红芽芋茎尖的包埋玻璃化法超低温保存

doi:10.7677/ynzwyj201515067

摘要/Abstract

摘要：

对红芽芋（Colocasia esculenta var. cormosus ‘Hongyayu’）茎尖的包埋玻璃化法超低温保存技术进行了研究。茎尖从培养8周的试管苗上切下并包埋成海藻酸钙凝胶珠，并在MS+35mg·L-1 6BA+05mg·L-1 IBA+ 01mg·L-1 GA3 +03mol·L-1蔗糖的液体培养基中预培养24h，随后用2mol·L-1甘油+04mol·L-1蔗糖的混合物在25℃下装载30min，并用PVS2在25℃脱水20min后将包埋的茎尖直接投入液氮保存。保存1d后取出材料在40℃水浴快速复温3min后，吸去冷冻管中PVS2，并用MS+35mg·L-1 6BA+05mg·L-1 IBA+01mg·L-1 GA3+12mol·L-1蔗糖的液体培养基在25℃洗涤3次，每次10min。最后将茎尖接种于MS+35mg·L-1 6BA+05mg·L-1 IBA+01mg·L-1 GA3的固体培养基上，暗培养3d后转入正常的光周期中培养。红芽芋茎尖冻后成活率约为80%，其再生植株没有发生形态学的变化。这种包埋玻璃化法程序有望成为红芽芽茎尖超低温保存的常规方法。

关键词: 超低温保存, 包埋玻璃化法, 红芽芋, 茎尖

Abstract:

A simple procedure for cryopreservation of in vitrogrown shoot tips of red bud taro (Colocasia esculenta L. Schott var. cormosus‘Hongyayu’) by encapsulationvitrification is investigated. Shoot tips were excised from 8weekold stock shoots and encapsulated into alginategel beads. Encapsulated shoot tips were precultured in liquid MS medium supplemented with 35mg·L-1 6BA, 05mg·L-1 IBA, 01mg·L-1 GA3 and 03mol·L-1 sucrose for 24h, then they were loaded with a mixture of 2mol·L-1 glycerol plus 04mol·L-1 sucrose for 30min at 25℃. After dehydration with PVS2 at 25℃ for 20min, the encapsulated and dehydrated shoot tips were plunged directly into liquid nitrogen. After rapidly rewarming in a 40℃ water bath for 3min, PVS2 was drained from the cryotubes and replaced third with liquid MS medium supplemented with 35mg·L-1 6BA, 05mg·L-1 IBA, 01mg·L-1 GA3 and 12mol·L-1 sucrose and each kept for 10min at 25℃and then postcultured on solidified MS medium supplemented with 35mg·L-1 6BA, 05mg·L-1 IBA and 01mg·L-1 GA3 in the dark for 3 days and then transferred to the light conditions. The average survival rate amounted to about 80%. Plantlets regenerated from cryopreserved shoot tips were morphologically uniform. This encapsulation vitrification procedure promises to become a routine method for the cryopreservation of shoot tips of Chinese genuine red bud taro.

Key words: Cryopreservation, Encapsulation-vitrification, Red bud taro (Colocasia esculenta var. cormosus‘Hongyayu’), Shoot-tips

中图分类号:

Q 943，S 632.3

王艾平, 尹明华, 洪森荣. 红芽芋茎尖的包埋玻璃化法超低温保存[J]. Plant Diversity, 2015, 37(06): 801-812.

WANG Ai-Ping, YIN Ming-Hua, HONG Sen-Rong. Cryopreservation of in vitroGrown Shoot Tips of Red Bud Taro by Encapsulation-Vitrification[J]. Plant Diversity, 2015, 37(06): 801-812.

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