The Kunitz trypsin inhibitor (KTI) gene family encompasses a category of trypsin inhibitors, and the KTI proteins are important components of the 2S storage protein fraction in soybeans. In this study, fifty members of the GmKTI family were identified in the soybean genome, and their physicochemical properties, domain compositions, phylogenetic relationships, gene structures, and expression patterns were comprehensively analyzed to explore their impact on soybean seed protein content. The results revealed significant gene expansion within the GmKTI family in soybean. The gene structures and conserved motifs of GmKTI members exhibited both regularity and diversity, with distinct expression patterns across different soybean tissues. Haplotype analysis identified 7 GmKTI genes significantly associated with seed storage protein content, and the combination of superior haplotypes was found to enhance seed storage protein content. This is crucial for the improvement of soybean varieties and the enhancement of storage protein content. Additionally, the GmKTI family demonstrated evolutionary conservation, with its functions likely linked to light induction, biotic stress, and growth development. This study characterizes the structure, expression, genomic haplotypes, and molecular features of the soybean KTI domain for the first time, providing a foundation for functional analyses of the GmKTI domain in soybean and other plants.

The dimensions of organs such as flowers, leaves, and seeds are governed by processes of cellular proliferation and expansion. In soybeans, the dimensions of these organs exhibit a strong correlation with crop yield, quality, and other phenotypic traits. Nevertheless, there exists a scarcity of research concerning the regulatory genes influencing flower size, particularly within the soybean species. In this study, 309 samples of 3 soybean types (123 cultivar, 90 landrace, and 96 wild) were re-sequenced. The microscopic phenotype of soybean flower organs was photographed using a three-eye microscope, and the phenotypic data were extracted by means of computer vision. Pearson correlation analysis was employed to assess the relationship between petal and seed phenotypes, revealing a strong correlation between the sizes of these two organs. Through GWASs, SNP loci significantly associated with flower organ size were identified. Subsequently, haplotype analysis was conducted to screen for upstream and downstream genes of these loci, thereby identifying potential candidate genes. In total, 77 significant SNPs associated with vexil petals, 562 significant SNPs associated with wing petals, and 34 significant SNPs associated with keel petals were found. Candidate genes were screened by candidate sites, and haplotype analysis was performed on the candidate genes. Finally, the present investigation yielded 25 and 10 genes of notable significance through haplotype analysis in the vexil and wing regions, respectively. Notably, Glyma.07G234200, previously documented for its high expression across various plant organs, including flowers, pods, leaves, roots, and seeds, was among these identified genes. The research contributes novel insights to soybean breeding endeavors, particularly in the exploration of genes governing organ development, the selection of field materials, and the enhancement of crop yield. It played a role in the process of material selection during the growth period and further accelerated the process of soybean breeding material selection.