Cryopreservation of protocorm-like body-derived shoot tips of Calanthe davidii by droplet-vitrification.
キーワード
概要
BACKGROUND
Evaluation of the conservation status of Calanthe plants in China indicated that 18 species were endangered or critically endangered. Comprehensive conservation solutions including ex situ methods, are urgently required to protect Calanthe species in China.
OBJECTIVE
The study aims to develop a simple and efficient cryopreservation protocol using droplet-vitrification for Calanthe davidii.
METHODS
Protocorm-like bodies (PLBs) were induced from nodal sections of in vitro shoots, and their proliferation was promoted by a thin cell layer culture procedure. Shoot tips excised from three leaf-stage PLBs were used in cryopreservation experiments. Key factors of the droplet-vitrification procedure including sucrose preculture, treatment with PVS2 solution and post-rewarming culture conditions were optimized to achieve a high level of regeneration.
RESULTS
When the optimized procedure was applied, 77.8 ± 3.9% of cryopreserved shoot tips withstood liquid nitrogen exposure and regenerated into new PLBs.
CONCLUSIONS
These results highlighted the importance of post-rewarming osmo-conditioning for regeneration of cryopreserved shoot tips.
