Cryopreservation of in vitro-grown shoot tips of grapevine by encapsulation-dehydration |
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Authors: | Wang Qiaochun Tanne Edna Arav Amir Gafny Ron |
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Institution: | (1) Department of Virology, Israel;(2) Institute of Animal Science, Agricultural Research Organization, The Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel |
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Abstract: | In vitro-grown shoot tips of the LN33 hybrid (Vitis L.) and cv. Superior (Vitis vinifera L.) were successfully cryopreserved by encapsulation-dehydration. Encapsulated shoot tips were precultured stepwise on half-strength
MS medium supplemented with increasing sucrose concentrations of 0.25, 0.5, 0.75 and 1.0 M for 4 days, with one day for each
step. Following preculture, encapsulated shoot tips were dehydrated prior to direct immersion in liquid nitrogen for 1 h.
After thawing, cryopreserved shoot tips were post-cultured on a post-culture medium for survival. An optimal survival of cryopreserved
shoot tips was achieved when encapsulated shoot tips were dehydrated to 15.6 and 17.6% water content for the LN33 hybrid and
cv. Superior, respectively. Comparison between the effects of dehydration with silica gel and by air drying on cryopreserved
shoot tips, showed that survival was dependent on water content, not on dehydration method. The thawing method markedly affected
survival of cryopreserved shoot tips, and thawing at 40 °C for 3 min was found best. No callus formation and fastest shoot
elongation were obtained when cryopreserved shoot tips were post-cultured on the post-culture medium composed of half-strength
MS supplemented with 1 mg l−1 BA and 0.1 mg l−1 NAA. With these optimized parameters, 60 and 40% survival of cryopreserved shoot tips were obtained for the LN33 hybrid and
cv. Superior, respectively.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | in vitro culture long-term storage shoot apex Vitis spp woody plant |
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