Improving digestibility of sorghum proteins by CRISPR/Cas9 ‐based genome editing

In this study, we used CRISPR/Cas9 gene editing to target theK2G gene to create new sorghum lines with reduced levels of kafirin. A guide RNA (sgRNA) was designed to introduce mutations in the CDS region that encodes the endoplasmic reticulum signal peptide of γ-kafirin. The pK2GsgRNA/Cas9 vector was transformed into sorghum using the pollen-mediated transformation method. Sequencing of the transformants showed that three out of 24 transgenic plants contain genetic mutations in the targeted region. Compared with the wildtype, the γ-kafirin contents of the mutant plants decreased by 12.75%–19.22%, and the protein digestibility of the mutant kernels increased by 26.91%–74.31% in raw flour. Although the grain weights remained comparable to those of the wildtype, the growth of the mutant plants was more vigorous as the muta nt shoots grew taller and thicker compared with those of the wildtype. Our work advances the ability to improve the digestibility of an important crop. The resulting quality improvements can also be rapidly deployed for breeding and generation of transgene-free, improved cultivars of sorghum, a majo r crop worldwide.
Source: Food and Energy Security - Category: Food Science Authors: Tags: ORIGINAL ARTICLE Source Type: research