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+3.5 mg·L^(-1)6-BA+0.5 mg·L^(-1)IBA+0.1 mg·L^(-1)GA_3+0.3 mol·L^(-1)蔗糖的液体培养基中预培养24 h,随后用2 mol·L^(-1)甘油+0.4 mol·L^(-1)蔗糖的混合物在25℃下装载30 min,并用PVS2在25℃脱水20 min后将包埋的茎尖直接投入液氮保存。保存1 d后取出材料在40℃水浴快速复温3 min后,吸去冷冻管中PVS2,并用MS+3.5 mg·L^(-1)6-BA+0.5mg·L^(-1)IBA+0.1 mg·L^(-1)GA_3+1.2 mol·L^(-1)蔗糖的液体培养基在25℃洗涤3次,每次10 min。最后将茎尖接种于MS+3.5 mg·L^(-1)6-BA+0.5 mg·L^(-1)IBA+0.1 mg·L^(-1)GA_3的固体培养基上,暗培养3 d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。
A simple procedure for cryopreservation of in vitro-grown shoot tips of red bud taro （ Colocasia esculenta L. Schott var. cormosusHongyayu＇ ） by encapsulation-vitrification is investigated. Shoot tips were excised from 8- week-old stock shoots and encapsulated into alginate-gel heads. Encapsulated shoot tips were precultured in liquid MS medium supplemented with 3.5 mg.L -1 6-BA, 0. 5 mg.L -1 IBA, 0. 1 mg. L-1 GA3 and 0. 3 mol.L -1 sucrose for 24 h, then they were loaded with a mixture of 2 mol. L-1 glycerol plus 0. 4 mol. L -1 sucrose for 30 rain at 25 ℃. After dehydration with PVS2 at 25 ℃ for 20 min, the encapsulated and dehydrated shoot tips were plunged directly into liquid nitrogen. After rapidly rewarming in a 40 ℃ water bath for 3 rain, PVS2 was drained fiom the cryotubes and replaced third with liquid MS medium supplemented with 3.5 mg. L-1 6-BA, 0. 5 mg. L-1 IBA, 0. 1 mg. L -1 GA3 and 1.2 mol. L-1 sucrose and each kept for 10 rain at 25 ℃and then post-cultured on solidified MS medium supple- mented with 3.5 mg. L-1 6-BA, 0. 5 mg. L-1 IBA and 0. 1 mg. 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.
出处
《植物分类与资源学报》
CAS
CSCD
北大核心
2015年第6期801-812,共12页
Plant Diversity
基金
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)
National Natural Science Fund in China(31360072)
关键词
超低温保存
包埋玻璃化法
红芽芋
茎尖
Cryopreservation
Encapsulation-vitrification
Red bud taro （ Colocasia esculenta var. cormosus ‘ Hongy-ayu＇ ）
Shoot-tips
