The Development Of Encapsulationdehydration And Vitrification Protocols For Protocorm-Like Bodies (PLBs) Of Dendrobium sonia-28
| dc.contributor.author | Poobathy, Ranjetta | |
| dc.date.accessioned | 2018-07-18T06:26:12Z | |
| dc.date.available | 2018-07-18T06:26:12Z | |
| dc.date.issued | 2012-02 | |
| dc.description.abstract | The vitrification and encapsulation-dehydration methods of cryopreservation were applied on protocorm-like bodies (PLBs) of the orchid hybrid Dendrobium sonia-28, with survival assessments conducted through growth observations, visualand spectrophotometric 2,3,5-triphenyltetrazolium chloride (TTC) assays. The best survival (16.0% regeneration) for vitrification was obtained when 3-4mm PLBs were precultured in semi-solid half-strength MS medium supplemented with 0.4M sucrose, loaded in a loading solution for 20 minutes, dehydrated for 50 minutes at 0°C in plant vitrification solution 2 (PVS2), cryopreserved in liquid nitrogen (LN) for 24 hours, thawed in a 40±2°C water bath for 90 seconds, unloaded in 1.2M sucrose for 20 minutes, and regenerated in semi-solid half-strength MS medium containing 2g.L-1 charcoal, with all media supplemented with 0.6mM ascorbic acid. The PLBs were gradually exposed to light (darkness in the first week of recovery, exposure to 3.4μmol.m-2.s-1 for the subsequent three weeks and 150μmol.m-2.s-1 thereafter). The best viability rate (85.5% when observed for any signs of redness) for encapsulation-dehydration was obtained when 3-4mm PLBs were pretreated in semi-solid 0.5M sucrose medium, encapsulated for 30 minutes using 3% (w/v) liquid sodium alginate medium supplemented with 0.4M sucrose and 0.1M calcium chloride, osmoprotected in 0.75M sucrose solution for 24 hours at 25°C, dehydrated using 50g heat-sterilised silica gel for four hours, cryopreserved for 24 hours, thawed in a 40±2°C water bath for 90 seconds, and regenerated in semi-solid half-strengt MS medium followed by a transfer to medium supplemented with 1mg.L-1 6- benzyladenopurine (BAP), with gradual exposure to light (darkness in the first week of recovery, exposure to 3.4μmol.m-2.s-1 in the second week and 150μmol.m-2.s- 1thereafter). Biochemical analyses of PLBs subjected to the vitrificationcryopreservation experiment indicated a general decrease in the total protein content of cryopreserved PLBs with progression of the protocol. | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/5956 | |
| dc.language.iso | en | en_US |
| dc.publisher | Universiti Sains Malaysia | en_US |
| dc.subject | Encapsulation-dehydration and vitrification | en_US |
| dc.subject | protocols for protocorm-like bodies of dendrobium sonia-28 | en_US |
| dc.title | The Development Of Encapsulationdehydration And Vitrification Protocols For Protocorm-Like Bodies (PLBs) Of Dendrobium sonia-28 | en_US |
| dc.type | Thesis | en_US |
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