基于SLAF-seq技术的思茅松SNP位点开发

SNP Sites Developed by Specific Length Amplification Fragment Sequencing(SLAF-seq)in Pinus kesiya

为开发大量的思茅松SNP标记,本研究以思茅松两亲本和100个杂交的Fl代遗传群体为研究对象,利用SLAF-seq测序技术进行测序。选择HaeⅢ酶进行酶切,长度在414~464 bp的酶切片段定义为SLAF标签,然后将每个样本和标签序列比对,以获得不同样本的SNP标记信息。结果表明,建库双端比对效率在84.91%,酶切效率为95.52%,SLAF建库正常。共获得507394471个数据,测序平均Q30为92.25%,GC含量为40.23%,分布正常。共获得633086个SLAF标签,其中多态性SLAF标签有239790个,共有140485个标签成功编码,共开发获得11544个SNP标记。本研究结果可以对思茅松优良基因实行精细定位,为优良性状筛选、种质资源的鉴定和群体进化等相关研究提供新的依据。

In this study,the F generation genetic population of two parents and 100 crosses of Pinus kesiya var.langbianensis was sequenced using SLAF-seq sequencing technology.The HaeⅢenzyme was selected for digestion.The length of the enzyme section between 414~464 bp was defined as SLAF tag,and then each sample was compared with the tag sequence to get the SNP marker information of different samples.The results showed that the double-end comparison efficiency of library building was at 84.91%,the enzyme cutting efficiency was 95.52%,and the SLAF library building was normal.A total of 507394471 data were obtained,and the sequencing average Q30 was 92.25%and GC content was 40.23%,with normal distribution.A total of 633086 SLAF tags were obtained,of which 239790 were polymorphic SLAF tags,and a total of 140485 tags were successfully encoded.11544 SNP markers were developed and obtained by analyzing the obtained polymorphic SLAF tags.This study can further realize the fine localization of the excellent genes of Pinus kesiya var.langbianensis and provide a new basis for screening the excellent traits,identification of germplasm resources and population evolution and other related researches.