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Pinpointing genes underlying the quantitative trait loci for root-knot nematode resistance in palaeopolyploid soybean by whole genome resequencing

机译:通过全基因组重排来定位古多倍体大豆根结线虫抗性的数量性状基因位点的基础基因

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摘要

The objective of this study was to use next-generation sequencing technologies to dissect quantitative trait loci (QTL) for southern root-knot nematode (RKN) resistance into individual genes in soybean. Two hundred forty-six recombinant inbred lines (RIL) derived from a cross between Magellan (susceptible) and PI 438489B (resistant) were evaluated for RKN resistance in a greenhouse and sequenced at an average of 0.19× depth. A sequence analysis pipeline was developed to identify and validate single-nucleotide polymorphisms (SNPs), infer the parental source of each SNP allele, and genotype the RIL population. Based on 109,273 phased SNPs, recombination events in RILs were identified, and a total of 3,509 bins and 3,489 recombination intervals were defined. About 50.8% of bins contain 1 to 10 genes. A linkage map was subsequently constructed by using bins as molecular markers. Three QTL for RKN resistance were identified. Of these, one major QTL was mapped to bin 10 of chromosome 10, which is 29.7 kb in size and harbors three true genes and two pseudogenes. Based on sequence variations and gene-expression analysis, the candidate genes underlying the major QTL for RKN resistance were pinpointed. They are Glyma10g02150 and Glyma10g02160, encoding a pectin methylesterase inhibitor and a pectin methylesterase inhibitor -pectin methylesterase, respectively. This QTL mapping approach not only combines SNP discovery, SNP validation, and genotyping, but also solves the issues caused by genome duplication and repetitive sequences. Hence, it can be widely used in crops with a reference genome to enhance QTL mapping accuracy.
机译:这项研究的目的是使用下一代测序技术将对南方根结线虫(RKN)抗性的定量性状基因座(QTL)分解为大豆中的单个基因。对来自麦哲伦(易感)和PI 438489B(抗性)之间的杂交的246个重组自交系(RIL)进行了温室大棚RKN抗性评估,平均测序深度为0.19倍。开发了序列分析流程以鉴定和验证单核苷酸多态性(SNP),推断每个SNP等位基因的亲本来源,并对RIL群体进行基因分型。基于109,273个分阶段的SNP,确定了RIL中的重组事件,共定义了3,509个bin和3,489个重组间隔。大约50.8%的垃圾箱包含1到10个基因。随后通过使用箱作为分子标记构建连锁图。确定了三个针对RKN抗性的QTL。其中,一个主要的QTL被定位到10号染色体的10号位,大小为29.7 kb,具有三个真实基因和两个假基因。基于序列变异和基因表达分析,确定了针对RKN抗性的主要QTL的候选基因。它们分别是Glyma10g02150和Glyma10g02160,分别编码果胶甲基酯酶抑制剂和果胶甲基酯酶抑制剂-果胶甲基酯酶。这种QTL映射方法不仅结合了SNP发现,SNP验证和基因分型,而且解决了由基因组重复和重复序列引起的问题。因此,它可以广泛用于具有参考基因组的农作物中,以提高QTL作图的准确性。

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