Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods

Two new cryopreservation techniques, PVS2 vitrification and encapsulationdehydration, were applied on PLBs of Dendrobium Bobby Messina, highly-prized orchid in Malaysia, with survival monitored through observations of growth and the 2,3,5-triphenyltetrazolium chloride (TTC) assay. In the vitrific...

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Main Author: Anthony, Jessica Jeyanthi James
Format: Thesis
Language:English
Published: 2012
Subjects:
Online Access:http://eprints.usm.my/45647/
http://eprints.usm.my/45647/1/Jessica%20James_HJ.pdf
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author Anthony, Jessica Jeyanthi James
author_facet Anthony, Jessica Jeyanthi James
author_sort Anthony, Jessica Jeyanthi James
building USM Institutional Repository
collection Online Access
description Two new cryopreservation techniques, PVS2 vitrification and encapsulationdehydration, were applied on PLBs of Dendrobium Bobby Messina, highly-prized orchid in Malaysia, with survival monitored through observations of growth and the 2,3,5-triphenyltetrazolium chloride (TTC) assay. In the vitrification method, the parameters optimized were PLBs size, preculture concentration, preculture durations and PVS2 incubation temperature and duration. In the encapsulation-dehydration method, the parameters optimized were PLBs size, preculture concentration, preculture duration and dehydration period with silica gel. In cryopreservation of Dendrobium Bobby Messina by vitrification technique, the optimized protocol based on TTC spectrophotometrical analysis and growth recovery were 3-4mm PLBs precultured in 0.2M sucrose for 1 day, treated with a mixture of 2M glycerol and 0.4M sucrose supplemented with half strength liquid MS media at 25°C for 20 minutes and subsequently dehydrated with plant vitrification solution 2 (PVS2) at 0°C for 20 minutes prior to storage in liquid nitrogen. Following rapid warming in a water bath at 40°C for 90 seconds, the PLBs were washed with half strength liquid MS media supplemented with 1.2M sucrose. Subsequently, the PLBs were cultured on half strength semi-solid MS media supplemented with 2% (w/v) sucrose without any growth regulators. The optimized vitrification method was successful in preserving this orchid as it produced growth recovery in non-cryopreservaed and cryopreserved PLBs up to 100% and 40% for Dendrobium Bobby Messina
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spelling usm-456472019-10-15T02:36:37Z http://eprints.usm.my/45647/ Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods Anthony, Jessica Jeyanthi James QH Natural history Two new cryopreservation techniques, PVS2 vitrification and encapsulationdehydration, were applied on PLBs of Dendrobium Bobby Messina, highly-prized orchid in Malaysia, with survival monitored through observations of growth and the 2,3,5-triphenyltetrazolium chloride (TTC) assay. In the vitrification method, the parameters optimized were PLBs size, preculture concentration, preculture durations and PVS2 incubation temperature and duration. In the encapsulation-dehydration method, the parameters optimized were PLBs size, preculture concentration, preculture duration and dehydration period with silica gel. In cryopreservation of Dendrobium Bobby Messina by vitrification technique, the optimized protocol based on TTC spectrophotometrical analysis and growth recovery were 3-4mm PLBs precultured in 0.2M sucrose for 1 day, treated with a mixture of 2M glycerol and 0.4M sucrose supplemented with half strength liquid MS media at 25°C for 20 minutes and subsequently dehydrated with plant vitrification solution 2 (PVS2) at 0°C for 20 minutes prior to storage in liquid nitrogen. Following rapid warming in a water bath at 40°C for 90 seconds, the PLBs were washed with half strength liquid MS media supplemented with 1.2M sucrose. Subsequently, the PLBs were cultured on half strength semi-solid MS media supplemented with 2% (w/v) sucrose without any growth regulators. The optimized vitrification method was successful in preserving this orchid as it produced growth recovery in non-cryopreservaed and cryopreserved PLBs up to 100% and 40% for Dendrobium Bobby Messina 2012-02 Thesis NonPeerReviewed application/pdf en http://eprints.usm.my/45647/1/Jessica%20James_HJ.pdf Anthony, Jessica Jeyanthi James (2012) Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods. Masters thesis, Universiti Sains Malaysia.
spellingShingle QH Natural history
Anthony, Jessica Jeyanthi James
Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods
title Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods
title_full Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods
title_fullStr Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods
title_full_unstemmed Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods
title_short Development Of Cryopreservation In Denrobium Bobby Messina Using PVS2 Vitrification And Encapsulation-Dehydration Methods
title_sort development of cryopreservation in denrobium bobby messina using pvs2 vitrification and encapsulation-dehydration methods
topic QH Natural history
url http://eprints.usm.my/45647/
http://eprints.usm.my/45647/1/Jessica%20James_HJ.pdf