Visualization of plastids and lipophilic components in living colonies of a wild strain of the hydrocarbon-forming, green alga Botryococcus by laser scanning confocal microscopy |
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Authors: | Gordon W Beakes Ann L Cleary |
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Institution: | (1) Plant Cell Biology Group, Research School of Biological Sciences, Australian National University, Canberra, ACT, 2801, Australia;(2) Department of Biological and Nutritional Sciences, The University, Newcastle upon Tyne, NE1 7RU, UK |
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Abstract: | Colonies of a wild strain from Lake Burley-Griffin, Australia, of the hydrocarbon-producing green alga Botryococcus were examined
by confocal laser scanning microscopy. The microscope was fitted with a dual wavelength krypton-argon laser, which permitted
simultaneous detection of chlorophyll autofluorescence and lipophilic dye fluorescence. This quick and simple technique revealed
the precise structural conformation of the autofluorescing plastids in living cells and their 3-dimensional spatial arrangement
within the dense globular colonies. Cells stained with the lipophilic carbocyanine dye, DIOC6(3) contain an apical array of
intensely staining granules as well as a more diffuse internal cisternal system thought to be endoplasmic reticulum. The cationic
lipophilic dye rhodamine123 revealed a finer reticulate system in the outermost cytoplasm partially overlaying the plastid.
Both dyes revealed the lipophilic nature of the extracellular matrix and enabled the secretion of lipid globules exuded from
the colonies to be visualized. It is suggested that confocal laser scanning microcopy would make an ideal tool to screen isolates
for their potential to form and secrete hydrocarbon, processes which are still far from clearly understood in this potentially
commercially important alga.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | Botryococcus plastid morphology hydrocarbon localization Confocal Laser Scanning Microscopy DIOC6(3) rhodamine123 |
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