摘要
【目的】建立适合胡杨休眠茎尖的冷冻保存方法,对珍稀濒危植物胡杨种质资源进行保存。【方法】取深冬胡杨休眠茎尖,无菌处理后,包埋在3%褐藻酸钙胶球中,对包埋后的胶球进行蔗糖预培养,预培养后的胶球再用玻璃化保护液进行处理。研究预培养过程中蔗糖浓度、预培养时间及不同玻璃化保护液和处理时间对胡杨休眠茎尖存活率的影响。【结果】含有休眠茎尖的褐藻酸钙胶球在0.8 mol/L蔗糖溶液中预培养24 h,经玻璃化保护液[40%甘油+45%(0.4 mol/L)蔗糖+10%聚乙二醇(4 000)+5%二甲基亚砜],于室温下处理30 min后再进行冷冻保存,可以获得较好的存活率。【结论】在优化的条件下,胡杨休眠茎尖存活率可高达77%,对保存后再生的幼芽进行遗传稳定性检测,未发现变异。采用液氮冷冻保存胡杨休眠茎尖是可行的。
【Objective】To establish appropriate cryopreservation method to preserve shoot-tips of Populus euphratica dormant buds by using encapsulation-vitrification,thus preserving the germplasm resources of the endangered rare plant.【Method】Dormant shoot-tips were encapsulated in3%calcium alginate beads.The main factors influencing the viability such as sucrose concentration,preculture time,different vitrification solution and the time in vitrification solution were studied.【Result】The results showed that the high viability could be obtained when alginate beads were precultured for24h in the0.8mol/L sucrose and the beads were dehydrated with vitrification solution[40%Gly+45%(0.4mol/L)sucrose+10%PEG(4,000)+5%DMSO]for30min at25℃.【Conclusion】Under optimized conditions,the viability of dormant shoot-tips can reach77%at the most.After cryopreservation,the dormant shoot-tips can grow directly and the genetic stability can be maintained.It is feasible to freeze the dormant shoot tips of Populus euphratica by liquid nitrogen.
作者
张玲
田洪涛
刘锦涛
刘艳萍
ZHANG Ling;TIAN Hong-tao;LIU Jin-tao;LIU Yan-ping(College of Life Science, Tarim University / Xinjiang Production and Construction Corps Key Laboratory)
出处
《新疆农业科学》
CAS
CSCD
北大核心
2017年第12期2232-2238,共7页
Xinjiang Agricultural Sciences
基金
国家自然科学基金项目"超干处理对塔里木盆地四种珍稀濒危植物种子活力影响机理的研究"(31160069)
塔里木大学校长基金"包埋玻璃化法超低温保存塔里木盆地四种珍稀濒危植物的研究"(TDZKSS06001)~~
关键词
胡杨
休眠茎尖
冷冻保存
包埋玻璃化法
Populus euphratica
dormant shoot-tip
cryopreservation
encapsulation-vitrification
