CRISPR/Cas9-Mediated Siprolamin mutagenesis increase the content of functional amino acids in foxtail millet (Setaria italica)

Foxtail millet is a small diploid C4 crop that possesses superior nutritional properties. It provides a high content of essential amino acids, vitamins, and minerals. In this study, we used CRISPR/Cas9 to simultaneously edit two α-prolamin genes in foxtail millet. Analysis of the derived homozygous lines revealed a decrease in prolamin content. Surprisingly, the content of FAAs such as lysine, taurine, GABA, tryptophan, and sialic acid as well as glucose, fructose, sucrose, and maltose, was significantly increased. Additionally, peak viscosity, final viscosity, trough viscosity, and other parameters showed significant differences, suggesting that the food quality of the gene editing lines may have been altered. The editing lines grew normally, with only a slight decrease in 1000-grain weight compared to the wild type. Overall, we successfully created new germplasm with significantly increased FAAs content through gene editing, providing a valuable reference for the selection of functional foxtail millet varieties.