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采用包埋/脱水法对体外培养的啤酒花(Humulus lupulus L.)茎尖进行超低温保存。

Cryopreservation of in vitro-grown shoot-tips of hop (Humulus lupulus L.) using encapsulation/dehydration.

作者信息

Martínez D, Tamés R S, Angeles Revilla M

机构信息

Unidad de Fisiología Vegetal, Dpto. Biología de Organismos y Sistemas, Universidad de Oviedo, 33005 Oviedo, Spain Fax: +34-85-5104867, , , , , , ES.

出版信息

Plant Cell Rep. 1999 Nov;19(1):59-63. doi: 10.1007/s002990050710.

Abstract

A cryopreservation procedure using encapsulation/dehydration was established for shoot-tips obtained from in vitro-grown shoots of hop. After dissection, shoot-tips were encapsulated in medium with alginate and 0.5 M sucrose. Optimal conditions consisted of preculture for 2 days in solid medium with 0.75 M sucrose, or in increasing sucrose concentrations, desiccation for 4 h with silicagel in a flow cabinet (16% water content) followed by rapid freezing and slow thawing. Shoot recovery after freezing 60 min in liquid nitrogen was around 80%. No phenotypical changes were observed in the recovered plants from cryopreserved shoot-tips growing in the field.

摘要

建立了一种使用包封/脱水的冷冻保存方法,用于从啤酒花离体培养芽中获取的茎尖。解剖后,将茎尖包封在含有藻酸盐和0.5 M蔗糖的培养基中。最佳条件包括在含有0.75 M蔗糖的固体培养基中预培养2天,或在逐渐增加的蔗糖浓度下预培养,在通风橱中用硅胶干燥4小时(含水量16%),然后快速冷冻和缓慢解冻。在液氮中冷冻60分钟后的茎尖恢复率约为80%。从冷冻保存的茎尖在田间生长的再生植株中未观察到表型变化。

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