Effects of droplet-vitrification on Brassidium Shooting Star’s orchid protocorm-like bodies (PLBs)

摘要: Abstract In the present study, protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid was successfully cryopreserved using droplet-vitrification method. Seven (7) parameters have been optimised such as PLB size, sugar type, preculture concentration, incubation duration, plant vitrification solution 2 (PVS2) thawing method and unloading duration. Assessments viability were evaluated by TTC spectrophotometric at 490 nm absorbance growth recovery regenerated PLBs. The results showed that survival plants superior when 3–4 mm PLBs precultured on sucrose enriched media. Decrease in observed exposed to media higher than 0.25 M sucrose. However, extended duration 7 days improved non-cryopreserved 70%. study proved PVS2 treatment promotes cryopreseved Regenerated treated with for 40 min before plunging into liquid nitrogen. highest 30% obtained semi-solid supplemented days, followed loading 20 min, dehydration prior store nitrogen rinsing 15 min. Addition ascorbic acid α-tocopherol which reported regrowth several did not affect orchid’s inclusion preculture, loading, steps enhances faster three weeks after treatment.