Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique

In vitro grown shoots of Brassia rex orchid hybrid was cryopreserved by means of plant vitrification solution 2 (PVS2) technique. For the preculture treatment, the shoots were excised into two standard sizes of 0.5-1.0 and 1.0-1.5 cm and were precultured on half-strength Murashige and Skoog (MS) sem...

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Main Authors: Johari, N., Chan, Lai Keng, Rathinam, Xavier, Sinniah, Uma Rani, Subramaniam, Sreeramanan
Format: Article
Language:English
English
Published: 2009
Online Access:http://psasir.upm.edu.my/id/eprint/12048/1/Cryopreservation%20of%20Brassia%20rex%20Orchid%20Shoots%20Using%20PVS2%20Technique.pdf
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author Johari, N.
Chan, Lai Keng
Rathinam, Xavier
Sinniah, Uma Rani
Subramaniam, Sreeramanan
author_facet Johari, N.
Chan, Lai Keng
Rathinam, Xavier
Sinniah, Uma Rani
Subramaniam, Sreeramanan
author_sort Johari, N.
collection UPM
description In vitro grown shoots of Brassia rex orchid hybrid was cryopreserved by means of plant vitrification solution 2 (PVS2) technique. For the preculture treatment, the shoots were excised into two standard sizes of 0.5-1.0 and 1.0-1.5 cm and were precultured on half-strength Murashige and Skoog (MS) semi solid medium supplemented with different concentrations of sucrose (control (0.06 M), 0.1, 0.25, 0.5 and 0.75 M) for 24 and 48 h. For the PVS2 dehydration treatment, the 0.1 M precultured (48 h and 1.0-1.5 cm) shoots were chosen for further experiment where the shoots were dehydrated in PVS2 solution at various durations (5, 10, 15, 20, 25 and 30 min) at 0 and 24°C for positive and negative storage in Liquid Nitrogen (LN). The viability of the cryopreserved cells were determined by 2, 3, 5-triphenyltetrazolium chloride (TTC) assay and chlorophyll extraction techniques. The best condition of PVS2 treatment was at 20 min of PVS2 treatment at 0°C prior to storage in liquid nitrogen. In chlorophyll determination based on chlorophyll assay, the highest concentration of total chlorophyll concentration (56.250 µg g-1) was obtained from shoots that were dehydrated for 25 min in PVS2 solution at 0°C without storage in liquid nitrogen.
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spelling upm.eprints-120482015-10-09T01:00:59Z http://psasir.upm.edu.my/id/eprint/12048/ Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique Johari, N. Chan, Lai Keng Rathinam, Xavier Sinniah, Uma Rani Subramaniam, Sreeramanan In vitro grown shoots of Brassia rex orchid hybrid was cryopreserved by means of plant vitrification solution 2 (PVS2) technique. For the preculture treatment, the shoots were excised into two standard sizes of 0.5-1.0 and 1.0-1.5 cm and were precultured on half-strength Murashige and Skoog (MS) semi solid medium supplemented with different concentrations of sucrose (control (0.06 M), 0.1, 0.25, 0.5 and 0.75 M) for 24 and 48 h. For the PVS2 dehydration treatment, the 0.1 M precultured (48 h and 1.0-1.5 cm) shoots were chosen for further experiment where the shoots were dehydrated in PVS2 solution at various durations (5, 10, 15, 20, 25 and 30 min) at 0 and 24°C for positive and negative storage in Liquid Nitrogen (LN). The viability of the cryopreserved cells were determined by 2, 3, 5-triphenyltetrazolium chloride (TTC) assay and chlorophyll extraction techniques. The best condition of PVS2 treatment was at 20 min of PVS2 treatment at 0°C prior to storage in liquid nitrogen. In chlorophyll determination based on chlorophyll assay, the highest concentration of total chlorophyll concentration (56.250 µg g-1) was obtained from shoots that were dehydrated for 25 min in PVS2 solution at 0°C without storage in liquid nitrogen. 2009 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/12048/1/Cryopreservation%20of%20Brassia%20rex%20Orchid%20Shoots%20Using%20PVS2%20Technique.pdf Johari, N. and Chan, Lai Keng and Rathinam, Xavier and Sinniah, Uma Rani and Subramaniam, Sreeramanan (2009) Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique. Research Journal of Botany, 4 (3). pp. 74-88. ISSN 1816-4919 10.3923/rjb.2009.74.88 English
spellingShingle Johari, N.
Chan, Lai Keng
Rathinam, Xavier
Sinniah, Uma Rani
Subramaniam, Sreeramanan
Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique
title Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique
title_full Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique
title_fullStr Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique
title_full_unstemmed Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique
title_short Cryopreservation of Brassia rex Orchid Shoots Using PVS2 Technique
title_sort cryopreservation of brassia rex orchid shoots using pvs2 technique
url http://psasir.upm.edu.my/id/eprint/12048/1/Cryopreservation%20of%20Brassia%20rex%20Orchid%20Shoots%20Using%20PVS2%20Technique.pdf
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AT sinniahumarani cryopreservationofbrassiarexorchidshootsusingpvs2technique
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