Description
Soybean (Glycine max) is a widely cultivated crop, valued for its high protein and oil content. There is growing interest in enhancing soybean fatty acid composition for nutritional and commercial purposes. The nutritional and industrial value of soybean oil is dependent on fatty acid biosynthesis; therefore, efforts to refine its oil profiles have focused on key biosynthetic pathways. Fatty acid biosynthesis is regulated by the acyl-ACP thioesterase (FAT) enzyme family, which controls the chain length and saturation of fatty acids. Numerous members of the FAT family have been characterized; however, several remain unexplored. This study builds on the previous characterization of the FAT gene family using a reverse genetics approach. More specifically, the characterization of FATB-3A, FATB-3B, FATB-4A, FATB-5A, and FATB-5B genes. A TILLING-by-Sequencing+ strategy was employed to identify mutations in target FAT genes within an EMS-mutagenized population. Mutations were validated using Sanger sequencing, and changes in fatty acid composition were assessed. Protein homology modeling and subcellular localization of all FAT members were carried out. Lastly, RNA-seq analysis was conducted to observe tissue-specific expression patterns. These comprehensive analyses enhance our understanding of FAT gene function in soybean oil structure and support the development of varieties with specific oil profiles for health and industrial use. The developed soybeans may positively influence the development of high-oleic-acid lines to improve the nutritional value of soybeans without potential development tradeoffs.