Articles
CRYOPRESERVATION BY VITRIFICATION OF EMBRYOGENIC CALLUS OF WILD CROCUS (CROCUS HYEMALIS AND CROCUS MOABITICUS)
Article number
908_30
Pages
239 – 246
Language
English
Abstract
Cryopreservation of embryogenic callus of the wild crocus species (Crocus hyemalis and Crocus moabiticus) was studied using vitrification-based techniques.
The effect of PVS2 concentration on the viability of crocus calli before and after cryopreservation was examined.
Higher viability of calli was obtained using step-wise vitrification than direct exposure to a 100% PVS2. Complete survival and 91.2-94.3% regrowth rates of C. hyemalis and C. moabiticus calli, respectively, were achieved with four-step dehydration by PVS2 for 20 min prior to freezing.
The effect of different cryoprotectants on survival of crocus calli was also tested.
The use of 2 M glycerol plus 0.4 M sucrose resulted in 92.2% survival of C. hyemalis calli and 97.4% survival of C. moabiticus prior to freezing.
Furthermore, the effect of combinations of different cryoprotectants and vitrification solutions on viability of calli was tested.
Cryoprotection by 2 M glycerol plus 0.4 M sucrose followed by dehydration with 15% DMSO plus 1.0 M sucrose vitrification solution gave complete survival and maximum regrowth (21.1-25.0%) rates after freezing for C. hyemalis and C. moabiticus calli, respectively.
The effect of PVS2 concentration on the viability of crocus calli before and after cryopreservation was examined.
Higher viability of calli was obtained using step-wise vitrification than direct exposure to a 100% PVS2. Complete survival and 91.2-94.3% regrowth rates of C. hyemalis and C. moabiticus calli, respectively, were achieved with four-step dehydration by PVS2 for 20 min prior to freezing.
The effect of different cryoprotectants on survival of crocus calli was also tested.
The use of 2 M glycerol plus 0.4 M sucrose resulted in 92.2% survival of C. hyemalis calli and 97.4% survival of C. moabiticus prior to freezing.
Furthermore, the effect of combinations of different cryoprotectants and vitrification solutions on viability of calli was tested.
Cryoprotection by 2 M glycerol plus 0.4 M sucrose followed by dehydration with 15% DMSO plus 1.0 M sucrose vitrification solution gave complete survival and maximum regrowth (21.1-25.0%) rates after freezing for C. hyemalis and C. moabiticus calli, respectively.
Authors
S.H. Baghdadi, I. Makhadmeh, M. Syouf, A. Arabiat, R.A. Shibli, M.A. Shatnawai
Keywords
Crocus, DMSO, TTC, PVS2, vitrification
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