九里香种子超低温保存研究  

Study on Cryopreservation of Murraya exotica L.Seeds

作　　者：顾雅坤 吴怡[1] 符丽 郑希龙[1] 曾琳[1] 魏建和[1,2] GU Ya-kun;WU Yi;FU Li;ZHENG Xi-long;ZENG Lin;WEI Jian-he(Chinese Academy of Medical Sciences&Peking Union Medical College/Hainan Branch Institute of Medicinal Plant Development/Hainan Provincial Key Laboratory of Resources Conservation and Development of Southern Medicine,Haikou 570311;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193)

机构地区：[1]中国医学科学院北京协和医学院/药用植物研究所海南分所/海南省南药资源保护与开发重点实验室,海口570311 [2]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《植物遗传资源学报》2020年第3期760-766,共7页Journal of Plant Genetic Resources

基　　金：国家自然科学基金青年基金项目(81603399);中国医学科学院医学与健康科技创新工程项目(2016-l2M-2-003)。

摘　　要：研究旨在探索适于九里香(Murraya exotica L.)种子的液氮超低温保存条件,为采用液氮超低温保存方法长期保存九里香种子提供依据。以九里香成熟种子为材料,采用快速干燥法获得不同含水量的种子,分别用玻璃化法、分步冷冻法和直接冷冻法对其进行液氮超低温保存,结合石蜡切片法观察种子显微性状,并测定种子生理生化活性指标。结果发现,种子含水量由30%降至7%,其对照组生活力由98%缓慢降至82%;经不同冷冻方法处理后其生活力随含水量的降低呈先降后升的趋势,在含水量7%时各冷冻组生活力均升至最高,且此时冻存前后生活力最接近;分步冷冻处理后的种子生活力普遍高于玻璃化冷冻和直接冷冻处理;含水量18%~30%时种子分步冷冻处理后切面颜色较其他组偏深,含水量30%的种子分步冷冻处理后部分细胞出现空腔或较严重的质壁分离,含水量7%的种子经不同冷冻处理后其活力和结构与对照组相比基本无变化;分步冷冻处理后种子丙二醛含量、蛋白质含量、过氧化氢酶活性、过氧化物酶活性下降,脱氢酶活性升高。九里香种子可用液氮超低温保存,采用含水量为7%的种子分步冷冻处理为佳。This study attempted to explore the suitable liquid nitrogen cryopreservation condition of Murraya exotica L.seed and provided a pipeline for the long-term preservation of the seeds.Using mature seeds of Murraya exotica L.with different water content using rapid drying method,cryopreservation experiments were carried out with three approaches including vitrification method,step freezing and direct freezing method,followed by an observation of the micro-characterswith the method of paraffin section and the quantification of the physiological and biochemical activity indexes.The results showed that the water content of the seed decreased from 30%to 7%,and the viability of the control group slowly decreased from 98%to 82%.After being treated by different freezing methods,the viability first decreased and then rised with the decrease of water content.At 7%water content,the viability of each freezing group reached to the highest,and the viability is the closest before and after cryopreservation at this time.The seed vigor after step freezing was generally higher than that of vitrification and direct freezing.Among them,the seed color at the cutting surface were darker than other groups after step freezing,and the seeds with 30%water content had cavity or severe plasmolysis in some cells after step freezing.The vigor and structure of the seeds in the 7%water content group were basically unchanged after the different freezing treatments compared with the control group.After the step freezing treatment,the malondialdehyde content,protein content,catalase activity and peroxidase activity of the seeds decreased and the dehydrogenase activity was found to be elevated.Thus,we concluded that the Murraya exotica L.seeds are able to be stored following the ultralow temperature preservationpipeline in liquid nitrogen,by use of the water content of 7%seed with themethod of step freezing.

关 键 词：九里香种子 液氮超低温保存 含水量 冷冻方法 解冻方法 

分 类 号：S685.99[农业科学—观赏园艺]