title: Effects of ascorbic acid on PVS2 cryopreservation of Dendrobium Bobby Messina's PLBs supported with SEM analysis.
abstract: Regrowth of the cryopreserved protocorm-like bodies (PLBs) of Dendrobium Bobby Messina was assessed based on the plant vitrification solution 2 (PVS2) optimisation conditions. The optimized protocol obtained based on TTC spectrophotometrical analysis and growth recovery were 3-4 mm of PLBs size precultured in 0.2 M sucrose for 1 day, treated with a mixture of 2 M glycerol and 0.4 M sucrose supplemented with half-strength liquid MS media at 25 °C for 20 min and subsequently dehydrated with PVS2 at 0 °C for 20 min prior to storage in liquid nitrogen. Following rapid warming in a water bath at 40 °C for 90 s, PLBs were treated with unloading solution containing half-strength liquid MS media supplemented with 1.2 M sucrose. Subsequently, the PLBs were cultured on half-strength semi-solid MS media supplemented with 2 % (w/v) sucrose without any growth regulators and resulted in 40 % growth recovery. In addition, ascorbic acid treatment was used to evaluate the regeneration process of cryopreserved PLBs. However, growth recovery rates of non-cryopreserved and cryopreserved PLBs were 30 and 10 % when 0.6 mM ascorbic acid was added. Scanning electron microscopy analysis indicates that there are not much damages observed on both cryopreserved and non-cryopreserved PLBs in comparison to PLBs stock culture.
authors.affiliation: School of Biological Sciences, Universiti Sains Malaysia USM, Minden Heights 11800, Penang, Malaysia.
authors.forename: Jessica Jeyanthi James
authors.initials: JJ
authors.lastname: Antony
authors.forename: Chan Lai
authors.initials: CL
authors.lastname: Keng
authors.forename: Maziah
authors.initials: M
authors.lastname: Mahmood
authors.forename: Sreeramanan
authors.initials: S
authors.lastname: Subramaniam
mesh_terms: D001205:Ascorbic Acid / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D031667:Dendrobium / Q000166:cytology* / Q000187:drug effects* / Q000254:growth & development / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D064347:Microscopy, Electrochemical, Scanning; D013696:Temperature; D013997:Time Factors
chemical_list: D001205:Ascorbic Acid