Articles
Successful cryopreservation of Vitis vinifera ‘Chardonnay’ from both in vitro and growth chamber source plants
Article number
1234_28
Pages
211 – 218
Language
English
Abstract
Both the United States and Brazil maintain genebank collections of vegetatively propagated crops, including grapevine (Vitis). Vitis collections are expensive to maintain in the field and are vulnerable to abiotic and biotic threats.
When robust methods are available, cryopreservation provides an opportunity to securely back-up collections with lower maintenance costs than those of duplicated field collections.
We have developed a droplet-vitrification method that results in high levels of shoot tip regrowth of Vitis vinifera cultivar ‘Chardonnay’ after liquid nitrogen exposure.
Uniform shoot tips were obtained from nodal sections cultured from in vitro or growth chamber stock plants.
Pretreatments included 0.3 M sucrose, salicylic acid, ascorbic acid, and glutathione (reduced form). Half-strength PVS2 was applied for 30 minutes at 22°C, prior to full-strength PVS2 treatment at 0°C. Shoot tips derived from in vitro grown stock plants had the highest regrowth level of 68% with a PVS2 exposure duration of 75 min at 0°C. Shoot tips derived from growth chamber grown stock plants had the highest regrowth level of 43% with a PVS2 exposure duration of 30 min at 0°C. The high levels of regrowth using both in vitro and growth chamber stock plants suggest that it may be possible to cryopreserve Vitis shoot tips without first introducing each accession into tissue culture.
When robust methods are available, cryopreservation provides an opportunity to securely back-up collections with lower maintenance costs than those of duplicated field collections.
We have developed a droplet-vitrification method that results in high levels of shoot tip regrowth of Vitis vinifera cultivar ‘Chardonnay’ after liquid nitrogen exposure.
Uniform shoot tips were obtained from nodal sections cultured from in vitro or growth chamber stock plants.
Pretreatments included 0.3 M sucrose, salicylic acid, ascorbic acid, and glutathione (reduced form). Half-strength PVS2 was applied for 30 minutes at 22°C, prior to full-strength PVS2 treatment at 0°C. Shoot tips derived from in vitro grown stock plants had the highest regrowth level of 68% with a PVS2 exposure duration of 75 min at 0°C. Shoot tips derived from growth chamber grown stock plants had the highest regrowth level of 43% with a PVS2 exposure duration of 30 min at 0°C. The high levels of regrowth using both in vitro and growth chamber stock plants suggest that it may be possible to cryopreserve Vitis shoot tips without first introducing each accession into tissue culture.
Authors
J.C. Bettoni, R. Bonnart, A. Shepherd, A.A. Kretzschmar, G.M. Volk
Keywords
cryopreservation, droplet-vitrification, genebanking, grapevine
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