Abstract
Reliable identification of genomic regions controlling complex agronomic traits across variable growing seasons remains a major challenge in soybean genetics and breeding. Here, a diverse panel of 252 soybean accessions was evaluated over six consecutive growing seasons (2018–2023) for flowering time, maturity, plant height, number of seeds per plant, seed yield per plant, and thousand-seed weight. Whole-genome resequencing and variant filtering yielded 2,019,772 high-quality SNPs, and association signals were evaluated using Inclusive Integrative Input Multiple-locus Random-SNP-effect Mixed Linear Model (IIIVmrMLM), Bayesian-information and Linkage-disequilibrium Iteratively Nested Keyway (BLINK), and Multi-Locus Mixed Model (MLMM) together with linkage disequilibrium (LD)-based locus consolidation. Cross-model prioritization retained 21 high-confidence loci supported by all three GWAS models and distributed across 10 chromosomes. Among the identified loci, 18 overlapped or co-localized with previously reported SoyBase genes and QTLs, whereas three (q.VER2.13-1, q.YP.01-1, and q.TSW.15-1) showed no positional overlap with known genes and QTLs and were therefore considered presumably novel. These three loci were associated with flowering time, yield per plant, and thousand-seed weight, accounting for 1.60%, 2.13%, and 5.53% of phenotypic variation, respectively. Ten loci co-localized with genomic regions containing established soybean regulators, including E2, E3, GmDt2, and POWR1, support the biological plausibility of the association results. Integration of genomic position, functional annotation, and tissue-expression evidence prioritized 112 candidate genes across 18 loci. These findings provide a focused set of genomic loci and candidate genes for independent validation and further investigation of the genetic basis of soybean adaptation and yield formation under variable continental growing conditions.