Design of an eDNA sampling method for detection of an endophagous forest pest

Invasive wood-boring insects are a major economic and ecological concern worldwide as they impact native woody plant populations. These pest species are increasing in prevalence, with devastating impact, as global trade leads to higher rates of introduction and establishment. The emerald ash borer (Agrilus planipennis; EAB) is one such species, which has caused widespread damage across much of the United States and is now spreading across Europe. Non-indigenous woodborers such as EAB are difficult to detect at early stages of invasion, which is when management and eradication efforts are most effective and cost efficient. Environmental DNA (eDNA) surveys have demonstrated power in detecting invasive species when rare in the landscape due to their ability to detect trace amounts of DNA and identify to species. Here, we trialled a novel eDNA method for collecting environmental samples within host trees where invasive pest larvae are feeding, using EAB as a case study. We extracted tree cores approximately 1 cm in length using an increment hammer to assess detectability of eDNA from larvae feeding under the bark. In trees visibly infested with EAB, we observed a seasonal peak in EAB DNA detection probability (similar to 64%; towards the end of the growing season), indicating a potential impact of ash tree phenology or EAB phenology on detection. When we trialled the method in a site with ash trees of low or uncertain EAB abundance, we did not record positive EAB eDNA detections. This outcome may have resulted from differing EAB phenology at the northern latitude of this survey site or because larval galleries were less numerous causing EAB DNA to be scarcer within the tree. Our results, however, provide preliminary evidence that increment hammer tree cores can be used to detect eDNA of EAB and, perhaps, other wood-boring pests. Further work is needed to clarify false negative survey detections at ash trees showing little to no signs or symptoms of infestation, as well as investigating the deposition, transport and persistence dynamics of EAB eDNA within trees.