Articles
Cryopreservation of protocorm-like bodies (PLBs) of Phalaenopsis cornu-cervi (Breda) Blume & Rchb. f. by vitrification approach
Article number
1421_13
Pages
95 – 102
Language
English
Abstract
Cryopreservation can be a stable, long-term method of germplasm conservation, but successful application can be challenging for tropical material.
Cryopreservation provides a means of storing biological material at ultra-low temperatures, normally that of liquid nitrogen, LN, (-196°C). Protocorm-like bodies (PLBs) of Phalaenopsis cornu-cervi (Breda) Blume & Rchb. f. were successfully cryopreserved by vitrification approach.
The sucrose concentrations in preculture media and exposure to PVS2 duration were the main parameters assessed in vitrification approach.
In vitrification approach, PLBs with the size of 3-4 mm, precultured in MS liquid medium supplemented with 0.5 M sucrose at 25±2°C for 2 d on an orbital shaker at 110 rpm., treated with loading solution (2 M glycerol plus 0.4 M sucrose) for 10 min at 25±2°C followed by dehydration with plant vitrification solution 2 (PVS2) for 60 min at 0°C prior to cryopreservation had the highest survival rate of percentage of cryopreserved PLBs at 31±1.0% as measured by Evan’s blue.
No survival rate of cryopreserved PLBs was found without vitrification treatments.
Establishment of this protocol provides an alternative means for cryopreservation of Phalaenopsis.
Cryopreservation provides a means of storing biological material at ultra-low temperatures, normally that of liquid nitrogen, LN, (-196°C). Protocorm-like bodies (PLBs) of Phalaenopsis cornu-cervi (Breda) Blume & Rchb. f. were successfully cryopreserved by vitrification approach.
The sucrose concentrations in preculture media and exposure to PVS2 duration were the main parameters assessed in vitrification approach.
In vitrification approach, PLBs with the size of 3-4 mm, precultured in MS liquid medium supplemented with 0.5 M sucrose at 25±2°C for 2 d on an orbital shaker at 110 rpm., treated with loading solution (2 M glycerol plus 0.4 M sucrose) for 10 min at 25±2°C followed by dehydration with plant vitrification solution 2 (PVS2) for 60 min at 0°C prior to cryopreservation had the highest survival rate of percentage of cryopreserved PLBs at 31±1.0% as measured by Evan’s blue.
No survival rate of cryopreserved PLBs was found without vitrification treatments.
Establishment of this protocol provides an alternative means for cryopreservation of Phalaenopsis.
Authors
S. Rittirat, S. Klaocheed, S. Kalawong, K. Thammasiri
Keywords
epiphytes, ex situ conservation, germplasm conservation, liquid nitrogen, Orchidaceae
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