Cryopreservation of Vanda wightii Rchb.f protocorms
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Date
2014
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College of Agriculture, Vellayani
Abstract
Investigations on cryopreservation of protocorms of Vanda wightii Rchb.f an endangered orchid of Western Ghats was carried out to devise a protocol for long-term conservation of their germplasm. The commonly used techniques encapsulation- dehydration and vitrification methods were compared. Cryopreservation using encapsulation-dehydration gave maximum 36.1 per cent regeneration when one-step preculture was done in 0.5 M for 2 days and dehydrated for 4h. Optimum moisture content to get maximum recovery was 9.00 – 14%. Preculture in 0.5M sucrose for one day followed by 0.75M sucrose for one day did not improve regeneration of cryopreserved protocorms. Protocorms precultured in presence of 3 per cent DMSO did not recover after cryopreservation. Vitrification method tried as an alternative method gave 37.4 per cent regeneration when the protocorms were exposed to PVS2 for 2h prior to LN treatment. Exposure to PVS2 for more than 2h was detrimental to the protocorms so that Only 12.50 and 9.17 of the protocorms exposed for 3h and 4h period regenerated. Exposure to loading solution for 1h seems to give 34.95% regeneration after LN treatment suggesting that loading solution treatment alone is sufficient to cryoprotect the protocorms to get successful recovery after cryopreservation. Even though regeneration rates obtained through different methods are almost equal, consistent results and efficient direct plant regeneration make the vitrification method advantageous over encapsulation-dehydration method. Thus vitrification method is suggested as an efficient method for protocorm cryopreservation for germplasm conservation of V. wightii .
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