Fluorescent-Based Sex-Separation Technique in Major Invasive Crop Pest, <i>Drosophila suzukii</i>

Insect population biocontrol methods, such as the sterile insect technique (SIT), offer promising alternatives to traditional pesticide-based approaches for managing pest populations. Effective use of these strategies requires scalable sex separation techniques, which are currently nonexistent for Drosophila suzukii , a prominent crop pest species. SEPARATOR (Sexing Element Produced by Alternative RNA-splicing of A Transgenic Observable Reporter) is a fluorescence-based sex-sorting technology previously developed for other pest species that enables the identification of females and males based on sex-specific fluorescence. In this study, we investigate the applicability of SEPARATOR to D. suzukii by engineering several strains encoding SEPARATOR constructs that facilitate efficient sex selection in early larval stages. Our findings demonstrate the feasibility of using SEPARATOR in D. suzukii by incorporating transformer introns from three different species: D. suzukii , D. melanogaster , and Ceratitis capitata . This study provides an initial basis for further optimization of SEPARATOR in D. suzukii and indicates that it could be a helpful tool for SIT-based biocontrol strategies.