Cryopreservation of Sporothalli of the Genus Porphyra (Bangiales, Rhodophyta) from Kor

doi:2003.18.4.321

Algae > Volume 18(4); 2003 > Article

Algae 2003;18(4): 321-331. doi: https://doi.org/10.4490/ALGAE.2003.18.4.321

Young-Hyun Jo, Sung-Pil Kang, Tae-Ho Seo, Sung-Je Choi, Kang Hee Kho, Kazuyoshi Kuwano, Naotsune Saga, Min-Yong Kim, Jong-Ahm Shin

Jeollannamdo Fisheries Research Institute, Division of Aqualife Science, Yosu National University, Koheung Fisheries Management Division, Yeosu Regional Maritime Affairs and Fisheries Office, Graduate School of Science and Technology, Nagasaki University, Graduate School of Fisheries Science, Hokkaido University, Department of Regrigeration Engineering, Yosu National University

ABSTRACT

Cryopreservation of sporothalli of the red alga Porphyra (P. seriata, P. yezoensis, P. tenera, P. pseudolinearis and P. dentata) was performed by the two-step cooling method in liquid nitrogen. The algal samples were suspended in various cryoprotective solutions, and slowly cooled to -40℃ in 4 hours using a programmed freezer. After the first slow cooling the suspensions with cryoprotectants were immediately immersed in liquid nitrogen. The suspension from the programmed freezer was thawed quickly by agitation of the vial in a water bath at 40℃. When ice in the suspension of cryogenic vial was mostly melted, the vial was transfered to an ice bath for complete melting of the residual ice. The cryoprotectants in the vial were washed off by gradual dilution with seawater. The viability of the cell was assessed with neutral red staining. The viability of Porphyra samples ranged 54.6-70.9% when the mixed suspension of 10% dimethylsulfoxide and 0.5M sorbitol in 50% seawater used as a cryoprotectant.

Key words: cryopreservation, Porphyra, sporothallus, two-step cooling method