Articles
IMPLEMENTATION OF CRYOPRESERVATION FOR GARLIC GENETIC RESOURCES BY THE DROPLET VITRIFICATION PROCEDURE
Article number
760_27
Pages
209 – 215
Language
English
Abstract
The droplet-vitrification protocol, a combination of droplet-freezing and solution-based vitrification was applied for cryopreserving garlic bulbil primordia.
The highest regrowth of cryopreserved primordia (82.7-85.0 %) was achieved after preculture for 2-4 days at 10°C on solid medium with 0.1-0.3 M sucrose, loading for 50 min in liquid medium with 2 M glycerol + 0.5 M sucrose, dehydration with PVS3 solution for 90-150 min, cooling primordia in 5 µl droplets of PVS3 placed on aluminum foil strips, warming them by plunging the foil strips into 40°C, 0.8M sucrose solution.
The optimized droplet-vitrification protocol was successfully applied to bulbil primordia of five garlic varieties originating from various countries and to immature bulbils of two vegetatively propagated Allium species, with regrowth percentages ranging between 77.4-95.4 %. This protocol was further applied to the Korean garlic collection for safety back up of germplasm conservation.
A total of 351 accessions were tested (with regrowth of 75.6-84.7%) and, among them, 260 accessions of three clonal Allium species were stored in liquid nitrogen.
The highest regrowth of cryopreserved primordia (82.7-85.0 %) was achieved after preculture for 2-4 days at 10°C on solid medium with 0.1-0.3 M sucrose, loading for 50 min in liquid medium with 2 M glycerol + 0.5 M sucrose, dehydration with PVS3 solution for 90-150 min, cooling primordia in 5 µl droplets of PVS3 placed on aluminum foil strips, warming them by plunging the foil strips into 40°C, 0.8M sucrose solution.
The optimized droplet-vitrification protocol was successfully applied to bulbil primordia of five garlic varieties originating from various countries and to immature bulbils of two vegetatively propagated Allium species, with regrowth percentages ranging between 77.4-95.4 %. This protocol was further applied to the Korean garlic collection for safety back up of germplasm conservation.
A total of 351 accessions were tested (with regrowth of 75.6-84.7%) and, among them, 260 accessions of three clonal Allium species were stored in liquid nitrogen.
Authors
K. Haeng-Hoon, L. Yun-Keol, K. Taesan, L. Joung-Kwan, J. Jae-Jun, N. Sang-Sik, F. Engelmann
Keywords
Allium sativum L., bulbil primordium, cooling, warming, PVS3 solution
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