Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes， including Δ9 desaturase， which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However， the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study， we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First， through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs， gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second... More
Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes， including Δ9 desaturase， which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However， the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study， we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First， through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs， gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second， using a sibling-crossing strategy， insects with mutant SlitDes11 (bearing a premature stop codon) were selected， and homozygous mutants were obtained in the G5 generation. Third， pheromone gland extracts of adult female homozygous SlitDes11 mutants were analyzed using Gas chromatography (GC). The results showed that titers of all three ester sex pheromone components; Z9， E11-14:Ac， Z9，E12-14:Ac， and Z9-14:Ac; were reduced by 62.40%， 78.50%， and 72.50%， respectively. This study provides the first direct evidence for the role of SlitDes11 in sex pheromone biosynthesis in S. litura， and indicates the gene could be as potential target to disrupt sexual communication in S. litura for developing a new pollution-free insecticide.
