甘肃贝母休眠芽玻璃化超低温保存过程中的淀粉粒变化研究  

Study on the changes of starch granules in dormant buds of Fritillaria przewalskii when cryo⁃preserved by vitrification

作　　者：何玉明 张延红 何春雨[1,2,3] 郭清毅 HE Yu-ming;ZHANG Yan-hong;HE Chun-yu;GUO Qing-yi(School of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,China;Collaborative Innovation Center of Natural Medicine Germplasm Resources Discovering and New Cultivars Breeding,Lanzhou 730000,China;Xinglin Baicao Garden of Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学药学院,甘肃兰州730000 [2]天然药用种质资源挖掘与新品种选育协同创新中心,甘肃兰州730000 [3]甘肃中医药大学杏林百草园,甘肃兰州730000

出　　处：《时珍国医国药》2025年第4期744-750,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81960683);甘肃省自然科学基金项目(21JR1RA263);兰州市科技计划项目(2019-1-83,2020-ZD-59);甘肃中医药大学硕士研究生创新创业基金项目。

摘　　要：目的从组织学层面揭示甘肃贝母休眠芽在应对玻璃化超低温保存逆境时淀粉粒做出的响应,为阐明甘肃贝母休眠芽玻璃化超低温保存机理提供理论依据。方法以甘肃贝母休眠芽为试验材料,采用石蜡制片技术、通过PAS染色(多糖染色)研究了休眠芽在玻璃化超低温保存过程中装载液、PVS2玻璃化保护液、液氮冻存与化冻及恢复培养这几个环节对细胞中淀粉粒的影响,以不经处理的新鲜休眠芽为对照。结果在超低温保存过程中,顶端分生组织能够被完整地保留下来,而最外层叶原基和芽轴会受到严重的伤害。结论在应对玻璃化超低温保存这种逆境胁迫的过程时,细胞中淀粉粒数量的增加、形态的变大变圆是抵抗逆境,保护细胞免受冻害的重要措施。Objective To reveal the response of starch granules in dormant buds of Fritillaria przewalskii Maxim.ex Batal.(F.przewal⁃skii)to the stress of cryopreservation by vitrification from the histological level,and to provide a theoretical basis for elucidating the mechanism of vitrification cryopreservation of dormant buds of F.przewalskii.Methods Dormant buds of F.przewalskii were used as ex⁃perimental materials.The effects of loading solution,PVS2 vitrification solution,liquid nitrogen freezing and thawing and recovery cul⁃ture on starch granules in cells during vitrification cryopreservation of dormant buds were studied by paraffin section technology and peri⁃odic acid-schiff(PAS)staining(polysaccharide staining).The fresh dormant buds without treatment were used in the control group.Results The apical meristem remained fully intact after cryopreservation,while severe structural damage was observed in the outermost leaf primordium and bud axis.Conclusion In the process of responding to the adversity stress of vitrification cryopreservation,the in⁃crease in the quantity of starch granules and the enlargement and roundness of the morphology are critical adaptive responses to vitrifica⁃tion cryopreservation stress,serving as key mechanisms to mitigate freezing injury and enhance cellular protection.

关 键 词：甘肃贝母 玻璃化超低温保存 淀粉粒分布 冻存机理 

分 类 号：S567[农业科学—中草药栽培]