Location: Plant Germplasm Preservation ResearchTitle: Successful cryopreservation of Vitis vinifera cv. ‘Chardonnay’ from both in vitro and growth chamber source plants Author
|Bettoni, Jean - University Of Santa Catarina|
|Kretzschmar, Aike Anneliese - University Of Santa Catarina|
Submitted to: Acta Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/24/2018
Publication Date: 2/28/2019
Citation: Bettoni, J.C., Bonnart, R.M., Shepherd, A.N., Kretzschmar, A., Volk, G.M. 2019. Successful cryopreservation of Vitis vinifera cv. ‘Chardonnay’ from both in vitro and growth chamber source plants. Acta Horticulturae. 1234:211-218. https://doi.org/10.17660/ActaHortic.2019.1234.28.
DOI: https://doi.org/10.17660/ActaHortic.2019.1234.28 Interpretive Summary: Genebank collections of grapes (Vitis) are usually maintained in field conditions. As such, they are vulnerable to abiotic and biotic threats. We sought to identify strategies whereby field collections of Vitis could be securely backed-up as cryopreserved propagules at the USDA-ARS National Laboratory for Genetic Resources Preservation in Fort Collins, Colorado. We identified a cryopreservation method for the Vitis vinifera cultivar 'Chardonnay' by which shoot tips excised from either in vitro or growth chamber source plants could be successfully cryopreserved. The high levels of viability after liquid nitrogen exposure using both in vitro (65% regrowth levels) and growth chamber (43% regrowth levels) stock plants suggest that it may be possible to cryopreserve Vitis shoot tips without first introducing each accession into tissue culture. The use of growth chamber source plants would improve the efficiency of the cryopreservation process when it is implemented.
Technical 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 at a secondary site. 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 22oC, prior to full-strength PVS2 treatment at 0oC. Shoot tips derived from in vitro grown stock plants had the highest regrowth level of 65% with a PVS2 exposure duration of 75 min at 0oC. 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 0oC. 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.