Search results for: Sutha Klaocheed

2 Enhanced Efficiency for Propagation of Phalaenopsis cornu-cervi (Breda) Blume & Rchb. F. Using Trimmed Leaf Technique

Authors: Suphat Rittirat, Sutha Klaocheed, Kanchit Thammasiri

Abstract:

The effects of thidiazuron (TDZ) and benzyladenine (BA) on protocorm-like bodies (PLBs) induction from leaf explants was investigated. It was found that TDZ was superior to BA. The highest percentage and number of PLBs per leaf explant at 30 and 5.3 respectively were obtained on ½ MS medium supplemented with 9µM TDZ. The regenerated plantlets were potted and acclimatized in the greenhouse. These plants grew well and developed into normal plants after 3 month of transplantation. The 100% survival of plantlets was achieved when planted on pots containing sphagnum moss.

Keywords: orchid, PLBs, sphagnum moss, thidiazuron

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1 Vitrification-Based Cryopreservation of Phalaenopsis cornu-Cervi (Breda) Blume & Rchb. f. Protocorms

Authors: Suphat Rittirat, Sutha Klaocheed, Somporn Prasertsongskun, Kanchit Thammasiri

Abstract:

Protocorms of Phalaenopsis cornu-cervi (Breda) Blume & Rchb. f. were successfully cryopreserved using a vitrification method. Two-month old protocorms at GI 4 stage were precultured in liquid MS medium supplemented with different concentrations of sucrose (0.3, 0.5, 0.7, 0.9 and 1.2 M) at 25±1°C for 2 days on an orbital shaker at 110 rpm. The protocorms were treated with loading solution (2 M glycerol plus 0.4 M sucrose) for 20 minutes at 25±1°C. Then, the protocorms were sufficiently dehydrated with vitrification solution (plant vitrification solution 2, PVS2) for various times (0, 30, 60, 90 and 120 minutes) at 25±1°C and stored in liquid nitrogen for 1 day. After rapid thawing in water bath at 40°C for 2 minutes, the explants were washed by MS liquid medium containing 0.5 ml of 1.2 M sucrose for 20 minutes. The results shown that the protocorms were precultured in liquid MS medium containing 0.5 M sucrose and dehydrated with vitrification solution for 60 minutes had the highest survival percentage of protocorm at 31±1.0 % as measured by Evan’s blue. No survival rate of protocorms was found without vitrification treatments.

Keywords: protocorms, cryopreservation, Phalaenopsis cornu-cervi, vitrification

Procedia PDF Downloads 336