Cryopreservation of potato cultivated varieties and wild species: critical factors in droplet vitrification |
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Authors: | Kim H H Yoon J W P Y E Cho E G Sohn J K Kim T K Engelmann F |
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Affiliation: | National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea. hkim@rda.go.kr |
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Abstract: | The applicability of cryopreservation protocols to a broad range of genotypes is a key issue for genebanks. We tried to identify the critical factors causing differences in survival of cryopreserved shoot tips using potato varieties coming from cultivated and wild species. The droplet-vitrification method, a combination of droplet-freezing and solution-based vitrification, was selected from several protocols. High survival after freezing was observed after dehydration with PVS2 for 20 min, cooling shoot tips placed in a droplet of PVS2 solution on aluminum foil strips by immersing the foil strips in liquid nitrogen, warming them by plunging the foil strips into a 0.8 M sucrose solution (at 40 degrees C) for 30 s and unloading in 0.8 M sucrose for 30 min. This optimized protocol was successfully applied to 12 accessions with survival ranging between 64.0 and 94.4%. |
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