种子2025，Vol.44Issue(5)：16-25,35,11.DOI:10.16590/j.cnki.1001-4705.2025.05.016
米槁胚性愈伤组织玻璃化法超低温保存体系的建立
Contruction of Vitrification-based Cryopreservation System for the Embryogenic Callus of Cinnamomum migao
摘要
Abstract
In order to establish a cryopreservation system for embryogenic callus of Cinnamomum mi-gao and solve the problem of long-term preservation difficulty of embryogenic callus germplasm re-sources,the Cinnamomum migao embryogenic callus was used as the material and cryopreservation was carried out by vitrification method.The effects of the type of cryoprotectant and dehydration time on the survival rate,regeneration rate and recovery ability of embryogenic callus were studied,and regeneration rate and recovery ability of embryonic callus of ricegeria were tested.The results showed that the suitable cryoprotectant for embryonic callus was PVS2.The cell survival rate was the highest after 70 minutes of PVS2 treatment,which was 80.21％.The suitable cryopreservation method for embryonic callus was Loading(WPM+2 mol/L glycerol+0.4 mol/L sucrose)at 25℃ for 20 minutes,PVS2(WPM+30％glycerol+0.4 mol/L sucrose+15％ethylene glycol+15％dimethyl sulfoxide)at 0℃ for 10 minutes,thawed in a 40℃ water bath for 2 minutes,unloading(WPM+1.2 mol/L su-crose)at 25℃ for 20 minutes.Except for the dehydration treatment of PVS2 for 0 minute,a 100％re-generation rate could be achieved under the remaining treatments.The differentiation ratios of cotyle-donous embryos and mature embryos after cryopreservation were both higher than those of embryonic callus that was not cryopreserved.There was no difference in the morphology of the somatic embryos differentiated from embryonic callus that was cryopreserved and that of embryonic callus that was not cryopreserved,indicating that embryonic callus could be used as ultra-low temperature cryopreserva-tion material for the germplasm resources of Cinnamomum migao for long-term ultra-low tempera-ture cryopreservation.关键词
米槁/胚性愈伤组织/超低温保存/玻璃化法Key words
Cinnamomum migao/embryogenic callus/cryopreservation/vitrification method分类
农业科技引用本文复制引用
陈宇凡,吴媛,林亮,杨湘云,帅甜,丁宁,吴之坤..米槁胚性愈伤组织玻璃化法超低温保存体系的建立[J].种子,2025,44(5):16-25,35,11.基金项目
贵州省科技计划项目(黔科中引地[2022]4016) （黔科中引地[2022]4016）
