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Cryopreservation of in vitro-grown axillary shoot-tip meristems of mint (Mentha spicata L.) by encapsulation vitrification
Authors:D Hirai  A Sakai
Institution:(1) Hokkaido Prefectural Plant Genetic Resources Center, 363-2, Minami-takinokawa, Takikawa, 073-0013 Japan e-mail: hiraidai@agri.pref.hokkaido.jp Fax: +81(0) 125-24-3877, JP;(2) Asabucho 1-5-23, Kitaku, Sapporo 001-0045, Japan, JP
Abstract: Alginate-coated meristems from in vitro-grown axillary buds of mint (Mentha spicata L.) were successfully cryopreserved by vitrification. Excised meristems from nodal segments cold hardened at 4  °C for 3 weeks were encapsulated and osmoprotected by a mixture of 2 M glycerol plus 0.4 M sucrose. These meristems were dehydrated with a highly concentrated vitrification solution (PVS2 solution) for 3 h at 0  °C prior to a plunge into liquid nitrogen. Successfully encapsulated vitrified meristems developed shoots within a week after plating without intermediary callus formation. The average rate of shoot formation amounted to nearly 90%. This procedure was successfully applied to other Mentha species. It was also confirmed that encapsulated vitrified meristems produced a much higher rate of shoot formation than the encapsulated dried meristems. Thus, this revised encapsulation vitrification method appears promising for the cryopreservation of mint and other germplasm. Received: 24 November 1998 / Revision received: 8 February 1999 / Accepted: 26 February 1999
Keywords:  Cryopreservation  Encapsulation-vitrification  Meristems  Mint  Vitrification
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