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Simultaneous time resolution of the emission spectra of fluorescent proteins and zooxanthellar chlorophyll in reef-building corals[para][dagger]
Photochemistry and Photobiology, May 2003 by Gilmore, Adam M, Larkum, Anthony W D, Salih, Anya, Itoh, Shigeru, Et al
Moreover, it is established from recent time-resolved fluorescence studies in vitro that oligomers comprising both immature green and mature red forms of the coral FP known as Discosoma Reel (Ds-Red) exhibit FRET (41,42). The reciprocal kinetic responses observed in vitro generally correspond with those obtained here in vivo with the blue and green FP. Notably, in comparison with one of the clearest case studies (41), our main decay component was faster for the blue-green acceptor
CONCLUSIONS
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This study demonstrates the use of in vivo simultaneous time and wavelength fluorescence acquisition and DCI analysis (27) for answering important physiological questions, namely, (1) that the blue and green FP do not function in light harvesting for reef-building corals that dwell in shallow water; and (2) that FRET is a primary process in the excited-state behavior of FP in reef-building corals. This study provides a foundation of data from nonstressed corals from key geographical regions of interest with respect to coral bleaching (13,44,45) based on which future studies of stress conditions may be initiated and compared. Future work should be aimed at exploring the mechanism of the proposed photoprotective function of the FP and the physiological state of both the FP and zooxanthellae in response to coral-bleaching episodes. The fluorescence lifetimes and spectra should provide a valid quantitative indicator, especially under conditions where the tissue concentrations and chemical properties of either zooxanthellae or FP are likely to change simultaneously. This is because determination of the fluorescence intensities and ratios alone is complicated by concomitant changes in the concentration and excited-state lifetimes of fluorophores in the tissue. Finally, the methods used for identification of the new FP and FRET partners described in this study may be of interest to researchers who wish to isolate, clone and mutate FP for molecular technology, as is becoming increasingly common for the green and red FP from other cnidarians.
Acknowledgements-A.M.G. and S.I. thank the Japanese Society for the Promotion of Science for an invited visiting fellowship to A.M.G. Special thanks are offered to Dr. Mino Hiroyuki and Kana Sugiura in Nagoya, Japan, and Drs. Yasuko Sakihama and Shunichi Takahashi and Takashi Nakamura, M.Sc., in Okinawa, Japan, for experimental assistance with coral collection, shipment and storage. R.V.W. and H.Y. were supported by a Grant-in Aid for Scientific Research (B) (12480166) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan. A.M.G. thanks Dr. Marilyn Ball and the Ecosystem Dynamics group of the ANU RSBS for stimulating discussions.
[para]Posted on the website on 6 March 2003.
[dagger]The results of this study were presented in their entirety as a poster at the ComBio2002 conference held in Sydney, Australia, 29 September-3 October 2002.
REFERENCES
1. Kawaguti, S. (1944) On the physiology of reef corals. VI. Study on the pigments. Palao Trop. Biol. Stn. Stud. 2, 617-674.
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