Abstract
Chitinases are involved in defense against chitinaceous pathogens in both invertebrates and vertebrates. This study investigated whether sea fan corals, Gorgonia ventalina (Linnaeus) collected from the Florida Keys between the summer of 2002 and the summer of 2005 contain chitinases, and whether these enzymes could serve an analogous protective role against the fungal pathogen, Aspergillus sydowii (Thom et Church). Crude extracts of healthy sea fans contained detectible levels of exochitinase activity in an in vitro microplate assay using fluorogenic substrates. The exochitinase levels decreased upon injury, agitation, or manipulation of the tissue. A concurrent, transient increase of exochitinase in the surrounding water suggests that sea fans release chitinases as a response to these stresses. By contrast, endochitinase was detected in only 2 of 15 sea fans (13%), suggesting a high degree of variation for this enzyme. Sea fan chitinase-containing seawater and anion exchange chromatography fractions were both active against A. sydowii in an absorbance-based antifungal assay. The presence of chitinases in sea fan extracts, their release into the surrounding water upon stress, and their activity against A. sydowii suggests that further study of these enzymes in coral stress responses is warranted.
Similar content being viewed by others
References
Agrawal AA, Tuzun S, Bent E (eds) (1999) Induced plant defenses against pathogens and herbivores: biochemistry, ecology, and agriculture. The American Phytopathological Society Press, St. Paul, Minnesota
Alker A, Kim K, Dube D, Harvell CD (2004) Localized induction of a generalized response against multiple biotic agents in Caribbean sea fans. Coral Reefs 23:397–405
Berenbaum MR, Zangerl AR (1999) Coping with life as a menu option: inducible defenses of the wild parsnip. In: Tollrian R, Harvell CD (eds) The ecology and evolution of inducible defenses. Princeton University Press, Princeton, New Jersey, pp 10–32
Brown BE, Bythell JC (2005) Perspectives on mucus secretion in reef corals. Mar Ecol Prog Ser 296:291–309
Dixon DM, Walsh TJ (1992) Human pathogenesis. Biotechnology 23:249–267
Elyakova LA (1972) Distribution of cellulases and chitinases in marine invertebrates. Comp Biochem Physiol 43:67–70
Flach J, Pilet PE, Jolles P (1992) What’s new in chitinase research? Experientia 48:701–716
Geffen Y, Rosenberg E (2005) Stress-induced rapid release of antibacterials by scleractinian corals. Mar Biol 146:931–935
Geiser DM, Taylor JW, Ritchie KB, Smith GW (1998) Cause of sea fan death in the West Indies. Nature 394:137–138
Gil-Agudelo DL, Myers C, Smith GW, Kim K (2006) Changes in the microbial communities associated with Gorgonia ventalina during Aspergillosis infection. Dis Aquat Org 69:89–94
Gillespie JP, Kanost MR, Trenczek T (1997) Biological mediators of insect immunity. Ann Rev Entomol 42:611–43
Jensen PR, Harvell CD, Wirtz K, Fenical W (1996) The incidence of anti-microbial activity among Caribbean gorgonians. Mar Biol 125:411–420
Kim K, Harvell CD, Kim PD, Smith GW, Merkel SM (2000a) Fungal disease resistance of Caribbean sea fan corals (Gorgonia spp.). Mar Biol 136:259–267
Kim K, Kim PD, Alker AP, Harvell CD (2000b) Antifungal properties of gorgonian corals. Mar Biol 137:393–401
Klug M, Tardent P, Smid I, Holstein T (1984) Presence and localization of chitinase in Hydra and Podocoryne (Cnidaria, Hydrozoa). J Exp Zool 229:69–72
Knowlton N, Rohwer F (2003) Multispecies microbial mutualisms on coral reefs: the host as a habitat. Am Nat 162:S51–S62
Kramer KJ, Muthukrishnan S (1997) Insect chitinases: molecular biology and potential use as biopesticides. Insect Biochem Molec 27:887–900
Levin DA (1976) The chemical defenses of plants to pathogens and herbivores. Annu Rec Ecol Syst 7: 121-159
Mali B, Mohrlen F, Frohme M, Frank U (2004) A putative double role of a chitinase in a cnidarian: pattern formation and immunity. Dev Comp Immunol 28:973–981
Merzendorfer H, Zimoch L (2003) Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases. J Exp Biol 206:4393–4412
Nagelkerken I, Buchan K, Smith GW, Bonair K, Bush P, Garzón-Ferreira J, Botero L, Gayle P, Harvell CD, Heberer C, Kim K, Petrovic C, Pors L, Yoshioka P (1997) Widespread disease in Caribbean sea fans: II. Patterns of infection and tissue loss. Mar Ecol Prog Ser 160:255–263
Nagy NE, Fossdal CG, Dalen LS, Lonneborg A, Heldal I, Johnsen O (2004) Effects of Rhizoctonia infection and drought on peroxidase and chitinase activity in Norway spruce (Picea abies). Physiol Plant 120:465–473
Ng TB (2004) Antifungal proteins and peptides of leguminous and non-leguminous origins. Peptides 25:1215–1222
Olano CT, Bigger CH (2000) Phagocytic activities of the gorgonian coral Swiftia exserta. J Invertebr Pathol 76:176–184
Overdijk B, Van Steijn GJ, Odds FC (1999) Distribution of chitinase in guinea pig tissues and increases in levels of this enzyme after systemic infection with Aspergillus fumigatus. Microbiology 145:259–269
Petes L, Harvell CD, Peters EC, Webb MAH, Mullen KM (2003) Pathogens compromise reproduction and induce melanization in Caribbean sea fans. Mar Ecol Prog Ser 264:167–171
Ritchie KB, Smith GW (2004) Microbial communities of coral surface mucopolysaccharide layers. In: Rosenburg E, Loya Y (eds) Coral health and disease. Springer, Berlin Heidelberg New york, pp 259–263
Rohwer F, Seguritan V, Azam F, Knowlton N (2002) Diversity and distribution of coral-associated bacteria. Mar Ecol Prog Ser 243:1–10
Shibuya K, Naoe S, Yamaguchi H (1999) Animal models of A. fumigatus infections. Contrib Microbiol 2:130–138
Slattery M, Hamann MT, McClintock JB, Perry TL, Puglisi MP, Yoshida WY (1997) Ecological roles for water-borne metabolites from Antarctic soft corals. Mar Ecol Prog Ser 161:133–144
Smith GW, Ives LD, Nagelkerken IA, Ritchie KB (1996) Caribbean sea fan mortalities. Nature 383:487
Suzuki M, Morimatsu M, Yamashita T, Iwanaga T, Syuto B (2001) A novel serum chitinase that is expressed in bovine liver. FEBS Lett 506:127–130
Thevissen K, Osborn RW, Acland DP, Broekaert WF (2000) Specific binding sites for an antifungal plant defensin from Dahlia (Dahlia merckii) on fungal cells are required for antifungal activity. Mol Plant Microbe Interact 13:54–61
Trench RK (1996) Specificity and dynamics of algal–invertebrate symbiosis. Symbiosis 96:16
Tronsmo A, Harman GE (1993) Detection and quantification of N-acetyl-beta-d-glucosaminidase, chitobiosidase, and endochitinase in solutions and on gels. Anal Biochem 208:74–79
Tuzun S, Bent E (1999) The role of hydrolytic enzymes in multigenic and microbially-induced resistance in plants. In: Agrawal AA, Tuzun S, Bent E (eds) Induced plant defenses against pathogens and herbivores. APS Press, St Paul, pp 95–115
VanEtten HD, Mansfield JW, Bailey JA, Farmer EE (1994) Two classes of plant antibiotics: phytoalexins versus phytoanticipins. Plant Cell 6:1191–1192
Acknowledgments
We thank K. Boor and J. Thaler for use of their microplate readers and we thank K. Rypien, J. Ward, and L. Mydlarz for critical reading. We also thank the staff of the Mote Marine Laboratories, Tropical Research Lab for their help in our experiments at their facility. This work was conducted under National Science Foundation grant number OCE-0326705 and collections were made under Florida Keys National Marine Sanctuary Research permit number FKNMS-2004-092.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by J.P. Grassle, New Brunswick.
Rights and permissions
About this article
Cite this article
Douglas, N.L., Mullen, K.M., Talmage, S.C. et al. Exploring the role of chitinolytic enzymes in the sea fan coral, Gorgonia ventalina . Mar Biol 150, 1137–1144 (2007). https://doi.org/10.1007/s00227-006-0444-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00227-006-0444-8