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Asian Journal of Plant Sciences
Year: 2007  |  Volume: 6  |  Issue: 8  |  Page No.: 1235 - 1240

In vitro Conservation of Cleisostoma areitinum (Rchb.f.) Garay, Rare Thai Orchid Species by an Encapsulation-Dehydration Method

P. Maneerattanarungroj, S. Bunnag and M. Monthatong    

Abstract: Protocorm cryopreservation has been performed for ex situ preservation of Cleisostoma areitinum (Rchb.f.) Garay, rare Thai orchid species. Cryopreservation of protocorms was investigated by an encapsulation-dehydration method. The highest regrowth percentage of cryopreserved explants was observed when they were precultured on semi-solid ND medium with 0.25 M sucrose in the dark for 1 week then transferred to liquid ND medium containing 0.75 M sucrose for 2 days. The treated explants were dehydrated with silica gel in laminar air-flow for 5 h prior to immersion in liquid nitrogen for 1 h. Protocorm viability was tested by the TTC (2, 3, 5-triphenyltetrazoliumchloride) assay and regrowth ability was assessed by determining the survival percentage after recovery for 2 weeks. The survival rate of cryopreserved protocorms was 49% while percentage of TTC assay was 77%. Consequently, the simple and reliable protocol for cryopreservation of C. areitinum was clearly revealed in this study.

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