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Development of a dense SNP-based linkage map of an apple rootstock progeny using the Malus Infinium whole genome genotyping array

机译:利用苹果属Infinium全基因组基因分型阵列,开发基于紧密SNP的苹果砧木后代连锁图谱

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Background A whole-genome genotyping array has previously been developed for Malus using SNP data from 28 Malus genotypes. This array offers the prospect of high throughput genotyping and linkage map development for any given Malus progeny. To test the applicability of the array for mapping in diverse Malus genotypes, we applied the array to the construction of a SNP-based linkage map of an apple rootstock progeny. Results Of the 7,867 Malus SNP markers on the array, 1,823 (23.2%) were heterozygous in one of the two parents of the progeny, 1,007 (12.8%) were heterozygous in both parental genotypes, whilst just 2.8% of the 921 Pyrus SNPs were heterozygous. A linkage map spanning 1,282.2?cM was produced comprising 2,272 SNP markers, 306 SSR markers and the S-locus. The length of the M432 linkage map was increased by 52.7?cM with the addition of the SNP markers, whilst marker density increased from 3.8?cM/marker to 0.5?cM/marker. Just three regions in excess of 10?cM remain where no markers were mapped. We compared the positions of the mapped SNP markers on the M432 map with their predicted positions on the ‘Golden Delicious’ genome sequence. A total of 311 markers (13.7% of all mapped markers) mapped to positions that conflicted with their predicted positions on the ‘Golden Delicious’ pseudo-chromosomes, indicating the presence of paralogous genomic regions or mis-assignments of genome sequence contigs during the assembly and anchoring of the genome sequence. Conclusions We incorporated data for the 2,272 SNP markers onto the map of the M432 progeny and have presented the most complete and saturated map of the full 17 linkage groups of M. pumila to date. The data were generated rapidly in a high-throughput semi-automated pipeline, permitting significant savings in time and cost over linkage map construction using microsatellites. The application of the array will permit linkage maps to be developed for QTL analyses in a cost-effective manner, and the identification of SNPs that have been assigned erroneous positions on the ‘Golden Delicious’ reference sequence will assist in the continued improvement of the genome sequence assembly for that variety.
机译:背景技术先前已经使用来自28种Malus基因型的SNP数据为Malus开发了全基因组基因分型阵列。该阵列为任何给定的Malus后代提供了高通量基因分型和连锁图谱开发的前景。为了测试该阵列在多种苹果属基因型中作图的适用性,我们将该阵列应用于构建基于SNP的苹果砧木后代的连锁图。结果在阵列上的7,867个Malus SNP标记中,两个后代中的一个亲本中有1,823个(23.2%)是杂合子,两个亲本基因型中有1,007个(12.8%)是杂合子,而在921个Pyrus SNP中只有2.8%是杂合子。杂合的。产生了一个跨越1,282.2?cM的连锁图,包含2,272个SNP标记,306个SSR标记和S位点。添加SNP标记后,M432连锁图的长度增加了52.7?cM,而标记密度从3.8?cM /标记增加到了0.5?cM /标记。只有三个超过10?cM的区域仍然没有标记被定位。我们将M432图谱上的SNP标记的位置与它们在“金冠”基因组序列中的预测位置进行了比较。共有311个标记(占所有映射标记的13.7%)映射到与其在'Golden Delicious'伪染色体上的预测位置冲突的位置,这表明在组装过程中存在同源基因组区域或基因组序列重叠群的错配和锚定基因组序列。结论我们将2272个SNP标记的数据整合到了M432后代的图谱中,并提出了迄今为止M. pumila的全部17个连接基团中最完整,最饱和的图谱。数据是在高通量半自动化管道中快速生成的,与使用微卫星构建链接地图相比,可以节省大量时间和成本。阵列的应用将允许以具有成本效益的方式开发连锁图谱用于QTL分析,并且鉴定在“ Golden Delicious”参考序列上分配了错误位置的SNP将有助于基因组的持续改进该品种的序列组装。

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