Naslov
Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips using encapsulation-dehydration and droplet-vitrification

Autori
Marković, Zvjezdana ; Chatelet, Philippe ; Sylvestre, Isabelle ; Karoglan Kontić, Jasminka ; Engelmann, Florent

Izvornik
Central european journal of biology (1895-104X) 8 (2013), 10; 993-1000

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Vitis vinifera L.; cryopreservation; encapsulation-dehydration; droplet-vitrification.

Sažetak
In this work, we compared the efficiency of the encapsulation-dehydration and droplet-vitrification techniques for cryopreserving shoot tips of grapevine (Vitis vinifera L.) cv. Portan. 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 encapsulation-dehydration, encapsulated shoot tips were precultured in liquid medium with progressively increasing sucrose concentrations over a period of 2 days (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 0°C for 50 min before direct immersion in liquid nitrogen. No regrowth was achieved after cryopreservation when shoot tips were dehydrated with the PVS3 vitrification solution (50% (w/v) glycerol and 50% (w/v) sucrose in basal medium).

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Poljoprivreda (agronomija), Biotehnologija

Napomena
Rad je nastao u istraživačkom projektu uz potporu stipendije Francuske vlade u Hrvatskoj.

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