Abstract
<title>Abstract</title> <p> RNA interference (RNAi) is a highly specific gene silencing mechanism activated by exogenous double-stranded RNA (dsRNA). dsRNA targeting a heat shock protein (dsHSP) specific to the highly invasive, tree-killing emerald ash borer (EAB) ( <italic>Agrilus planipennis</italic> ) has demonstrated efficacy against EAB with no phenotypic effects on several non-target insects. In soils, EAB dsHSP persistence and non-target research is underexplored. Therefore, we evaluated dsHSP for (1) persistence in soil, (2) effects on a model soil arthropod, <italic>Folsomia candida</italic> (Entomobryomorpha: Isotomidae), survival, body length, and reproduction, and (3) effects on abundance of a naturally occurring collembolan community. dsHSP treated soil was incubated for up to 7 days; total RNA was extracted at 4, 8, 24, 48, 72 hours and 7 days and then evaluated via RT-PCR. dsHSP was recovered at 4 and 8 hours post-treatment. In laboratory bioassays, <italic>F. candida</italic> exposed to soil treated with 200 µg dsHSP showed no effects on adult survival or body length after 28 days, but reproduction increased by 15%. In the field, ash trees received 750 mg dsHSP via foliar spray. Collembolans were sampled 14, 28, and 60 days post-treatment using pitfall traps and leaf litter vacuuming. Litter samples showed no treatment effect on abundance but pitfall samples near treated trees contained 11% fewer collembolans after 14 d and 59% more by 28 d. These results demonstrate short-term soil persistence with minimal adverse effects on non-target Collembola in laboratory and field settings, confirming RNAi specificity and providing insight into the exposure window for soil communities. </p>