Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid

Protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid were successfully cryopreserved using droplet-vitrification method. Vitrification based cryopreservation protocol is comprised of preculture, osmoprotection, cryoprotection, cooling, rewarming, and growth recovery and each and every s...

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Main Authors: Rahmah, Safrina, Ahmad Mubbarakh, Safiah, Khor, Soo Ping, Subramaniam, Sreeramanan
Format: Article
Language:English
Published: Hindawi Publishing Corporation 2015
Subjects:
Online Access:http://eprints.usm.my/38896/
http://eprints.usm.my/38896/1/Effects_of_Droplet-Vitrification_Cryopreservation.pdf
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author Rahmah, Safrina
Ahmad Mubbarakh, Safiah
Khor, Soo Ping
Subramaniam, Sreeramanan
author_facet Rahmah, Safrina
Ahmad Mubbarakh, Safiah
Khor, Soo Ping
Subramaniam, Sreeramanan
author_sort Rahmah, Safrina
building USM Institutional Repository
collection Online Access
description Protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid were successfully cryopreserved using droplet-vitrification method. Vitrification based cryopreservation protocol is comprised of preculture, osmoprotection, cryoprotection, cooling, rewarming, and growth recovery and each and every step contributes to the achievement of successful cryopreservation. In order to reveal the lethal and nonlethal damage produced by cryopreservation, histological observation, scanning electron microscopy (SEM), and biochemical analysis were carried out in both cryopreserved and noncryopreserved PLBs of Brassidium Shooting Star orchid comparing with the control PLBs stock culture. Histological and scanning electron microscopy analyses displayed structural changes in cryopreserved PLBs due to the impact of cryoinjury during exposure to liquid nitrogen. Total soluble protein significantly increased throughout the dehydration process and the highest value was achieved when PLBs were stored in liquid nitrogen. Ascorbate peroxidase (APX) and catalase (CAT) showed the highest enzyme activities in both dehydration and cryostorage treatments indicating that stress level of PLBs was high during these stages.
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spelling usm-388962018-02-14T00:28:13Z http://eprints.usm.my/38896/ Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid Rahmah, Safrina Ahmad Mubbarakh, Safiah Khor, Soo Ping Subramaniam, Sreeramanan QH1 Natural history (General - Including nature conservation, geographical distribution) Protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid were successfully cryopreserved using droplet-vitrification method. Vitrification based cryopreservation protocol is comprised of preculture, osmoprotection, cryoprotection, cooling, rewarming, and growth recovery and each and every step contributes to the achievement of successful cryopreservation. In order to reveal the lethal and nonlethal damage produced by cryopreservation, histological observation, scanning electron microscopy (SEM), and biochemical analysis were carried out in both cryopreserved and noncryopreserved PLBs of Brassidium Shooting Star orchid comparing with the control PLBs stock culture. Histological and scanning electron microscopy analyses displayed structural changes in cryopreserved PLBs due to the impact of cryoinjury during exposure to liquid nitrogen. Total soluble protein significantly increased throughout the dehydration process and the highest value was achieved when PLBs were stored in liquid nitrogen. Ascorbate peroxidase (APX) and catalase (CAT) showed the highest enzyme activities in both dehydration and cryostorage treatments indicating that stress level of PLBs was high during these stages. Hindawi Publishing Corporation 2015 Article PeerReviewed application/pdf en http://eprints.usm.my/38896/1/Effects_of_Droplet-Vitrification_Cryopreservation.pdf Rahmah, Safrina and Ahmad Mubbarakh, Safiah and Khor, Soo Ping and Subramaniam, Sreeramanan (2015) Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid. Scientific World Journal, 2015 (961793). pp. 1-10. ISSN 2356-6140 http://dx.doi.org/10.1155/2015/961793
spellingShingle QH1 Natural history (General - Including nature conservation, geographical distribution)
Rahmah, Safrina
Ahmad Mubbarakh, Safiah
Khor, Soo Ping
Subramaniam, Sreeramanan
Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid
title Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid
title_full Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid
title_fullStr Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid
title_full_unstemmed Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid
title_short Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid
title_sort effects of droplet-vitrification cryopreservation based on physiological and antioxidant enzyme activities of brassidium shooting star orchid
topic QH1 Natural history (General - Including nature conservation, geographical distribution)
url http://eprints.usm.my/38896/
http://eprints.usm.my/38896/
http://eprints.usm.my/38896/1/Effects_of_Droplet-Vitrification_Cryopreservation.pdf