cytological studies on physodes in the vegetative cells of Cystoseira stricta Sauvageau (Phaeophyta, Fucales). J Cell Sci 41:209–231
Ragan MA, Glombitza K-W (1986) Phlorotannins, brown algal polyphenols. Progr Phycol Res 4:129–241
Sakami T (1996) Effects of algal excreted substances on the respiration activities of epiphytic bacteria on the brown alga, Eisenia bicyclis Kjellman. Fisheries Sci 62:394–396
Shibata T, Yamaguchi K, Nagayama K, Kawaguchi S, Nakamura T (2002a) Inhibitory activity of brown algal phlorotannins against glycosidases from the viscera of the turban shell Turbo cornutus. Eur J Phycol 37: 493–500
Shibata T, Fujimoto K, Nagayama K, Yamaguchi K, Nakamura T (2002b) Inhibitory activity of brown algal phlorotannins against hyaluronidase. Int J Food Sci Tech 37:703–709
Shibata T, Nagayama K, Tanaka R, Yamaguchi K, Nakamura T (2003) Inhibitory effects of brown algal phlorotannins on secretory phospholipase A2s, lipoxygenases and cyclooxygenases. J Appl Phycol 15:61–66
Shibata T, Kawaguchi S, Hama Y, Inagaki M, Yamaguchi K, Nakamura T (2004) Local and chemical distribution of phlorotannins in brown algae. J Appl Phycol 16:291–296
Sieburth J McN, Conover JT (1965) Sargassum tannin, an antibiotic which retards fouling. Nature 208:52–53
Swanson AK, Druehl LD (2002) Induction, exudation and the UV protective role of kelp phlorotannins. Aquat Bot 73:241–253
Taniguchi K, Akimoto Y, Kurata K, Suzuki M (1992) Chemical defense mechanism of the brown alga Eisenia bicyclis against marine herbivores. Nippon Suisan Gakkaishi 58:571–575
Tatewaki M (1966) Formation of a crustaceous sporophyte with unilocular sporangia in Scytosiphon lomentaria. Phycologia 6:62–66
Widholm JM (1972) The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol 47:189–194
About this article
Cite this article
Shibata, T., Hama, Y., Miyasaki, T. et al. Extracellular secretion of phenolic substances from living brown algae. J Appl Phycol 18, 787–794 (2006). https://doi.org/10.1007/s10811-006-9094-y
- Ecklonia kurome
- Eisenia bicyclis
- Gas chromatography-mass spectrometry
- Thin-layer chromatography