Effect of cryopreservation on proteins from the ubiquitous marine dinoflagellate Symbiodinium sp.

• Main Authors: Jia-Lin Lu, 呂佳琳
• Other Authors: Chia-Hsin Lin
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/9pn566

碩士 === 國立東華大學 === 海洋生物研究所 === 107 === Symbiotic algae play an important role in coral reef ecosystems. There is an inter-symbiotic relationship between coral and algae by providing energy to coral through photosynthesis process. Therefore, Symbiotic algae play an important role in coral reef ecosystems. Cryopreservation become an indispensable tool for species preserving and gene bank establishment, however this technology for symbiotic algae not yet matured and there is no study on the molecular level of symbiotic algae after thawing. Hence it is important to establish a suitable freezing protocol as well as to understand molecular impact by cryo-injury of symbiotic algae. The objective of this study was to identify suitable freezing conditions including the use of cryoprotectants (CPAs) and equilibration time through two-step freezing technique with a cooling rate of -59.83°C/for clade B Symbiodinium which were culture for 3, 7, 14 and 28 days after freezing and thawing. Then LHP and RFP were studied by western blotting for protein expression analysis while Adenosine triphosphate bioassay (ATP) for cell viability. The result showed that 2M propylene glycol (PG) with 30 minute equilibration time and 2M methanol (MeOH) with 20 minute equilibration time had best viabilities after thawing. Both treatments showed significantly increased growth rate after 28 days culture especially for the treatment by MeOH which was twice higher compared to PG treatment. The total amount of proteins is affected by low temperature. MeOH treatment did not show any significant difference from control, howeverin PG treatment group, it showed increasing protein expression after 28 days of culture. This study provides better understanding on freezing protocol for Symbiodinium clade B and the effects of low temperature towards protein expression in symbiotic algae.