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An integrated strategy for target SSR genotyping with toleration of nucleotide variations in the SSRs and flanking regions

文献类型： 外文期刊

作者： Huo, Yongxue 1 ; Zhao, Yikun 1 ; Xu, Liwen 1 ; Yi, Hongmei 1 ; Zhang, Yunlong 1 ; Jia, Xianqing 1 ; Zhao, Han 2 ; Zhao, J 1 ;

作者机构： 1.Beijing Acad Agr & Forest Sci BAAFS, Maize Res Ctr, Beijing Key Lab Maize DNA Fingerprinting & Mol Br, Beijing 100097, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Crop Germplasm & Biotechnol, Prov Key Lab Agrobiol, Nanjing 210014, Jiangsu, Peoples R China

关键词： SSR-GBS; Algorithm; Microsatellite; Sequence-based microsatellite genotyping; Genetic analysis

期刊名称：BMC BIOINFORMATICS （ 影响因子：3.169； 五年影响因子：3.629 ）

ISSN： 1471-2105

年卷期： 2021 年 22 卷 1 期

页码：

收录情况： SCI

摘要： Background: With the broad application of high-throughput sequencing and its reduced cost, simple sequence repeat (SSR) genotyping by sequencing (SSR-GBS) has been widely used for interpreting genetic data across different fields, including population genetic diversity and structure analysis, the construction of genetic maps, and the investigation of intraspecies relationships. The development of accurate and efficient typing strategies for SSR-GBS is urgently needed and several tools have been published. However, to date, no suitable accurate genotyping method can tolerate single nucleotide variations (SNVs) in SSRs and flanking regions. These SNVs may be caused by PCR and sequencing errors or SNPs among varieties, and they directly affect sequence alignment and genotyping accuracy. Results: Here, we report a new integrated strategy named the accurate microsatellite genotyping tool based on targeted sequencing (AMGT-TS) and provide a user-friendly web-based platform and command-line version of AMGT-TS. To handle SNVs in the SSRs or flanking regions, we developed a broad matching algorithm (BMA) that can quickly and accurately achieve SSR typing for ultradeep coverage and high-throughput analysis of loci with SNVs compatibility and grouping of typed reads for further indepth information mining. To evaluate this tool, we tested 21 randomly sampled loci in eight maize varieties, accompanied by experimental validation on actual and simulated sequencing data. Our evaluation showed that, compared to other tools, AMGT-TS presented extremely accurate typing results with single base resolution for both homozygous and heterozygous samples. Conclusion: This integrated strategy can achieve accurate SSR genotyping based on targeted sequencing, and it can tolerate single nucleotide variations in the SSRs and flanking regions. This method can be readily applied to divergent sequencing platforms and species and has excellent application prospects in genetic and population biology research. The web-based platform and command-line version of AMGT-TS are available at https://amgt-ts.plantdna.site:8445 and https://github.com/plantdna/amgt-ts, respectively.

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