• cryopreservation of grapevine (vitis vinifera l.) in vitro shoot tips

    جزئیات بیشتر مقاله
    • تاریخ ارائه: 1392/07/24
    • تاریخ انتشار در تی پی بین: 1392/07/24
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     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 0°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).

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