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Cryopreservation of shoot tips of endangered Hayachine-usuyukiso (Leontopodium hayachinense (Takeda) Hara et Kitam.) using a vitrification protocol

Published online by Cambridge University Press:  15 May 2008

Daisuke Tanaka
Affiliation:
Genebank, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
Takao Niino*
Affiliation:
Genebank, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
Yoshiko Tsuchiya
Affiliation:
Graduate School of Agriculture, Iwate University, Morioka 020-8550, Japan
Kazuto Shirata
Affiliation:
Genebank, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
Matsuo Uemura
Affiliation:
Graduate School of Agriculture, Iwate University, Morioka 020-8550, Japan
*
*Corresponding author. E-mail: [email protected]

Abstract

Hayachine-usuyukiso (Leontopodium hayachinense) is an alpine plant native to Mt Hayachine. This unique chrysanthemum is listed as an endangered plant by the Department of Conservation, Iwate Prefecture, and as a threatened plant by the Ministry of the Environment, Japan. We successfully cryopreserved the shoot tips from in vitro-grown L. hayachinense shoots using a vitrification protocol. Cold-hardened shoot tips were excised and pre-cultured on a solidified Murashige–Skoog medium containing 0.3 M sucrose for 1 d at 5°C. The shoot tips were then treated with loading solution for 20 min at 25°C, dehydrated in plant vitrification solution 2 for 120 min at 25°C and immersed in liquid nitrogen. The survival rate of the vitrified shoot tips was 63.3% after 30 d of regrowth. This protocol appears to be a promising technique for the cryopreservation of in vitro-grown shoots of this endangered plant.

Type
Short Communication
Copyright
Copyright © NIAB 2008

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