Optimization of a Cryopreservation Method for the Endangered Korean Species Pogostemon yatabeanus Using a Systematic Approach: The Key Role of Ammonium and Growth Regulators

Plants

Hyo-Eun Lee¹  Haeng-Hoon Kim¹  Ha-Na Park¹  Sang-Un Park²  Elena Popova³ 

[1] Department of Agricultural Life Science, Sunchon National University, Suncheon 57922, Korea;Division of Plant Science and Resources, Chungnam National University, Daejeon 34134, Korea;K.A. Timiryazev Institute of Plant Physiology of Russian Academy of Sciences, 127276 Moscow, Russia;

关键词: alternative vitrification solutions;    ammonium stress;    droplet-vitrification;    endangered species conservation;    osmoprotection;    plant growth regulators;   

DOI  :  10.3390/plants10102018

来源: DOAJ

【 摘 要 】

Cryopreservation provides a secure long-term conservation option for rare and endangered plant species with non-orthodox or limitedly available seeds. Wide application of cryopreservation to biobank wild flora is hampered by the need to re-optimize nearly all protocol steps for every new species. We applied a systematic approach to simplify optimization of a multi-stage droplet-vitrification method for the endangered wetland Korean species, Pogostemon yatabeanus. This approach consisted of a standard procedure pre-selected based on material type and size, which was complemented with 11 additional treatments to reveal the most impactful conditions. Effect of ammonium nitrate at various protocol steps was also tested. The highest shoot tip survival (92%) and plant regeneration (90%) after cryopreservation were achieved using preculture with 10% sucrose followed by 40 min osmoprotection and 60 min treatment with vitrification solution A3-80% (33.3% glycerol + 13.3% dimethyl sulfoxide + 13.3% ethylene glycol + 20.1% sucrose) on ice. A three-step regrowth procedure starting with ammonium-free medium with 1 mg/L GA₃ and 1 mg/L BA followed by ammonium-containing medium with and without growth regulators was essential for the development of healthy plants from cryopreserved shoot tips. This approach enables fast optimization of the cryopreservation procedure for new osmotic stress-sensitive plant species.

【 授权许可】

Unknown