Cryopreservation of 12 Vitis Species Using Apical Shoot Tips Derived from Plants Grown In Vitro

in HortScience

The availability of and easy access to diverse Vitis species are prerequisites for advances in breeding programs. Plant genebanks usually maintain collections of Vitis taxa as field collections that are vulnerable to biotic and abiotic stresses. Cryopreservation has been considered an ideal method of preserving these collections as safety back-ups in a cost-effective manner. We report a droplet vitrification method used to cryopreserve 12 Vitis species (Vitis vinifera cvs. Chardonnay and ‘Riesling, V. actinifolia, V. aestivalis, V. jacquemontii, V. flexuosa, V. palmata, V. riparia, V. rupestris, V. sylvestris, V. ficifolia, V. treleasi, and V. ×novae angeliae) using shoot tips excised from plants grown in vitro. Our results demonstrated wide applicability of this technique, with regrowth levels at least 43% for 13 genotypes representing 12 Vitis species. We demonstrated that the droplet vitrification procedure can be successfully replicated by technical staff, thus suggesting that this method is ready for implementation.

Contributor Notes

This work was supported in part by the USDA-ARS National Laboratory for Genetic Resources Preservation) and The Science and Technology Development Fund (STDF/Egypt).

The authors are grateful to the Coordination for the Improvement of Higher Education Personnel (CAPES) of the Brazilian Ministry of Education (MEC) for the PhD Scholarship in Brazil and the Scholarship for the PhD Sandwich Program Abroad (PDSE) in the United States of America granted to Jean Carlos Bettoni to perform research at the USDA-ARS National Laboratory for Genetic Resources Preservation (NLGRP). Research was funded in part by NAS and USAID, and any opinions, findings, conclusions, or recommendations expressed are those of the authors alone and do not necessarily reflect the views of USAID or NAS. The authors thank Ben Gutierrez for providing an internal review of the manuscript.

Corresponding author. E-mail: gayle.volk@ars.usda.gov.

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    Grapevine plants recovered after 90 min of plant vitrification solution 2 (PVS2) treatment with liquid nitrogen (+LN) or without liquid nitrogen (−LN) exposure. V. aestivalis shoot tips exposed to LN and recovered for 30 d (A) and 2 months (B) and without LN exposure after 2 months of regrowth (C). V. jacquemontii shoot tips exposed to LN and recovered for 30 d (D) and 2 months (E) and without LN exposure after 2 months of regrowth (F). ‘Chardonnay’ shoot tips exposed to LN and recovered for 30 d (G) and 2 months (H) and without LN exposure after 2 months of regrowth (I). V. flexuosa shoot tips exposed to LN and recovered for 30 d (J) and 2 months (K) and without LN exposure after 2 months of regrowth (L). V. palmata shoot tips exposed to LN and recovered for 42 d (M) and 2 months (N) and without LN exposure after 2 months of regrowth (O). Scale bars: A = 1 mm; D, G, J, M = 2 mm.

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