不同生态类型菘蓝SLAF-seq分子标记

SLAF-seq molecular markers in different ecological types of Isatis tinctoria

目的探究不同生态类型菘蓝Isatis tinctoria表型差异和特异位点扩增片段测序(specific-locus amplified fragment sequencing,SLAF-seq)分子标记。方法以25个不同生态类型菘蓝为研究对象,利用SLAF-seq技术开发SLAF标签,再结合菘蓝叶片叶表观特征的田间观测,分析不同生态类型菘蓝植株表观差异。结果测序后各样品所获得的读长总量各不相同,测序质量值Q30为95.66%~96.60%,GC比例为38.13%~41.37%。共开发535105个菘蓝SLAF标签,多态性SLAF标签有34412个,共得到63002个群体单核苷酸多态性(single nucleotide polymorphism,SNP)标记。测序分析结果与叶表观特征调查结果相结合,可以将25个不同生态类型菘蓝分为5类,分别为倒卵圆形无缺刻皱缩叶、倒卵圆形缺刻皱缩叶、倒卵圆形无缺刻平坦叶、倒卵圆形缺刻平坦叶、披针形无缺刻平坦叶。结论为后期进一步研究不同类型菘蓝的基因来源和遗传进化提供参考,为菘蓝种质分类、品种鉴定和分子辅助育种提供借鉴。

Objective To explore the phenotypic differences and specific-locus amplified fragment sequencing(SLAF-seq)molecular markers of Isatis indigogenes from different ecological types.Methods Using twenty-five different ecological types of Isatis indigotica as research objects,SLAF labels were developed by SLAF-seq technique,and the apparent differences of Isatis indigotica plants were analyzed by field observation of leaf apparent characteristics.Results After sequencing,the total read length of each sample was different,the sequencing quality value Q30 was 95.66%—96.60%,and the GC ratio was 38.13%—41.37%.A total of 535105 Isatis indigotica SLAF labels and 34412 polymorphic SLAF labels were developed,resulting in 63002 single nucleotide polymorphism(SNP)markers.Combining the results of sequencing analysis with the investigation of leaf apparent characteristics,the 25 different ecological types of Isatis indigotica could be divided into five categories,namely,obovate without engraved wrinkled leaves,obovate with engraved wrinkled leaves,obovate without engraved flat leaves,obovate with engraved flat leaves,lanceolate without engraved flat leaves.Conclusion It provides reference for further study on gene origin and genetic evolution of different types of Isatis indigotica,and provides reference for germplasm classification,variety identification and molecular assisted breeding of Isatis indigotica.