Thai Journal of Botany
Article's details
Title: Cryopreservation of Alpinia nigra (Gaertn.) Burtt by Vitrifi cation-based technique
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Pages: 151 - 154
Year: 2553
Year No.: 2
Volume: พิเศษ   Show All Articles
Abstract:         In the cryopreservation of Alpinia nigra (Gaertn.) Burtt, shoot tips from in vitro plantlets cultured on Murashige and Skoog (1962) (MS) agar medium for 1 month were dehydrated with a preculture solution (0.3 M sucrose) for 3 days on a rotary shaker at 100 rpm and subsequently exposed to a cryoprotectant solution (5% w/v DMSO and 5% w/v glycerol) and a loading solution (0.4 M sucrose and 2 M glycerol) for 20 min in each solution. They were kept in 1.8 ml cryotubes, containing PVS2 solution [0.4 M sucrose, 15% (w/v) ethylene glycol, 15% (w/v) DMSO and 30% (w/v) glycerol] for 0, 20, 40, 60, 80 or 100 min, respectively, before being immediately plunged into liquid nitrogen for 1 hour. Thereafter, cryotubes were taken out of a liquid nitrogen tank and rapidly warmed in a waterbath at 40°C for 2 min. PVS2 solution was replaced with an unloading solution (1.2 M sucrose) for 20 min before shoot tips were cultured on MS agar medium supplemented with 1 ppm TDZ for 1 month and subsequently transferred to PGRs free MS agar medium for 1 month. The shoot tips exposed to PVS2 solution for 80 min showed the highest percentage of shoot and root development at 27.
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