| Summary: | <i>Thrips palmi</i> (Thysanoptera: Thripidae) is an important pest of vegetables, legumes, and ornamentals. In addition, it transmits several plant viruses. <i>T. palmi</i> genes associated with innate immunity, endocytosis-related pathways, and cuticular development are highly enriched in response to <i>Groundnut bud necrosis orthotospovirus</i> (GBNV, genus <i>Orthotospovirus</i>, family <i>Tospoviridae</i>) infection. As the previous transcriptomic study suggested the involvement of <i>T. palmi UHRF1BP1</i> and <i>PFAS</i> in GBNV infection, these two genes were targeted for silencing using antisense oligonucleotides (ASOs), and the effects on thrips’ fitness and virus acquisition were observed. Phosphorothioate modification of ASOs was carried out by replacing the nonbridging oxygen atom with a sulfur atom at the 3′ position to increase nuclease stability. The modified ASOs were delivered orally through an artificial diet. Exposure to ASOs reduced the target mRNA expression up to 2.70-fold optimally. Silencing of <i>T. palmi UHRF1BP1</i> and <i>PFAS</i> induced 93.33% mortality that further increased up to 100% with an increase in exposure. Silencing of <i>T. palmi UHRF1BP1</i> and <i>PFAS</i> also produced morphological deformities in the treated <i>T. palmi</i>. GBNV titer in <i>T. palmi</i> significantly declined post-exposure to ASOs. This is the first-ever report of silencing <i>T. palmi UHRF1BP1</i> and <i>PFAS</i> using modified ASO to induce mortality and impair virus transmission in <i>T. palmi</i>. <i>T. palmi UHRF1BP1</i> and <i>PFAS</i> would be novel genetic targets to manage thrips and restrict the spread of tospovirus.
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