Londrina State University (UEL), Londrina, PR, Brazil.
Brazilian Agricultural Research Corporation - National Soybean Research Center (Embrapa Soja), Londrina, PR, Brazil.
Methods Mol Biol. 2022;2481:313-340. doi: 10.1007/978-1-0716-2237-7_18.
Soybean is one of the most valuable agricultural crops in the world. Besides, this legume is constantly attacked by a wide range of pathogens (fungi, bacteria, viruses, and nematodes) compromising yield and increasing production costs. One of the major disease management strategies is the genetic resistance provided by single genes and quantitative trait loci (QTL). Identifying the genomic regions underlying the resistance against these pathogens on soybean is one of the first steps performed by molecular breeders. In the past, genetic mapping studies have been widely used to discover these genomic regions. However, over the last decade, advances in next-generation sequencing technologies and their subsequent cost decreasing led to the development of cost-effective approaches to high-throughput genotyping. Thus, genome-wide association studies applying thousands of SNPs in large sets composed of diverse soybean accessions have been successfully done. In this chapter, a comprehensive review of the majority of GWAS for soybean diseases published since this approach was developed is provided. Important diseases caused by Heterodera glycines, Phytophthora sojae, and Sclerotinia sclerotiorum have been the focus of the several GWAS. However, other bacterial and fungi diseases also have been targets of GWAS. As such, this GWAS summary can serve as a guide for future studies of these diseases. The protocol begins by describing several considerations about the pathogens and bringing different procedures of molecular characterization of them. Advice to choose the best isolate/race to maximize the discovery of multiple R genes or to directly map an effective R gene is provided. A summary of protocols, methods, and tools to phenotyping the soybean panel is given to several diseases. We also give details of options of DNA extraction protocols and genotyping methods, and we describe parameters of SNP quality to soybean data. Websites and their online tools to obtain genotypic and phenotypic data for thousands of soybean accessions are highlighted. Finally, we report several tricks and tips in Subheading 4, especially related to composing the soybean panel as well as generating and analyzing the phenotype data. We hope this protocol will be helpful to achieve GWAS success in identifying resistance genes on soybean.
大豆是世界上最有价值的农作物之一。此外,这种豆类不断受到广泛的病原体(真菌、细菌、病毒和线虫)的攻击,从而降低产量并增加生产成本。主要的疾病管理策略之一是由单个基因和数量性状位点(QTL)提供的遗传抗性。鉴定大豆对这些病原体抗性的基因组区域是分子育种者首先要进行的步骤之一。过去,遗传图谱研究已被广泛用于发现这些基因组区域。然而,在过去的十年中,下一代测序技术的进步及其随后的成本降低导致了高通量基因分型的经济高效方法的发展。因此,应用数千个 SNP 在由不同大豆品系组成的大型集合中进行了全基因组关联研究。在本章中,提供了自该方法开发以来发表的大多数大豆疾病 GWAS 的综合回顾。由大豆疫霉、大豆疫霉和核盘菌引起的重要疾病一直是几项 GWAS 的重点。然而,其他细菌和真菌病也一直是 GWAS 的目标。因此,本 GWAS 综述可以作为未来这些疾病研究的指南。该方案首先描述了几种关于病原体的考虑因素,并带来了对它们进行分子特征描述的不同程序。提供了选择最佳分离株/种群的建议,以最大程度地发现多个 R 基因或直接映射有效的 R 基因。对几种疾病的大豆小组表型进行了方案、方法和工具的总结。我们还详细介绍了 DNA 提取方案和基因分型方法的选择,并描述了 SNP 质量参数对大豆数据的影响。强调了可用于获取数千个大豆品系的基因型和表型数据的网站及其在线工具。最后,在第 4 分节中报告了一些技巧和窍门,特别是与大豆小组的组成以及生成和分析表型数据有关的技巧和窍门。我们希望本方案将有助于在大豆中成功鉴定抗性基因的 GWAS。