Articles
CRYOPRESERVATION BY ENCAPSULATION-DEHYDRATION FOR LONG-TERM STORAGE OF AERIDES FALCATE LINDL.
Article number
1039_19
Pages
155 – 160
Language
English
Abstract
Khok Phu Ta Ka, a plant sanctuary in Khonkaen province, is one of Thailands important natural resources of wild orchids.
Today a number of wild orchid species are being threatened and are in danger of extinction due to habitat destruction and commercial exploitation such as plant collection for local trade and deforestation for the expansion of agriculture.
As cryopreservation is considered efficient for long-term plant germplasm storage, it has therefore been employed for germplasm conservation in many plant species.
The main purpose of the present study was to develop an efficient protocol for long-term storage of Aerides falcate Lindl protocorms.
To establish the efficient protocol, protocorms were encapsulated in Ca-alginate, followed by pretreatments with sucrose.
Prior to immersion into liquid nitrogen, pretreated protocorms were dehydrated by air-drying.
The efficient protocol for cryopreservation was achieved by the pretreatment of encapsulated protocorms with 0.5 M sucrose for 18 h at 25°C, followed by 6 h of dehydration before plunging encapsulated protocorms into liquid nitrogen.
The percentage of viable protocorms was 82. Flow cytometry showed no difference of DNA content between treated groups and control groups.
In summary, the protocol obtained from the present study is simple, non-laborious and effective.
Today a number of wild orchid species are being threatened and are in danger of extinction due to habitat destruction and commercial exploitation such as plant collection for local trade and deforestation for the expansion of agriculture.
As cryopreservation is considered efficient for long-term plant germplasm storage, it has therefore been employed for germplasm conservation in many plant species.
The main purpose of the present study was to develop an efficient protocol for long-term storage of Aerides falcate Lindl protocorms.
To establish the efficient protocol, protocorms were encapsulated in Ca-alginate, followed by pretreatments with sucrose.
Prior to immersion into liquid nitrogen, pretreated protocorms were dehydrated by air-drying.
The efficient protocol for cryopreservation was achieved by the pretreatment of encapsulated protocorms with 0.5 M sucrose for 18 h at 25°C, followed by 6 h of dehydration before plunging encapsulated protocorms into liquid nitrogen.
The percentage of viable protocorms was 82. Flow cytometry showed no difference of DNA content between treated groups and control groups.
In summary, the protocol obtained from the present study is simple, non-laborious and effective.
Authors
S. Bunnag, J. Hongthongkham
Keywords
Khok Phu Ta Ka, liquid nitrogen, naphthaleneacetic acid, 6-benzylaminopurine, flow cytometry
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