CRYOPRESERVATION OF VANDA COERULEA PROTOCORM-LIKE BODIES BY DROPLET-VITRIFICATION

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 non-cryopreserved 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.

I International Symposium on Cryopreservation in Horticultural Species

N. Jitsopakul, K. Thammasiri, K. Ishikawa

cryopreservation, Vanda coerulea, protocorm-like bodies, droplet-vitrification, RAPD, cpDNA, trnL (UAA)

https://doi.org/10.17660/ActaHortic.2011.908.25