Cryopreservation of in vitro-grown meristems of potato (Solanum tuberosum L.) by encapsulation-vitrification |
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| Authors: | Dai Hirai Akira Sakai |
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| Institution: | (1) Hokkaido Prefectural Plant Genetic Resources Center, 363-2, Minami-takinokawa, 073-0013 Takikawa, Hokkaido, Japan;(2) 1-5-23, Asabu-cho, Kita-ku, 001-0045 Sapporo, Hokkaido, Japan |
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| Abstract: | Summary Alginate coated meristems from in vitro-grown axillary buds of potato (Solanum tuberosum L.) were successfully cryopreserved by vitrification. Excised meristems were precultured on sucrose-enriched MS medium and
then encapsulated. To induce dehydration tolerance (osmotolerance), encapsulated meristems were treated with a mixture of
2 M glycerol plus 0.6 M sucrose for 90 min. These encapsulated meristems were dehydrated with a highly concentrated vitrification
solution (PVS2 solution) for 3 hr at 0°C prior to a plunge into liquid nitrogen. Successfully vitrified meristems developed
shoots within 3 weeks after plating without intermediary callus formation. The average rate of shoot formation amounted to
nearly 70%. No difference was observed in RAPD analysis using 17 primers between cryopreserved and non-treated plantlets.
The cryogenic protocol was successfully applied to 14 cultivars. It was also confirmed that the encapsulated vitrified meristems
produced much greater shoot formation than the encapsulated dried meristems. Thus, the encapsulation vitrification protocol
appears promising for cryopreservation of potato germplasm. |
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| Keywords: | Osmoprotection |
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