Kevei Zoltan, King Robert C, Mohareb Fady, Sergeant Martin J, Awan Sajjad Z, Thompson Andrew J
Cranfield Soil and AgriFood Institute, School of Energy, Environment and Agrifood, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, United Kingdom
Cranfield Soil and AgriFood Institute, School of Energy, Environment and Agrifood, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, United Kingdom Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom.
G3 (Bethesda). 2015 Mar 24;5(5):971-81. doi: 10.1534/g3.114.016121.
A recombinant in-bred line population derived from a cross between Solanum lycopersicum var. cerasiforme (E9) and S. pimpinellifolium (L5) has been used extensively to discover quantitative trait loci (QTL), including those that act via rootstock genotype, however, high-resolution single-nucleotide polymorphism genotyping data for this population are not yet publically available. Next-generation resequencing of parental lines allows the vast majority of polymorphisms to be characterized and used to progress from QTL to causative gene. We sequenced E9 and L5 genomes to 40- and 44-fold depth, respectively, and reads were mapped to the reference Heinz 1706 genome. In L5 there were three clear regions on chromosome 1, chromosome 4, and chromosome 8 with increased rates of polymorphism. Two other regions were highly polymorphic when we compared Heinz 1706 with both E9 and L5 on chromosome 1 and chromosome 10, suggesting that the reference sequence contains a divergent introgression in these locations. We also identified a region on chromosome 4 consistent with an introgression from S. pimpinellifolium into Heinz 1706. A large dataset of polymorphisms for the use in fine-mapping QTL in a specific tomato recombinant in-bred line population was created, including a high density of InDels validated as simple size-based polymerase chain reaction markers. By careful filtering and interpreting the SnpEff prediction tool, we have created a list of genes that are predicted to have highly perturbed protein functions in the E9 and L5 parental lines.
一个由樱桃番茄(E9)和醋栗番茄(L5)杂交产生的重组自交系群体已被广泛用于发现数量性状基因座(QTL),包括那些通过砧木基因型起作用的基因座,然而,该群体的高分辨率单核苷酸多态性基因分型数据尚未公开。对亲本系进行下一代重测序可以对绝大多数多态性进行表征,并用于从QTL推进到致病基因。我们分别对E9和L5基因组进行了40倍和44倍深度的测序,并将 reads 映射到参考海因茨1706基因组上。在L5中,1号染色体、4号染色体和8号染色体上有三个明显的多态性增加区域。当我们在1号染色体和10号染色体上比较海因茨1706与E9和L5时,另外两个区域具有高度多态性,这表明参考序列在这些位置包含一个不同的渐渗。我们还在4号染色体上鉴定出一个与醋栗番茄渗入海因茨1706一致的区域。创建了一个用于特定番茄重组自交系群体中精细定位QTL的多态性大数据集,包括大量经验证可作为基于简单大小的聚合酶链反应标记的插入缺失。通过仔细筛选和解释SnpEff预测工具,我们列出了在E9和L5亲本系中预测具有高度干扰蛋白功能的基因列表。