title: Cryopreservation of the gorgonian endosymbiont Symbiodinium.
abstract: The study focused on finding a suitable cryoprotectant (CPA) and an optimum freezing protocol for the cryopreservation of the endosymbiotic dinoflagellates (Symbiodinium, clade G) of Junceella fragilis wherein the success of experiments is crucial to both scientific and ecology studies. A two-step freezing technique was developed. The viability of the thawed dinoflagellates was assayed using the adenosine triphosphate (ATP) bioassay for the first time and was further confirmed through the culturing of dinoflagellates in vitro. The results suggested that 30 min was the most suitable holding time for the dinoflagellates, and the samples produced highest viability when suspended at 5 cm from the surface of LN2. Results also showed that 1 M methanol with 0.4 M sucrose was the most effective CPA, yielding the highest viability (56.93%). Although cell densities of both cryopreserved and control group suffered an initial decline of culture, the cell densities were maintained throughout the remaining duration. In the present study, the cryopreservation of clade G endosymbiont algae was studied for the first time and the method described here could be applied for future studies on symbiotic algae cryopreservation.
authors.affiliation: Institute of Marine Biology, National Dong Hwa University, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Chong
authors.affiliation: Department of Biotechnology, Mingdao University, 369 Wen-Hua Road, Peetow, ChangHua, 52345, Taiwan.|Department of Post Modern Agriculture, Mingdao University, 369 Wen-Hua Road, Peetow, Chang Hua, 52345, Taiwan.
authors.forename: Sujune
authors.initials: S
authors.lastname: Tsai
authors.affiliation: National museum of Marine Biology &Aquarium, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan.|Institute of Marine Biology, National Dong Hwa University, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan.
authors.forename: Li-Hsueh
authors.initials: LH
authors.lastname: Wang
authors.affiliation: Department of Aquaculture, National Taiwan Ocean University, 2 Beining Road, Jhongjheng, Keelung, Taiwan.
authors.forename: Chih-Yang
authors.initials: CY
authors.lastname: Huang
authors.affiliation: National museum of Marine Biology &Aquarium, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan.|Institute of Marine Biology, National Dong Hwa University, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan.
authors.forename: Chiahsin
authors.initials: C
authors.lastname: Lin
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002452:Cell Count; D015925:Cryopreservation; D003451:Cryoprotective Agents; D004141:Dinoflagellida / Q000502:physiology; D005615:Freezing; D013395:Sucrose; D013559:Symbiosis
chemical_list: D003451:Cryoprotective Agents; D013395:Sucrose; D000255:Adenosine Triphosphate