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|Title:||Cryopreservation of Vanda coerulea protocorm-like bodies by droplet-vitrification|
|Keywords:||cpDNA;Cryopreservation;Dropletvitrification;Protocorm-like bodies;RAPD;trnL (UAA);Vanda coerulea|
|Publisher:||Copyright 2012 Elsevier B.V.|
|Citation:||Acta Horticulturae 908 (2011);207-214|
|Abstract:||Vanda coerulea is the most popular Vanda species of Thailand that has become endangered because of the wild orchid trade, deforestation and environmental changes. To preserve this orchid, droplet-vitrification was studied for the cryopreservation of protocorm-like bodies. Protocorm-like bodies were developed from one protocrom in ND liquid medium supplemented with 1 mg/L BA in combination with 0.5 mg/L NAA and 30 g/L maltose. Protocorm-like bodies (3.0 mm in diameter) were precultured in ND liquid medium supplemented with 0.5 M sucrose for 1 day on a shaker, then dehydrated with loading solution for 15 min and exposed to PVS2 solution for 30 min at 25°C. The plant materials were then immersed in liquid nitrogen, rewarmed rapidly and cultured on solidified ND medium. After cryopreservation, the survival frequency of cryopreserved protocorm-like bodies was 5%. The assessment of the genetic stability of cryopreserved protocorm-like bodies was developed using RAPD markers, size and sequence of the trnL (UAA) non-coding region of cpDNA. Results showed the same RAPD patterns of non-cryopreserved and cryopreserved protocorm-like bodies. The size and sequences of the trnL (UAA) non-coding region of cpDNA for noncryopreserved and cryopreserved protocorm-like bodies were not different. There was no difference in morphology and similar patterns of ploidy in plantlets developed from non-cryopreserved and cryopreserved protocorm-like bodies.|
|Appears in Collections:||Plant Science: International Proceedings|
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