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Title: The development of a droplet-vitrification method to conserve Vitis collections in the USDA-ARS National Plant Germplasm System and UDESC-CAV Santa Catarina State University in Brazil

Author
item BETTONI, JEAN - University Of Santa Catarina
item Bonnart, Remi
item Shepherd, Ashley
item KRETZSCHMAR, AIKE - Universidade Federal De Santa Catarina (UFSC)
item Volk, Gayle

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 1/11/2018
Publication Date: 3/26/2018
Citation: Bettoni JC, Bonnart R, Shepherd A, Kretzschmar AA, Volk GM. 2018. The development of a droplet-vitrification method to conserve Vitis collections in the USDA-ARS National Plant Germplasm System and UDESC-CAV Santa Catarina State University in Brazil. Third International Symposium on Plant Cryopreservation. CryoSymp 2018. 26-28 March 2018. Bangkok, Thailand. pg. 24.

Interpretive Summary: Both the United States and Brazil maintain vast collections of grape genetic resources. We share a common interest in using cryopreservation methods for the secure, long-term back-up of accessions within these field collections of the USDA-ARS National Plant Germplasm System and UDESC-CAV Santa Catarina State University in Brazil. We have a developed a Vitis droplet-vitrification method that results in high levels of shoot tip regrowth after liquid nitrogen exposure. Uniform shoot tips were obtained from nodal sections of either in vitro or growth chamber stock plants. Pretreatments included 0.3 M sucrose, salicylic acid, ascorbic acid, and glutathione. Half-strength PVS2 was applied for 30 minutes at 25oC, prior to full-strength PVS2 treatment at 0oC. The optimum PVS2 exposure duration was 30 minutes for growth chamber-derived shoot tips and 90 minutes for in vitro derived shoot tips. Shoot tips were then placed onto foil strips and plunged into LN. Regrowth levels ranged from 45 to 72 % for the seven Vitis species tested from in vitro plants, and from 35 to 65 % for three Vitis vinifera cultivars and one Vitis hybrid tested from growth chamber plants. The high levels of regrowth suggest that Vitis cryopreservation may soon be ready for implementation within genebanks.

Technical Abstract: Both the United States and Brazil maintain vast collections of grape genetic resources. We share a common interest in using cryopreservation methods for the secure, long-term back-up of accessions within these field collections of the USDA-ARS National Plant Germplasm System and UDESC-CAV Santa Catarina State University in Brazil. We have a developed a Vitis droplet-vitrification method that results in high levels of shoot tip regrowth after liquid nitrogen exposure. Uniform shoot tips were obtained from nodal sections of either in vitro or growth chamber stock plants. Pretreatments included 0.3 M sucrose, salicylic acid, ascorbic acid, and glutathione. Half-strength PVS2 was applied for 30 minutes at 25oC, prior to full-strength PVS2 treatment at 0oC. The optimum PVS2 exposure duration was 30 minutes for growth chamber-derived shoot tips and 90 minutes for in vitro derived shoot tips. Shoot tips were then placed onto foil strips and plunged into LN. Regrowth levels ranged from 45 to 72 % for the seven Vitis species tested from in vitro plants, and from 35 to 65 % for three Vitis vinifera cultivars and one Vitis hybrid tested from growth chamber plants. The high levels of regrowth suggest that Vitis cryopreservation may soon be ready for implementation within genebanks.