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Zheng Ling-Ling, Zhang Qi, Li Tian-Li, Song Li-Rong. Effect and optimization of cryopreservation on vegetative cells and akinete of Haematococcus pluvialis Flotow FACHB-712[J]. Plant Science Journal, 2017, 35(5): 767-773. DOI: 10.11913/PSJ.2095-0837.2017.50767
Citation: Zheng Ling-Ling, Zhang Qi, Li Tian-Li, Song Li-Rong. Effect and optimization of cryopreservation on vegetative cells and akinete of Haematococcus pluvialis Flotow FACHB-712[J]. Plant Science Journal, 2017, 35(5): 767-773. DOI: 10.11913/PSJ.2095-0837.2017.50767

Effect and optimization of cryopreservation on vegetative cells and akinete of Haematococcus pluvialis Flotow FACHB-712

  • The survival rate of cryopreservation based on vegetative cells and akinetes of Haematococcus pluvialis Flotow FACHB-712 were compared. Results showed that in most cases the survival rate of the vegetative cells was lower than 5% in liquid nitrogen with two-step cryopreservation, but the rate increased to 13.3% if the protocol was modified as follows:cell suspensions were added with glycerol as a cryoprotectant (10%), cooled at a rate of 0.5℃/min until -40℃, and then maintained for 30 min before being placed to liquid nitrogen. The survival rate of the akinetes reached 66.13% in liquid nitrogen with the two-step protocol. There were no obvious differences (P > 0.05) in the growth characteristics and the astaxanthin content of akinetes before and after preservation in liquid nitrogen. Comparatively, the ultrastructure of the vegetative cells was seriously damaged, whereas that of the akinetes was much less damaged after two-step cryopreservation treatment. If directly placed in -20℃, -80℃, or liquid nitrogen, respectively, the highest survival rate of the samples was about 27% in the treatment under -80℃. In conclusion, the two-step cryopreservation protocol in liquid nitrogen using akinete-stage cells was preferable for the long-term preservation of H. pluvialis FACHB-712, with the one-step protocol at -80℃ also applicable as an alternative approach.
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