Characterization of five pigmentation genes as transgenic markers in Spodoptera frugiperda (Lepidoptera: Noctuidae)

The fall armyworm, Spodoptera frugiperda (J. E. Smith), has become one of the most damaging pests worldwide since its invasion of Africa, Asia and Oceania from 2016, threatening plants in 76 families including important crops. Genetics-based methods have proved to be an efficient way to control pests, especially invasive species, but many difficulties must be overcome to develop a transgenic insect strain, especially for a non-model species. Here we thus sought to identify a visible marker that would facilitate the distinction between genetically modified (GM) and non-transgenic insects, thereby simplifying mutation identification and facilitating the broader application of genome editing tools in non-model insects. Five genes (sfyellow-y, sfebony, sflaccase2, sfscarlet, and sfok) that are orthologs of well-studied genes in pigment metabolism were knocked out using the CRISPR/Cas9 system to identify candidate gene markers. Two genes, Sfebony and Sfscarlet, were identified responsible for body and compound eye coloration, respectively, in S. frugiperda, and could be potential visual markers for genetics-based pest management strategies.