胡杨休眠茎尖的包埋玻璃化法冷冻保存研究

doi:10.6048/j.issn.1001-4330.2017.12.010

新疆农业科学 ›› 2017, Vol. 54 ›› Issue (12): 2232-2238.DOI: 10.6048/j.issn.1001-4330.2017.12.010

胡杨休眠茎尖的包埋玻璃化法冷冻保存研究

张玲, 田洪涛, 刘锦涛, 刘艳萍

塔里木大学生命科学学院/新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,新疆阿拉尔 843300

收稿日期:2017-09-08 出版日期:2017-12-25 发布日期:2018-01-29
通信作者: 刘艳萍(1977-),女,辽宁沈阳人,副教授,硕士,研究方向为植物种质保存,(E-mail)13785186@qq.com
作者简介:张玲(1977-),女,新疆石河子人,副教授,硕士,研究方向为资源植物学,(E-mail)185499412@qq.com
基金资助:
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)

Cryopreservation of Shoot-tips of Populus euphratica Dormant Buds by Encapsulation-vitrification

ZHANG Ling, TIAN Hong-tao, LIU Jin-tao, LIU Yan-ping

College of Life Science, Tarim University / Xinjiang Production and Construction Corps Key Laboratory Biological Resources Conservation and Utilization in Tarim Basin, Tarim University,Alar Xinjiang 843300, China

Received:2017-09-08 Online:2017-12-25 Published:2018-01-29
Correspondence author: Liu Yan-ping,Female(1977-),Native place:Shenyang,Liaoning Province,associate professor,research field: Plant germplasm conservation,(E-mail)13785186@qq.com
Supported by:
National Natural Science Foundation of China"Study on the mechanism of ultra-dry preservation on seed vigor of four rare and endangered plants in Tarim Basin(31160069)";Tarim University president fund"Study on cryopreservation of four rare and endangered plants in Tarim Basin by vitrification(TDZKSS06001)"

摘要/Abstract

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。

关键词: 胡杨; 休眠茎尖; 冷冻保存; 包埋玻璃化法

Abstract: 【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 in 3% 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 for 24 h in the 0.8 mol/L sucrose and the beads were dehydrated with vitrification solution [40% Gly + 45% (0.4 mol/L) sucrose + 10% PEG (4,000) + 5% DMSO] for 30 min at 25℃. 【Conclusion】Under optimized conditions, the viability of dormant shoot-tips can reach 77% 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.

Key words: Populus euphratica; dormant shoot-tip; cryopreservation; encapsulation-vitrification

中图分类号:

S722

张玲, 田洪涛, 刘锦涛, 刘艳萍. 胡杨休眠茎尖的包埋玻璃化法冷冻保存研究[J]. 新疆农业科学, 2017, 54(12): 2232-2238.

ZHANG Ling, TIAN Hong-tao, LIU Jin-tao, LIU Yan-ping. Cryopreservation of Shoot-tips of Populus euphratica Dormant Buds by Encapsulation-vitrification[J]. Xinjiang Agricultural Sciences, 2017, 54(12): 2232-2238.

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胡杨休眠茎尖的包埋玻璃化法冷冻保存研究

Cryopreservation of Shoot-tips of Populus euphratica Dormant Buds by Encapsulation-vitrification

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