[an error occurred while processing this directive]

Plant Diversity ›› 2015, Vol. 37 ›› Issue (06): 801-812.DOI: 10.7677/ynzwyj201515067

• 研究论文 • 上一篇    下一篇

红芽芋茎尖的包埋玻璃化法超低温保存

 王艾平, 尹明华, 洪森荣   

  1. 上饶师范学院生命科学学院,江西 上饶334001
  • 收稿日期:2015-04-22 出版日期:2015-11-25 发布日期:2015-06-23
  • 基金资助:

    Science and Technology Ground Plan Project of College and University in Jiangxi Province in 2013 (KJLD13088); Agricultural Science and Technology Support Program of Science & Technology Department of Jiangxi Province (20122BBF60126) and National Natural Science Fund in China (31360072)

Cryopreservation of in vitroGrown Shoot Tips of Red Bud Taro by Encapsulation-Vitrification

 WANG  Ai-Ping, YIN  Ming-Hua, HONG  Sen-Rong   

  1. College of Life Sciences, Shangrao Normal University, Shangrao, Jiangxi 334001, China
  • Received:2015-04-22 Online:2015-11-25 Published:2015-06-23
  • Supported by:

    Science and Technology Ground Plan Project of College and University in Jiangxi Province in 2013 (KJLD13088); Agricultural Science and Technology Support Program of Science & Technology Department of Jiangxi Province (20122BBF60126) and National Natural Science Fund in China (31360072)

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: