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Cryopreservation of gametophytic thalli of Ulva prolifera (Ulvales, Chlorophyta) from Korea

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Abstract

Cryopreservation is needed to ensure long-term storage without any genetic drift and contamination. To conserve Ulva species, gametophytic thalli of Ulva prolifera (Chlorophyta) were first cryopreserved at −196 °C using the two-step cooling method with cooling rate of 1 °C min−1 to −40 °C. Three cryoprotectants (dimethyl sulfoxide (DMSO), glycerol, and proline) at five concentrations (5, 10, 15, 20, and 25 %) and mixed cryoprotectants were assessed to determine the optimal solution. The mixture of cryoprotectants with the highest viability was 10 % glycerol in combination with 5 % DMSO and 5 % proline, with a viability of 89.2 %. The cryoprotectant treated singly with the highest viability was 20 % glycerol with a viability of 91.6 %. However, there was no statistically significant difference between 20 % glycerol and the mixed treatment. An evaluation of long-term storage in liquid nitrogen showed very high viability of 91.2–92.1 % with no statistically significant effect from storage time up to 120 days. The specimens developed normally in culture after cryopreservation for 120 days with the rate of gametogenesis reaching 95.7 % on the fourth day of culture. The released gametes developed normally into gametothalli. In conclusion, the use of glycerol as a cryoprotectant was very effective for cryopreservation of the gametothalli of U. prolifera. With 20 % glycerol, viability was very high as 91.6 %. The results suggest that gametothalli of U. prolifera can be cryopreserved completely by a treatment with 15–25 % glycerol singly as well as by a mixture of cryoprotectants with usual efficiency as a cryoprotectant.

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Acknowledgments

This work was supported by a grant from Marine Biotechnology Program funded by Ministry of Oceans and Fisheries of Korean Government.

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  1. Department of Marine Biology, Pukyong National University, Busan, 608-737, Korea

    Ye Nee Lee & Ki Wan Nam

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  1. Ye Nee Lee
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  2. Ki Wan Nam
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Correspondence to Ki Wan Nam.

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Lee, Y.N., Nam, K.W. Cryopreservation of gametophytic thalli of Ulva prolifera (Ulvales, Chlorophyta) from Korea. J Appl Phycol 28, 1207–1213 (2016). https://doi.org/10.1007/s10811-015-0620-7

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  • DOI: https://doi.org/10.1007/s10811-015-0620-7

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