摘要
利用流式细胞技术对金线莲基因组大小进行测定,选取金线莲新鲜嫩叶为待测材料,选用3种不同的缓冲液进行解离,制备细胞核悬浮液.以鸡红细胞核为内标,使用碘化丙啶(PI)染色,通过流式细胞仪对其荧光强度进行测定;通过比较换算,建立了以流式细胞技术测定金线莲基因组大小的参考方法.研究发现使用Galbraith′s解离液对金线莲样品的解离效果较好,测得金线莲基因组DNA的2C含量为6.83±0.067 pg.结果表明,基于流式细胞技术,优化解离和测定方法可准确测定金线莲的基因组大小.
To determine the genome size of Anoectochilus roxburghii by flow cytometry method,fresh leaves of A.roxburghii were collected and dissociated by three different buffer to extract their nuclei.Chicken erythocyte nuclei was used as the internal standard and propidium iodide was used as the staining material.Based on fluorescence intensities measured by flow cytometer,the genome size of A.roxburghii was estimated referring to the internal standard.The results showed that Galbraith′s buffer was the most suitable solution for A.roxburghii dissociation,and the 2 C value of A.roxburghii.DNA was(6.83±0.067)pg.The study verified that the method based on flow cytometry analysis can accurately estimate the genome size of A.roxburghii.
作者
林瀚
王晶晶
黄华星
何碧珠
兰思仁
马晓开
LIN Han;WANG Jingjing;HUANG Huaxing;HE Bizhu;LAN Siren;MA Xiaokai(Center for Genomics and Biotechnology,Fujian Agriculture and Forestry University;Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization(Fuzhou);College of Landscape Architecture,Fujian Agriculture and Forestry University;College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2020年第6期766-771,共6页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
国家自然科学基金青年科学基金(31700199)。
关键词
金线莲
流式细胞技术
基因组大小
Anoectochilus roxburghii
flow cytometry
genome size
