Markovic Z., Chatelet P., Sylvestre Isabelle, Kontic J. K., Engelmann Florent. (2013). Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips. Central European Journal of Biology, 8 (10), p. 993-1000. ISSN 1895-104X.
Titre du document
Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips
Markovic Z., Chatelet P., Sylvestre Isabelle, Kontic J. K., Engelmann Florent
Source
Central European Journal of Biology, 2013,
8 (10), p. 993-1000 ISSN 1895-104X
In this work, we compared the efficiency of encapsulation-dehydration and droplet-vitrification techniques for cryopreserving grapevine (Vitis vinifera L.) cv. Portan shoot tips. Recovery of cryopreserved samples was achieved with both techniques; however, droplet-vitrification, which was used for the first time with grapevine shoot tips, produced higher regrowth. With encapsulationdehydration, encapsulated shoot tips were precultured in liquid medium with progressively increasing sucrose concentrations over a 2-day period (12 h in medium with 0.25, 0.5, 0.75 and 1.0 M sucrose), then dehydrated to 22.28% moisture content (fresh weight). After liquid nitrogen exposure 37.1% regrowth was achieved using 1 mm-long shoot tips and only 16.0% with 2 mm-long shoot tips. With droplet-vitrification, 50% regrowth was obtained following treatment of shoot tips with a loading solution containing 2 M glycerol + 0.4 M sucrose for 20 min, dehydration with half-strength PVS2 vitrification solution (30% (w/v) glycerol, 15% (w/v) ethylene glycol, 15% dimethylsulfoxide and 0.4 M sucrose in basal medium) at room temperature, then with full strength PVS2 solution at 0A degrees C for 50 min before direct immersion in liquid nitrogen. No regrowth was achieved after cryopreservation when shoot tips were dehydrated with PVS3 vitrification solution (50% (w/v) glycerol and 50% (w/v) sucrose in basal medium).
Plan de classement
Sciences du monde végétal [076]
;
Biotechnologies [084]
