基于流式细胞术和K-mer分析的好好芭基因组大小估测  

Estimation of jojoba genome's size by flow cytometry and K-mer analysis

作　　者：马鹏举[1,2] 周佳熠 孙会改 马丹丹[1] 高飞 周宜君[1] 张根发[2] MA Pengju;ZHOU Jiayi;SUN Huigai;MA Dandan;GAO Fei;ZHOU Yijun;ZHANG Genfa(College of Life and Environmental Sciences,Minzu University of China,100081,Beijing,China;College of Life Sciences,Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University,100875,Beijing,China)

机构地区：[1]中央民族大学生命与环境科学学院,北京100081 [2]北京师范大学生命科学学院,抗性基因资源与分子发育北京市重点实验室,北京100875

出　　处：《北京师范大学学报（自然科学版）》2018年第6期733-737,共5页Journal of Beijing Normal University(Natural Science)

基　　金：国家自然科学基金资助项目(31670335,31770363,31470399);抗性基因资源与分子发育北京市重点实验室开放课题资助项目(201706);国家大学生创新性实验计划资助项目(GCCX2016110019);中央民族大学研究生自主科研基金资助项目(182115).

摘　　要：好好芭是一种重要的特种油料植物,具有极高的经济价值.由于好好芭基因组数据缺乏,导致其分子生物学研究进展相对缓慢.本研究以黄瓜和大豆作为内标,用流式细胞仪测定经碘化丙啶(PI)染色的内标细胞核和好好芭细胞核混合样品的PI荧光强度,通过比较好好芭与内标细胞DNA含量峰值的倍数关系,计算好好芭的基因组大小.同时使用基于生物信息学的K-mer分析来估测好好芭的基因组大小.采用流式细胞术测得好好芭基因组大小约为774 Mb.Kmer分析表明,当K值分别为17、19、21时,相应的DNA峰值出现在128、125和123,因此估测jojoba基因组大小为777～794Mb.K-mer分布曲线有明显的杂合峰,说明jojoba基因组杂合度较高.本研究完成了对好好芭基因组大小的估测,可为好好芭基因组学研究提供参考.Jojoba(Simmondsia chinensis)is an important oil plant with high economic value.Progress in molecular biology in jojoba research has been slow mainly due to lack of genomic sequence.In the present study,cucumber and soybean were used as internal standards,fluorescence intensity of mixed nucleus samples of internal standards and jojoba stained with propidium iodide were measured by flow cytometry.Genome size of jojoba was calculated by comparing the size of peak DNA content of jojoba with internal standard cells.K-mer analysis was used to estimate the size of jojoba genome.Jojoba genome size measured by flow cytometry was approximately 774 Mb.K-mer analyses showed that when K values were 17,19 and 21,corresponding DNA peaks appeared at 128,125 and 123,thus jojoba genome size should be between 777 Mb and 794 Mb.The K-mer distribution curve had a distinct heterozygous peak,indicating that genomic heterozygosity of jojoba was higher.This study estimated genome size of jojoba and provides a reference for further genomic research in jojoba.

关 键 词：好好芭 流式细胞术 K-mer分析 基因组大小 

分 类 号：S666.7[农业科学—果树学] R331[农业科学—园艺学]