基于SLAF-seq技术的木薯SNP标记开发

Development of SNP markers in cassava(Manihot esculenta Crantz) based on SLAF-seq technology

开发大量木薯(Manihot esculenta)特异分子标记可促进木薯种质资源中优良基因的挖掘和利用。本研究以39份木薯种质资源为材料,利用特异性位点扩增片段测序(specific-locus amplified fragment sequencing,SLAF-seq)技术进行测序,结果显示:本次实验双端比对效率为90.83%,酶切效率为91.18%;通过测序获得304.30 Mb的读长数据,各样本读长范围在2 123 094~15 102 046之间,对照数据量最小,仅395 790个读长;样品测序质量值(Q_(30))在95.62%~96.60%之间,均在95%以上;GC含量在41.27%~44.96%之间,平均GC含量为42.89%,对照的GC含量为42.93%。本研究共开发出725 220个SLAF标签,样本的平均测序深度为20.98倍(×),多态性SLAF标签446 789个,共得到2 504 553个群体SNP标记。可见,SLAF-seq技术适用于木薯SNP位点的开发,其效率较高。

The development of a large number of specific molecular markers can promote the exploitation and utilization of the excellent genes in cassava（Manihot esculenta Crantz）germplasm resources.Specific-locus amplified fragment sequencing（SLAF-seq）technology was employed to sequence the genomes of 39 cassava varieties/lines in this study.Results show that paired-end alignment efficiency and cleavage efficiency were90.83%and 91.18%,respectively.304.30 Mb reads were obtained by sequencing,the reads of the samples varied from 2 123 094 to 15 102 046,and that of the control was the smallest,only 395 790.The sequencing quality value（Q（30））changed from 95.62%to 96.60%,all above 95%.The GC content of the samples changed from41.27%to 44.96%,the average value was 42.89%,and that of the control was 42.93%.Finally,725 220 SLAF labels were developed,among them 446 789 were polymorphic.The average sequencing depth of the samples was 20.98×,and 2 504 553 SNP markers were found.Obviously,SLAF-seq technology is suitable for developing more efficient SNP sites in cassava than ever.