文献类型：期刊文献

中文题名：黄芪休眠芽茎尖玻璃化超低温保存研究

英文题名：Study on vitrification cryopreservation of dormant buds of Astragalus membranaceus(Fisch)Bge.var.mongholicus(Bge)Hsiao

作者：董婉琦[1];张延红[1];何春雨[1];唐顺莉[1]

第一作者：董婉琦

机构：[1]甘肃中医药大学药学院,甘肃兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2022

卷号：33

期号：6

起止页码：1463

中文期刊名：时珍国医国药

外文期刊名：Lishizhen Medicine and Materia Medica Research

收录：北大核心:【北大核心2020】；CSCD:【CSCD_E2021_2022】；

基金：国家自然科学基金(81960683);甘肃省高等学校产业支撑计划项目(2020C-09);甘肃省教育厅青年博士基金项目(2021QB-073);甘肃省民生科技专项(科技特派员专题)(20CX9NA070);中央引导地方科技发展专项资金项目(30440323);现代农业产业技术体系建设专项(CARS-21);甘肃省创新基地人才计划(18JR2TA017)。

语种：中文

中文关键词：黄芪;休眠芽;玻璃化超低温保存;植株再生

外文关键词：Astragalus membranaceus var.mongholicus;Dormant buds;Vitrification cryopreservation;Plant regeneration

摘要：目的建立黄芪休眠芽茎尖玻璃化超低温保存技术体系。方法采用单因子实验设计,研究PVS2处理时间、休眠芽大小、冻存茎尖大小以及缺少装载和PVS2处理对黄芪休眠芽玻璃化超低保存成活率的影响。结果将一年生黄芪种苗覆土后置于4℃低温锻炼20d左右,切取完整的休眠芽,剥至1~2 mm长的茎尖,先用含2 mol/L甘油+0.4 mol/L蔗糖的MS溶液装载20 min,再用PVS2处理50 min后投氮,恢复培养时从液氮中取出,38℃水浴化冻2 min,用无菌滤纸吸干表面溶液后接种到恢复培养基中(MS+2.0 mg/L BA+0.5 mg/L IAA+3%蔗糖+0.7%琼脂,pH5.8),先暗培养一周,再移到正常光照下进行培养,休眠芽冻存后的成活率高达100%。结论建立了高效的黄芪休眠芽玻璃化超低温保存技术体系,为药用植物的种质资源保存开辟了一条新的途径。
Objective To establish the vitrification cryopreservation system of the dormant buds of Astragalus membranaceus var.mongholicus.Methods This paper dealt with the effect of PVS2 treatment time,bud size,stem tip size and with or without PVS2 and loading treatments on surviving rate using single factor experiment design.Results One year old fresh Astragalus membranaceus var.mongholicus was covered with soil and kept at 4℃for about 20 days.Complete dormant buds with base were cut and peeled to 1~2 mm stem tips which were firstly loaded with MS medium containing 2 mol/L glycerin and 0.4 mol/L sucrose for 20 min,and then treated with PVS2 for 50 min and finally put into nitrogen.When the stem tips need recovery culture,they were thawed in 38℃water bath for 2 min and blotted the surface solution with sterile filter paper and inoculated it into medium(MS+2.0 mg/L BA+0.5 mg/L IAA+3%sucrose+0.7%agar,pH5.8).The survival rate of dormant buds after cryopreservation was 100%.Conclusion An efficient vitrification cryopreservation technology system of Astragalus membranaceus dormant buds is established,which could provide a new approach to the conservation of medicinal plant germplasm resources.