吉林农业大学学报2025，Vol.47Issue(1)：49-58,10.DOI:10.13327/j.jjlau.2021.1098
蓝果忍冬离体茎尖小滴玻璃化超低温保存体系优化
Optimization of Cryopreservation System of Shoot Tips in Vitro of Blue Honeysuckle(Lonicera L.subsect.Caeruleae)by Droplet Vitrifi-cation
摘要
Abstract
The shoot tips of test tube plantlets of blue honeysuckle"Огненный опал"were cryopre-served by droplet vitrification method.The method was verified by 5 genotypes.The results of param-eter optimization study showed that stem segments of 1.5 cm in length with terminal buds were cut from 4-week-old test tube plantlets and cultured in 4℃and the dark for 15 days.2-3 mm long shoot tips of terminal buds were cultured on preculture medium(MS+0.5 mol/L sucrose+6.5 g/L agar)for 2 days[(25±2)℃,dark].The shoot tips were put into loading medium(MS+2.0 mol/L glyc-erol+0.4 mg/L sucrose)for 40 min(room temperature).After that,The shoot tips were treated with PVS2 for 40 min(0℃).The shoot tips were then transferred to the droplets prepared with PVS2 on the aluminum foil(0℃)and quickly put into liquid nitrogen(LN)for 1 h.The material taken out from LN was quickly put into unloading solution(MS+1.2 mol/L sucrose)for 30 min(room temperature).Finally,the shoot tips were cultured on the regeneration medium(MS+1.0 mg/L 6-BA+0.3 mg/L IBA+30 g/L sucrose+6.5 g/L agar)for 5 days[(25±2)℃,dark]and then cultured under normal temperature and light.The average regeneration rate of the 5 genotypes after cryopreservation was(44.7±5.0)%.The highest regeneration rate was(55.0±5.4)%.The droplet vitrification cryopreser-vation system for blueberry honeysuckle germplasm resources provides a new method for the long-term and effective preservation of blueberry honeysuckle germplasm resources.关键词
蓝果忍冬/离体培养/小滴玻璃化/超低温保存Key words
blue honeysuckle/culture in vitro/droplet vitrification/cryopreservation分类
农业科技引用本文复制引用
梁杉,张志东,刘海广,陈丽,李亚东,李金英..蓝果忍冬离体茎尖小滴玻璃化超低温保存体系优化[J].吉林农业大学学报,2025,47(1):49-58,10.基金项目
吉林省科技发展计划项目(20180201076NY,20200402080NC),吉林省"三区"人才支持计划项目(202022309) （20180201076NY,20200402080NC）
