Articles
TWO-STEP PRECONDITIONING – A FEASIBLE METHOD FOR CRYOPRESERVATION OF FORTUNELLA POLYANDRA SHOOT TIPS USING VITRIFICATION TECHNIQUE
Article number
908_38
Pages
289 – 296
Language
English
Abstract
A study was conducted to develop an optimized methodology for cryopreservation of in vitro derived Fortunella polyandra shoot tips using vitrification-based techniques.
Cryopreservation of F. polyandra shoot tips was successfully performed with the emphasis placed upon the aspects of preconditioning involving the pregrowth and preculture of explants.
Preculture alone on Murashige and Skoog (MS) medium with 0.3 M sucrose for one day was not sufficient in giving survival after cryopreservation.
A preliminary experiment involving pregrowth on MS medium with 0.72 M sucrose for three days produced a survival of 23.4% after cryopreservation.
The usage of a combination of cryoprotectants in the preculture medium (MS with 0.3 M sucrose and 0.5 M ethylene glycol) proved to be beneficial in enhancing survival percentage of cryopreserved explants up to 40% when precultured for one day.
However the highest percentage of shoot tip survival after cryopreservation (56.6%) was achieved when a two-step preconditioning method (pregrowth on MS medium with 0.72 M sucrose for three days followed by preculture on MS medium with 0.3 M sucrose for one day) was employed.
Cryopreservation of F. polyandra shoot tips was successfully performed with the emphasis placed upon the aspects of preconditioning involving the pregrowth and preculture of explants.
Preculture alone on Murashige and Skoog (MS) medium with 0.3 M sucrose for one day was not sufficient in giving survival after cryopreservation.
A preliminary experiment involving pregrowth on MS medium with 0.72 M sucrose for three days produced a survival of 23.4% after cryopreservation.
The usage of a combination of cryoprotectants in the preculture medium (MS with 0.3 M sucrose and 0.5 M ethylene glycol) proved to be beneficial in enhancing survival percentage of cryopreserved explants up to 40% when precultured for one day.
However the highest percentage of shoot tip survival after cryopreservation (56.6%) was achieved when a two-step preconditioning method (pregrowth on MS medium with 0.72 M sucrose for three days followed by preculture on MS medium with 0.3 M sucrose for one day) was employed.
Authors
D.C. Chandrabalan, M.M. Clyde, M.N. Normah
Keywords
pregrowth, preculture, sucrose, ethylene glycol, cryoprotectant
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