摘要： 高粱在粮食安全、人类健康和生物能源的开发上具有非常重要的作用，其被认为是二倍体模式化作物和多倍体能源作物甘蔗和芒草类的参考植物，开发高粱多态性分子标记对促进分子遗传学发展及高粱分子标记辅助育种意义重大。以甜高粱与粒用高粱杂交的F₂遗传群体作为研究材料，对其全基因组DNA进行酶切，然后用高通量测序法开发SLAF（Specific-locus amplified fragment sequencing）标签，并经过分析筛选多态性SNP标记。试验过程中测序的reads数达到43 528 021个，母本2 598 472个，父本3 134 524个，子代的reads数为205 083~454 258个，平均为290 732.5个。通过原始数据的评估、聚类和纠错开发 的SLAF标签数，父母本分别为44 895，42 100个。测序深度分别为19.78和16.22，F₂ 群体每个个体的 SLAF 标签为 26 737~39 291个，平均为 33 445.06个，测序深度2.24~3.72，平均为2.79。通过对聚类群中高深度片段的潜在基因型分析，得到了 6 353 个多态性 SNP 标记，其中 5 829 个标记成功分型，占多态性 SNP 标记的91.75%，剔除不适宜作图的标记，剩余有效的 SNP 标记 2 246 个，占有效标记百分比100%，F₂个体的各样品完整度平均为94.99%。所以，对基因组DNA进行酶切和高通量测序，从庞大的reads中获得SLAF标签是得到全基因组SNP标记的有效途径。为高粱高密度图谱构建和QTL定位研究，以及分子标记辅助育种奠定了基础。

关键词: 高粱, SLAF标签, 高通量测序, 开发, SNP标记

Abstract: Sorghum is a global crop for food security,human health and bio-energy development. It is considered one of the model crops for diploid cereal species and a reference crop for polyploid species like sugarcane and Miscanthum. It is important to develop molecular markers for genetics research and marker assisted selection in sorghum breeding. The parental lines,a sweet sorghum variety and a grain sorghum variety,and their F₂ progenies were employed to develop Specific-Locus Amplified Fragment Sequencing(SLAF) markers,from which a large amount of SNP markers were obtained by using a high-throughput sequencing technology. A total of 43 528 021 SLAF-seq reads were generated,including 2 598 472 reads from the maternal variety,and 3 134 524 reads from the paternal variety,and 205 083 to 454 258 reads with an average of 290 732.5 from each F₂ individual. High quality SLAF tags generated form the maternal and the paternal varieties were 44 895 and 42 100,with sequencing depth of 19.78 and 16.22-fold,respectively. High quality SLAF tags from the F₂ progenies ranged from 26 737 to 39 291(average 33 445.06) with a sequencing depth of 2.24 to 3.72-fold(average 2.79). A total of 6 353 polymorphic SNP markers were obtained,in which 5 829(91.75%) were successfully genotyped and 2 246 high quality markers have been obtained with 100% efficient and an average of 94.99% integrity. Results showed that it was a useful way to obtain SNP markers covering whole genome through high-throughput sequencing technology. This study has laid a foundation for linkage map construction,QTL identification and marker assisted breeding in sorghum.

Key words: Sorghum, SLAF-tag, High-throughput sequencing, Development, SNP Marker

中图分类号: 

• S514.03