流式细胞术测定苎麻基因组大小  

Determination of Ramie(Boehmeria nivea) Genome Size by Flow Cytometry

作　　者：何鑫玺 钟启 钟英丽[1] 揭雨成[2] He Xinxi;Zhong Qi;Zhong Yingli;Jie Yucheng(College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha,410000;College of Agronomy,Hunan Agricultural University, Changsha,410000)

机构地区：[1]湖南农业大学生物科学技术学院,长沙410000 [2]湖南农业大学农学院,长沙410000

出　　处：《分子植物育种》2019年第1期99-103,共5页Molecular Plant Breeding

基　　金：国家自然科学青年基金(31401432)资助

摘　　要：苎麻属于荨麻科苎麻属,是目前非常重要的纺织原料,有着广泛的开发前景。为了更好地研究苎麻基因组信息,拟通过流式细胞术测定苎麻基因组大小。本研究以水稻(Oryza sativa)日本晴为内标,湘苎3号为待测样品,分别分离水稻和苎麻叶片细胞的细胞核,PI染色,用流式细胞仪对水稻和苎麻样品的PI发射荧光强度进行测定。通过比较水稻和苎麻样品的荧光强度的倍数关系,计算得到苎麻基因组大小,并采用外标法和内标法同时进行测定和计算。以水稻日本晴为外标,苎麻3号为待测样品,测得苎麻基因组大小为332.5 Mb,而用内标法得到的苎麻基因组大小为300.5 Mb,相比之下内标法测定的变异系数更低,更为准确。苎麻基因组大小的测定为苎麻基因组文库的建立及其基因组相关研究提供了理论指导。Boehmeria nivea(ramie), which belongs to Urticaceae in Boehmeria, is an important textile material at present with a broad development prospects. In order to better study the genomic information of ramie, the genomic size of ramie was determined by flow cytometry. One of the ramie variants Xiangzhu 3 was used as the material and Oryza sativa Japonica as reference, and the nucleus of the leaf cells of both species were isolated and stained with propidium iodide(PI), then the PI emission fluorescence intensity was measured by flow cytometry.Ramie’s genome size was calculated according to the ratio of fluorescent peak values of Xiangzhu 3 to the rice reference subsequently. In addition, the external standard method and internal standard method are used for simultaneous determination and calculation. The estimated genome size of ramie was about 332.5 Mb by external standard method and about 330.5 Mb by internal standard method. The results showed that the internal method had smaller CV(coefficient of variation) value, which meant the result was more reliable. The determination of ramie’s genome size laid an important foundation for the construction of its genomic library and its genome related studies.

关 键 词：苎麻(Boehmeria nivea) 基因组大小 流式细胞术 水稻 

分 类 号：S563.1[农业科学—作物学]