A simple method for the freeze-preservation of zoospores of the green macroalga Enteromorpha intestinalis |
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Authors: | Rebecca Taylor Robert L Fletcher |
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Institution: | (1) Department of Biological Sciences, University of Dundee, DD1 4HN, UK;(2) Institute of Marine Sciences, University of Portsmouth, Ferry Road, Eastney, Portsmouth, Hampshire, PO4 9LY, UK |
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Abstract: | Settled zoospores of the green macroalga Enteromorpha intestinalis were subjected to several different freezing and storing
treatments at both cryogenic and non-cryogenic temperatures after which their viability was assessed using a spore germination
bioassay. Three different cooling rates were tested: slow cooling at –1°C min−1 and –0.5°C min−1 to end temperatures in the range –20°C to –40°C, and a two-step procedure whereby the spores were frozen to –30°C at a rate
of –1°C min−1 prior to immersion in liquid nitrogen at –196°C. Spore viability was also investigated using the cryoprotectants glycerol
and dimethyl suphoxide (DMSO), a reduced saline medium and various storage times. In the majority of experiments, the use
of a cryoprotectant during the freezing process significantly increased the viability of the spores, with DMSO affording slightly
greater protection than glycerol. All treatments produced high viabilities (ranging from 75.3–100.0%) after 5-min storage
at the different end temperatures. However, progressively longer storage up to 7 days generally resulted in a marked reduction
in viability. This was with the exception of spores frozen in a reduced saline medium; a medium of 75% seawater and either
5 or 10% DMSO greatly increased spore viability, with values of > 40% recorded for spores stored at –20°C for up to 5 weeks.
This revised version was published online in July 2006 with corrections to the Cover Date. |
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Keywords: | cryopreservation macroalgae Enteromorpha zoospores cooling rate cryoprotectant storage time |
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