Cryopreservation Of Brassidium Shooting Star Orchid Using Dropletvitrification Method

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dc.date.accessioned2017-11-07T06:48:45Z
dc.date.available2017-11-07T06:48:45Z
dc.date.issued2014-07
dc.description.abstractThe aims of this research are to investigate the efficiency of Protocorm-like bodies (PLBs) induction method and develop a droplet-vitrification protocol applicable for cryopreservation of Brassidium Shooting Star orchid. The effect of plant growth regulators such as benzylaminopurine (BAP), naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D) on PLBs formation of Brassidium Shooting Star orchid was studied. Basal corms were cultured in media supplemented with BA alone or in combination with NAA and 2,4-D. The numbers of PLBs induced from each explant were counted after six weeks of treatment. Addition of 1.0 mg/L BAP to MS basal media was found to be the best media for PLBs induction that producing an average of 25.43±5.12 PLB per explant. Seven (7) parameters have been optimised in droplet-vitrification method such as PLB size, sugar type, preculture concentration, preculture incubation duration, plant vitrification solution 2 (PVS2) incubtion duration, thawing method and unloading durations. Assessments of viability were evaluated by using TTC spectrophotometric at 490nm absorbance reading and growth recovery the regenerated PLBs. The highest growth recovery of 30% was obtained with 3-4 mm PLBs precultured on media supplemented with 0.25M sucrose for 7 days, followed by loading treatment for 20 min, dehydration with PVS2 solution for 40 min and treated with unloading solution for 15 min. In order to reveal the lethal and non-lethal damages produced by cryopreservation, histological, scanning electron microscopy and biochemical analysis were carried out on both cryopreserved and non-cryopreserved PLBs of Brassidium Shooting Star orchid comparing with the PLBs stock culture. Histological analysis displayed cryopreservation protocol caused structural changes in cryopreserved PLBs and non-cryopreserved PLB. Similarly scanning electron micrograph also showed severe damages in cryopreserved and non-cryopreserved PLBs comparative to the PLBs stock culture. Addition of ascorbic acid and α- tocopherol which reported to improve regrowth of cryopreserved plant did not affect the regrowth of Brassidium Shooting Star orchid PLBs. However, addition α- tocopherol promoted faster recovery of cryopreserved PLBs.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/5242
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectEfficiency of Protocorm-like bodiesen_US
dc.subjectcryopreservation of Brassidium Shooting Star orchiden_US
dc.titleCryopreservation Of Brassidium Shooting Star Orchid Using Dropletvitrification Methoden_US
dc.typeThesisen_US
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