Cryopreservation of in vitro sugar beet (Beta vulgaris L.) shoot tips by a vitrification technique |
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Authors: | B Vandenbussche G Weyens M De Proft |
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Institution: | (1) Laboratory of Plant Culture, Catholic University of Leuven, Willem de Croylaan 42, 3001 Heverlee, Belgium e-mail: bert.vandenbussche@agr.kuleuven.ac.be Fax: +32-16-322966, BE;(2) ADVANTA, Industriepark Soldatenplein Z2 15, 3300 Tienen, Belgium, BE |
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Abstract: | Sugar beet shoot tips from cold-acclimated plants were successfully cryopreserved using a vitrification technique. Dissected
shoot tips were precultured for 1 day at 5 °C on solidified DGJ0 medium with 0.3 M sucrose. After loading for 20 min with
a mixture of 2 M glycerol and 0.4 M sucrose (20 °C), shoot tips were dehydrated with PVS2 (0 °C) for 20 min prior to immersion
in liquid nitrogen. Both cold acclimation and loading enhanced the dehydration tolerance of shoot tips to PVS2. After thawing,
shoot tips were deloaded for 15 min in liquid DGJ0 medium with 1.2 M sucrose (20 °C). The optimal exposure time to both loading
solution and PVS2 depended on the in vitro morphology of the clone. With tetraploid clones a higher sucrose concentration
during cold acclimation and preculture further enhanced survival after cryopreservation. Survival rates ranged between 60%
and 100% depending on the clone. Since only 10–50% of the surviving shoot tips developed into non-hyperhydric shoots, regrowth
was optimized.
Received: 13 September 1999 / Revision received: 2 March 2000 / Accepted: 16 March 2000 |
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Keywords: | Beta vulgaris L Cryopreservation Shoot tips Encapsulation-dehydration Vitrification |
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