Identification of diagnostic KASP-SNP markers for routine breeding activities in yam (Dioscorea spp.)

Paterne A. Agre, Lindsay V. Clark, Ana Luisa Garcia-Oliveira, Rajaguru Bohar, Patrick Adebola, Robert Asiedu, Ryohei Terauchi, Asrat Asfaw
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Abstract

Maintaining genetic purity and true-to-type clone identification are important action steps in breeding programs. This study aimed to develop a universal set of kompetitive allele-specific polymerase chain reaction (KASP)-based single nucleotide polymorphism (SNP) markers for routine breeding activities. Ultra-low-density SNP markers were created using an initial set of 173,675 SNPs that were obtained from whole-genome resequencing of 333 diverse white Guinea yam (Dioscorea rotundata Poir) genotypes. From whole-genome resequencing data, 99 putative SNP markers were found and successfully converted to high-throughput KASP genotyping assays. The markers set was validated on 374 genotypes representing six yam species. Out of the 99 markers, 50 were highly polymorphic across the species and could distinguish different yam species and pedigree origins. The selected SNP markers classified the validation population based on the different yam species and identified potential duplicates within yam species. Through penalized analysis, the male parent of progenies involved in polycrosses was successfully predicted and validated. Our research was a trailblazer in validating KASP-based SNP assays for species identification, parental fingerprinting, and quality control (QC) and quality assurance (QA) in yam breeding programs.
为山药(薯蓣属)的常规育种活动鉴定诊断性 KASP-SNP 标记
保持基因纯度和真型克隆鉴定是育种计划的重要步骤。本研究旨在开发一套通用的基于竞争性等位基因特异性聚合酶链反应(KASP)的单核苷酸多态性(SNP)标记,用于常规育种活动。利用333个不同白几内亚山药(Dioscorea rotundata Poir)基因型的全基因组重测序获得的173,675个SNP初始集创建了超低密度SNP标记。从全基因组重测序数据中,发现了99个假定的SNP标记,并成功转化为高通量KASP基因分型分析。该标记集在6个山药品种的374个基因型上进行了验证。在99个标记中,有50个在物种间具有高度多态性,可以区分不同的山药品种和谱系来源。选择的SNP标记根据不同的山药物种对验证群体进行分类,并在山药物种中鉴定出潜在的重复。通过惩罚分析,成功地预测和验证了多交后代的父本。我们的研究在验证基于kasp的SNP分析用于物种鉴定、亲本指纹识别以及山药育种计划的质量控制(QC)和质量保证(QA)方面是开创性的。
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