Département de phytologie and Institut de biologie intégrative et des systèmes (IBIS), Université Laval, Quebec City, QC G1V 0A6, Canada.
Genome. 2018 Jun;61(6):449-456. doi: 10.1139/gen-2017-0236. Epub 2018 Apr 24.
Genotyping-by-sequencing (GBS) potentially offers a cost-effective alternative for SNP discovery and genotyping. Here, we report the exploration of GBS in tetraploid potato. Both ApeKI and PstI/MspI enzymes were used for library preparation on eight diverse potato genotypes. ApeKI yielded more markers than PstI/MspI but provided a lower read coverage per marker, resulting in more missing data and limiting effective genotyping to the tetraploid mode. We then assessed the accuracy of these SNPs by comparison with SolCAP data (5824 data points in diploid mode and 3243 data points in tetraploid mode) and found the match rates between genotype calls was 90.4% and 81.3%, respectively. Imputation of missing data did not prove very accurate because of incomplete haplotype discovery, suggesting caution in setting the allowance for missing data. To further assess the quality of GBS-derived data, a genome-wide association analysis was performed for flower color on 318 clones (with ApeKI). A strong association signal on chromosome 2 was obtained with the most significant SNP located in the middle of the dihydroflavonol 4-reductase (DFR) gene. We conclude that an appropriate choice of enzyme for GBS library preparation makes it possible to obtain high-quality SNPs in potato and will be helpful for marker-assisted genomics.
基于测序的基因分型(GBS)在 SNP 发现和基因分型方面具有成本效益高的潜力。本研究中,我们对四倍体马铃薯中的 GBS 进行了探索。在 8 个不同的马铃薯基因型上,我们分别使用 ApeKI 和 PstI/MspI 酶进行文库制备。ApeKI 产生的标记比 PstI/MspI 多,但每个标记的读取覆盖度较低,导致更多的数据缺失,并限制了有效基因分型到四倍体模式。然后,我们通过与 SolCAP 数据(二倍体模式下的 5824 个数据点和四倍体模式下的 3243 个数据点)进行比较来评估这些 SNP 的准确性,发现基因型调用之间的匹配率分别为 90.4%和 81.3%。由于不完全单倍型发现,缺失数据的插补并不十分准确,因此在设置缺失数据的允许值时要谨慎。为了进一步评估 GBS 衍生数据的质量,我们对 318 个克隆(使用 ApeKI)的花色进行了全基因组关联分析。在 DFR 基因中部的一个最强的 SNP 上获得了在染色体 2 上的强烈关联信号。我们得出结论,适当选择 GBS 文库制备的酶可以在马铃薯中获得高质量的 SNP,这将有助于基于标记的基因组学。