In planta genetic transformation to produce CRISPRed high-oleic peanut

Compared to its normal-oleic counterpart, high-oleic peanut has better keeping quality and much more health benefits. Breeding high-oleic peanut through conventional means is a tedious process that typically takes several years. Genome editing, however, may shorten the duration. This study aimed to test the effectiveness of the node injection method coupled with CRISPR/Cas9 technology in inducing FAD2B mutations and high-oleic phenotype in peanut. Huayu 23, a popular normal-oleic runner type peanut cultivar having dysfunctional FAD2A and functional FAD2B , was transformed with CRISPR/Cas9 construct targeting FAD2B , resulting in two T 1 seeds with over 80% oleic acid and a 442 A insertion in FAD2B . The high-oleic phenotype in T 2 seeds was inheritable from the T 1 generation. As a genotype-independent, simple and easy method for peanut genetic transformation, node injection has great potential in functional analysis of genes and peanut varietal improvement. This method is of reference value to other seed plant species.