Cryopreservation of protocorm-like bodies (PLBs) of Brassidium shooting star orchid by vitrification and encapsulation dehydration
2011
Hazirah Burkhan | Jessica Jeyanthi | Ranjetta Pobathy | Sreeramanan Subramaniam, Universiti Sains Malaysia, Minden Heights, 11800, Penang (Malaysia). School of Biological Sciences | Uma Rani Sinniah, Universiti Putra Malaysia, Serdang, 43400, Selangor (Malaysia). Dept. of Crop Sciences
Protocorm-like bodies (PLBs) of Brassidium Shooting Star, hybrid orchid which is a new commercial ornamental plant was cryopreserved by vitrification and encapsulation-dehydration techniques. The effects of PLB sizes, sucrose concentrations in preculture media and exposure to PVS2 duration were the main parameters assessed in vitrification technique. The viability of the cryopreserved PLBs were determined by 2,3,5-triphenyltetrazolium chloride (TTC) assay after two weeks post cryopreservation. In vitrification technique, PLBs with the size of 3-4 mm, precultured in half-strength semi-solid MS media supplemented with 0.8 M sucrose and dehydrated in PVS2 for 20 minutes at 0 deg C showed the best viability rate. In encapsulation dehydration technique, PLBs with the size of 3-4 mm precultured in half strength semi-solid MS media supplemented with 0.8 M sucrose and encapsulated in 3.5% of sodium alginate showed the best viability rate. The best parameters were chosen for positive and negative storage in LN and further biochemical content analysis (chlorophyll, total soluble protein and peroxidases activities) were conducted to investigate the physiological responses of the PLBs after cryopreservation.
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