基于SLAF-seq技术构建亚麻高密度遗传图谱  

High density genetic map of flax based on SLAF-seq technology

作　　者：高凤云[1,2] 斯钦巴特尔[2] 张辉[2] 贾霄云[2] 伊六喜[2] 周宇[2] 李强[2] 叶应福[3] 侯建华[1] 

机构地区：[1]内蒙古农业大学,内蒙古呼和浩特010019 [2]内蒙古农牧业科学院,内蒙古呼和浩特010031 [3]呼和浩特职业学院,内蒙古呼和浩特010051

出　　处：《中国油料作物学报》2017年第3期334-341,共8页Chinese Journal of Oil Crop Sciences

基　　金：内蒙古农牧业创新基金项目(2011CXJJM01-2);国家胡麻产业技术体系(CARS-17-GW-3);内蒙古自治区创新人才团队项目

摘　　要：为大量开发SNP用于构建亚麻高质量的连锁遗传图谱,以亚麻品系R43为母本,LH-89为父本,通过杂交构建F2群体,采用SLAF-seq技术,对两个亲本和F2群体100个个体进行高通量测序、SNP标记的开发及亚麻高密度遗传图谱的构建。对参考基因组进行电子酶切预测,最终确定使用HaeⅢ和RsaⅠ酶,酶切片段大小在314~414bp的序列定义为SLAF标签。其次,对基因组DNA进行酶切和SLAF文库构建。最后通过高通量测序获得测序数据,进行多态性SLAF的鉴定和SNP标记的发掘,采用High Map软件构建亚麻高密度遗传图谱。通过高通量测序,共获得196.29M reads,平均质量Q30为81.95%,平均GC含量为38.64%。通过生物信息学分析,共获得260 380个SNP标记,其中有4 547个SNP为高质量SNP标签,用于标记的定位和图谱的构建。共构建15个连锁群,总图距为2 632.94c M,标记间的平均距离为0.92c M。这是在亚麻中首次应用SNP标记构建的高密度遗传图谱,为亚麻的遗传研究和分子标记辅助育种奠定了基础。To develop high-throughput SNP markers for high quality map of flax（Linum usitatissimum L.）,100 individuals of 2 parents and their F2 population were detected by SLAF-seq technology. They were identified by high-throughput sequencing for SNP marker development and high-density genetic mapping. The 2 parents were varieties R43 and LH-89. After in silico digestion of reference genome,2 enzymes Hae Ⅲ and Rsa Ⅰ were used to digest the genome DNA. Restriction fragments of 314-414 bp were defined as SLAF. Polymorphic SLAFs were analysed and SNPs were detected according to data obtained by high-throughput sequencing. A high-density genetic map was constructed using High Map software. 196. 29 M reads were obtained,with average Q30 value of81. 95%,and average GC content 38. 64%. 260 380 SNPs were detected,of which 4 547 SNPs were qualified for genetic map construction. The final flax genetic map included 4 145 SNP markers on 15 linkage groups and was2 632. 94 c M in length with an average distance of 0. 96 c M between adjacent markers. This first high density genetic map in flax would be essential for genetic study and marker assistant breeding of flax.

关 键 词：亚麻 SLAF-seq技术 高通量测序 SNP标记 高密度遗传图谱 

分 类 号：S565.903[农业科学—作物学]