<p>Sorghum grain mold is a devastating fungal complex whose genetic dissection is frequently hindered by polygenic architecture and inconsistent marker signals. To resolve this complexity, we developed a unified GWAS framework that integrates standard single-nucleotide polymorphisms (SNPs) with a local haplotype-window marker layer (“Haplo-<i>k</i>mer;” <i>k</i>-SNP windows encoded from the GBS SNP matrix, not read-derived <i>k</i>-mer counts), prioritizing genomic intervals where evidence converges across multiple traits and methods. Leveraging the Sorghum Association Panel (SAP) and six distinct phenotype indices, we identified a convergent core interval under an operational multi-trait, multi-method convergence framework (<i>p</i> <InlineEquation ID="IEq1"> <EquationSource Format="TEX">\(\le 5\)</EquationSource> <EquationSource Format="MATHML"><math> <mrow> <mo>≤</mo> <mn>5</mn> </mrow> </math></EquationSource> </InlineEquation>× 10<sup>–4</sup>) on Chr5 (60.25–60.50&#xa0;Mb). This interval was supported by multi-trait convergence and rank-preserving permutation analyses, representing a convergent hub supported by three distinct resistance-related traits and reinforced by haplotype-based association signals (<InlineEquation ID="IEq2"> <EquationSource Format="TEX">\(\Sigma -{\text{log}}_{10}\left(p\right)=32.43\)</EquationSource> <EquationSource Format="MATHML"><math> <mrow> <mi mathvariant="normal">Σ</mi> <mo>-</mo> <msub> <mtext>log</mtext> <mn>10</mn> </msub> <mfenced close=")" open="("> <mi>p</mi> </mfenced> <mo>=</mo> <mn>32.43</mn> </mrow> </math></EquationSource> </InlineEquation>). Decomposition of the signal revealed that while SNPs captured isolated hits for <i>Alternaria</i> (A) and <i>Curvularia</i> (C) indices, the mixed-inoculum (M) index was supported by both SNP and Haplo-<i>k</i>mer markers, alongside strong contributions from A and C indices. We validated the biological relevance of this locus through unsupervised haplotype clustering of 54 regional SNPs, which successfully stratified sorghum lines into distinct phenotypic classes (Kruskal–Wallis <i>p</i> = 0.0029). Furthermore, a genome-wide placebo test demonstrated that this haplotype–phenotype stratification shows moderate specificity against the random genomic background (empirical <i>p</i> = 0.165). Collectively, these findings delineate a robust candidate interval for marker-assisted tracking on Chr5 and demonstrate how haplotype-aware strategies can recover fragmented association evidence in a polygenic pathosystem.</p>

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A haplotype-layered GWAS identifies a multi-trait grain mold resistance hub on sorghum chromosome 5

  • Ezekiel Ahn,
  • Sookyung Oh,
  • Louis K. Prom,
  • Seunghyun Lim,
  • Lyndel W. Meinhardt,
  • Clint Magill

摘要

Sorghum grain mold is a devastating fungal complex whose genetic dissection is frequently hindered by polygenic architecture and inconsistent marker signals. To resolve this complexity, we developed a unified GWAS framework that integrates standard single-nucleotide polymorphisms (SNPs) with a local haplotype-window marker layer (“Haplo-kmer;” k-SNP windows encoded from the GBS SNP matrix, not read-derived k-mer counts), prioritizing genomic intervals where evidence converges across multiple traits and methods. Leveraging the Sorghum Association Panel (SAP) and six distinct phenotype indices, we identified a convergent core interval under an operational multi-trait, multi-method convergence framework (p \(\le 5\) 5 × 10–4) on Chr5 (60.25–60.50 Mb). This interval was supported by multi-trait convergence and rank-preserving permutation analyses, representing a convergent hub supported by three distinct resistance-related traits and reinforced by haplotype-based association signals ( \(\Sigma -{\text{log}}_{10}\left(p\right)=32.43\) Σ - log 10 p = 32.43 ). Decomposition of the signal revealed that while SNPs captured isolated hits for Alternaria (A) and Curvularia (C) indices, the mixed-inoculum (M) index was supported by both SNP and Haplo-kmer markers, alongside strong contributions from A and C indices. We validated the biological relevance of this locus through unsupervised haplotype clustering of 54 regional SNPs, which successfully stratified sorghum lines into distinct phenotypic classes (Kruskal–Wallis p = 0.0029). Furthermore, a genome-wide placebo test demonstrated that this haplotype–phenotype stratification shows moderate specificity against the random genomic background (empirical p = 0.165). Collectively, these findings delineate a robust candidate interval for marker-assisted tracking on Chr5 and demonstrate how haplotype-aware strategies can recover fragmented association evidence in a polygenic pathosystem.