ScienceAsia 33 (2007): 223-227|doi: 10.2306/scienceasia1513-1874.2007.33.223

Cryopreservation of Vanda coerulea Griff. ex Lindl. Seeds by Vitrification

Kanchit Thammasiri* and Luxsana Soamkul

 
ABSTRACT: An effective protocol for cryopreservation of seeds of Vanda coerulea Griff. ex Lindl., an endangered Thai epiphytic orchid, in liquid nitrogen (LN) was developed by vitrification. Seeds from 7-month-old pods were sufficiently dehydrated in 2 ml cryotubes filled with highly concentrated plant vitrification solution (PVS2), consisting of 30% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) dimethyl sulfoxide, prepared in modified Vacin and Went (1949) (VW) liquid medium at 25 ± 2°C for 70 min. The seeds were then rapidly plunged into LN. After recovering from LN and rapid warming in a waterbath at 40°C for 2 min, the PVS2 solution was replaced with 0.5 ml of 1.2 M sucrose in VW solution and kept at 25 ± 2°C for 20 min prior to transfer on VW agar medium. About 67% of cryopreserved seeds treated with PVS2 solution for 70 min were able to develop into normal seedlings in vitro, while without PVS2 treatment, there was no survival after LN storage.


KEYWORDS: Vanda coerulea Griff. ex Lindl., Thai orchid, orchid seeds, vitrification, cryopreservation.

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