基于木薯全基因组重测序的InDel标记开发及应用

Development and Application of Cassava InDel Markers Based on Whole-genome Re-sequencing Datasets

本研究旨在开发稳定可靠、操作简便的木薯InDel分子标记,为木薯种质资源遗传多样性研究、品种鉴定等工作提供有力工具,为木薯育种发展提供保障。本研究基于22份木薯种质资源全基因组重测序分析结果随机挑选了64个InDel分子标记进行引物设计并利用琼脂糖凝胶电泳检测进行有效性验证,结果有20对引物具有多态性,多态性率为31.25%。随后将具有多态性的且扩增效果最优的18个InDel标记扩增条带数进行统计生成矩阵用于木薯种质资源遗传多样性研究,各标记的基因多样性指数于0.21~0.50之间,香农多样性指数于0.36~0.70之间。聚类结果分析将72份木薯材料在遗传相似系数0.62处划分为2大类群。且将中国的地方品种及选育品种大多划分至1#中的同一分支,其余薯肉颜色及内皮颜色相同的木薯材料大多都聚类到了一起。此外,本研究开发出的In Del分子标记IDC46具有特异性,可用于特定木薯材料鉴定。说明后期InDel引物有望成为木薯指纹图谱及分子身份证构建的有效工具,本研究结果也将对木薯育种工作后续开展具有一定的参考意义。

The aim of this study is to develop stable,reliable and simple molecular markers for cassava InDel,which will provide a powerful tool for exploring the genetic diversity of cassava germplasm resources,variety identification and other efforts,as well as providing assurance for the development of cassava breeding.Based on the genome-wide sequencing results of 22 cassava germplasm samples,64 InDel molecular markers were randomly selected for primer design and agarose gel electrophoresis validation.The results showed that 20 pairs of primers were polymorphic,with a polymorphism rate of 31.25 percent.The number of amplification bands of the 18 InDel markers with polymorphism and optimal amplification were then counted to study the genetic diversity of the cassava germplasm resource.The gene diversity index ranged from 0.21 to 0.50 for each marker and from 0.36 to 0.70 for Shannon.Based on the cluster analysis,72 cassava parents were divided into two groups with a genetic similarity factor of 0.62.Most of the landraces and cultivar were grouped in the same branch in 1#,and most of the other cassava materials of the same color of fresh root and color of root endothelial are grouped together.In addition,the InDel molecular marker IDC46 developed in this study is specific and can be used for the identification of specific cassava materials.In summary,the InDel primers were expected to be an effective tool for the construction of cassava fingerprints and molecular identity cards,and the results of this study will also have certain reference significance for the follow-up development of cassava breeding.