Abstract
Information on the ecophysiology of macroalgal planktonic propagules (e.g. spores) has been hard to obtain, given their small size and low concentration in the water column. Studies of the photo-physiology of macroalgal spores, for example, have been limited by the need to aggregate many spores into bulk samples for analysis. Subsequently, physiological variability among spores (e.g. pigment concentration, absorption characteristics) is lost, and taxonomic comparisons from multi-taxa samples are impossible. Here we present a technique that utilizes a spectral microphotometer to produce visible (400-800 nm) absorption spectra from individual particles; the particles in our case are macroalgal spores. The microphotometer consists of a microscope fitted with a monochromator and spectrophotometer. After mounting spores from laboratory or field suspensions onto transparent membrane filters, absorption characteristics of individual spores, or even individual plastids, can be evaluate d independently from the remaining particles in the sample. Use of transparent rather than opaque membrane filters allows for determination of absorption spectra, as well as more traditional microscopic analyses (e.g. bright field, dark field, epi-fluorescence). Glutaraldehyde fixation and cold storage (−10 °C) were found to be appropriate for maintaining the integrity of absorption spectra for at least 3 days. To demonstrate the utility of microphotometry for macroalgal studies, absorption spectra were obtained and analyzed from spores of various kelps and filamentous red algae.
Similar content being viewed by others
References
Amsler, C. D. & M. Neushul, 1991. Photosynthetic physiology and chemical composition of spores of the kelps Macrocystis pyrifera, Nereocystis luetkeana, Laminaria farlowii, and Pterygophora californica(Phaeophyceae). J. Phycol. 27: 26–34.
Beach, K. S., C. M. Smith, T. Michael & H. Shin, 1995. Photosynthesis in reproductive unicells of Ulva fasciataand Enteromorpha flexuosa: implications for ecological success. Mar. Ecol. Prog. Ser. 125: 229–237.
Bidigare, R. R., M. E. Ondrusek, M. C. Kennicutt II, R. Iturriaga, H. R. Harvey, R. R. Hoham & S. A. Macko, 1993. Evidence for a photoprotective function for secondary carotenoids of snow algae. J. Phycol. 29: 427–434.
Brzezinski, M. A., D. C. Reed & C. D. Amsler, 1993. Neutral lipids as major storage products in zoospores of the giant kelp Macrocystis pyrifera(Phaeophyceae). J. Phycol. 29: 16–23.
Carpenter, E. J., J. Chang & L. P. Shapiro, 1991. Green and blue fluorescing dinoflagellates in Bahamian waters. Mar. Biol. 108: 145–149.
Carpenter, E. J., J. Chang, M. Cotrell, J. Schubauer, H. W. Paerl, B. M. Bebout & D. G. Capone, 1990. Re-evaluation of nitrogenase oxygen-protective mechanism in the planktonic marine cyanobacterium Trichodesmium. Mar. Ecol. Prog. Ser. 65: 151–158.
Garbary, D. J., K. Y. Kim, T. Klinger & D. Duggins, 1999. Preliminary observations on the development of kelp gametophytes endophytic in red algae. Hydrobiologia, 398/399 (Dev. Hydrobiol. 137): 247–252.
Grzymski, J., G. Johnsen & E. Sakshaug, 1997. The significance of intracellular self-shading on the biooptical properties of brown, red, and green macroalgae. J. Phycol. 33: 408–414.
Henry, E. C. & K. M. Cole, 1982. Ultrastructure of swarmers in the Laminariales (Phaeophyceae). I. Zoospores. J. Phycol. 18: 550–569.
Iturriaga, R. & D. A. Siegel, 1989. Microphotometric characterization of phytoplankton and detrital absorption properties of the Sargasso Sea. Limnol. Oceanogr. 34: 1706–1726.
Iturriaga, R., B. G. Mitchell & D. A. Kiefer, 1988. Microphotometric analysis of individual particle absorption spectra. Limnol. Oceanogr. 33: 128–135.
Jeffrey, S.W., R. F. C. Mantoura & S.W. Wright, 1997. Phytoplankton pigments in oceanography: guidelines to modern methods. UNESCO Publishing, Paris, 661 pp.
Lobban, C. S. & P. J. Harrison, 1994. Seaweed ecology and physiology. Cambridge University Press, Cambridge, 366 pp.
Madden, H. H., 1978. Comments on the Savitsky-Golay convolution method for least-squares fit smoothing and differentiation of digital data. Anal. Chem. 50: 1383–1386.
Robinson, D. H., K. R. Arrigo, R. Iturriaga & C. W. Sullivan, 1995. Microalgal light-harvesting in extreme low-light environments in McMurdo Sound, Antarctica. J. Phycol. 31: 508–520.
Rowan, K. S., 1989. Photosynthetic Pigments of Algae. Cambridge University Press, Cambridge, 334 pp.
Savitsky, A. & M. J. E. Golay, 1964. Smoothing and differentiation of data by simplified least squares procedures. Anal. Chem. 36: 1627–1639.
Smith, C. M. & R. S. Alberte, 1994. Characterization of in vivoabsorption features of chlorophyte, phaeophyte and rhodophyte algal species. Mar. Biol. 118: 511–521.
Steinier, J., Y. Termonia & J. Deltour, 1972. Comments on smoothing and differentiation of data by simplified least square procedures. Anal. Chem. 44: 1906–1909.
Stephens, F. C., 1995. Variability of spectral absorption efficiency within living cells of Pyrocystis lunula(Dinophyta). Mar. Biol. 122: 325–331.
Troussellier, M., C. Courties & S. Zettelmaier, 1995. Flow cytometric analysis of coastal lagoon bacterioplankton and picophytoplankton: fixation and storage effects. Estuar. coast. shelf Sci. 40: 621–633.
Underwood, A. J., 1997. Experiments in ecology: their logical design and interpretation using analysis of variance. Cambridge University Press, Cambridge, 504 pp.
Vaulot, D., C. Courties & F. Partensky, 1989. A simple method to preserve oceanic phytoplankton for flow cytometric analysis. Cytometry 10: 629–635.
Wilson, P. D. & S. R. Polo, 1981. Polynomial filters of any degree. J. opt. Soc. Am. 71: 599–603.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Graham, M.H., Mitchell, B.G. Obtaining absorption spectra from individual macroalgal spores using microphotometry. Hydrobiologia 398, 231–239 (1999). https://doi.org/10.1023/A:1017009411367
Issue Date:
DOI: https://doi.org/10.1023/A:1017009411367