Topical delivery of dsRNA in two hemipteran species: Evaluation of RNAi specificity and non-target effects.

Double-stranded (ds) RNA-based technologies could provide novel and potential tool for pest management with efficiency and specificity of action. However, before applying this technique in the field, it is necessary to identify effective delivery methods and evaluate the non-target effects that may occur. In this article, we evaluated the effectiveness of dsRNA by topical delivery on a species of great agricultural interest, Halyomorpha halys. The specificity of action of the dsRNA was also investigated in Rhodnius prolixus, an insect phylogenetically close to H. halys. Of the three investigated genes (putative ATPase N2B, ATPase, serine/threonine-protein phosphatase PP1-ß catalytic subunit, PP1, and IAP repeat-containing protein 7-B-like, IAP), IAP and ATPase were able to induce higher mortality in H. halys nymphs compared to the control, with specific concentrations for each gene targeted. However, when the same RNAs were topically delivered to both R. prolixus 2nd and 3rd instar nymphs, no gene silencing and mortality were observed. For this reason, to assess dsRNA application-mediated non-target effects, we injected both H. halys and R. prolixus specific dsRNA in R. prolixus 5th instar nymphs. When the dsRNA targeting H. halys IAP was microinjected into R. prolixus 5th instar nymphs, no mortality was observed, suggesting a strong RNAi specificity. Together, these data suggest that the topical delivery could be suitable for the dsRNA to control H. halys population. Furthermore, its specificity of action would allow treatments towards single harmful species with limited non-target effects.

Effects of Acibenzolar-S-methyl on the Probing Behaviour and Mortality of Cacopsylla pyri on Pear Plants.

European pear psylla, Cacopsylla pyri, is one of the worst pests of pear in Europe. We investigated whether acibenzolar-S-methyl (ASM) application on pear plants might affect the behaviour in C. pyri. The elicitor was applied on pear potted plants, and after 48 h, we confirmed the ASM-mediated induction of several Pathogenesis-Related protein (PR) coding genes. At the same time, an in-depth analysis was performed on the probing behaviour of adults and nymphs of C. pyri on ASM-treated pear plants by the EPG-DC system, as well as the assessment of young nymphs' survival 7 days after the ASM application. The elicitor application weakly interfered with C. pyri nymphs probing behaviour and survival, while it did not affect adult stages. These data confirm previous observations obtained on C. pyricola and suggest that the elicitor does not represent a viable tool in the control of pear psylla species, especially if used alone, but it might be used in integrated management strategies focused on other plant pathogens such as Erwinia amylovora.

Monarda didyma Hydrolate Affects the Survival and the Behaviour of Drosophila suzukii.

Drosophila suzukii (Matsumara) is an herbivorous pest whose control in the field with conventional chemical is particularly difficult and has important drawbacks. Here, we investigated the insecticidal properties of hydrolate from Monarda didyma, scarlet beebalm, an aromatic herb in the Lamiaceae family. The identification of volatile organic compounds (VOCs) by CG-MS systems revealed that thymol (38%) and carvacrol (59%) were the most abundant VOCs in the hydrolate. M. didyma hydrolate did not show fumigant toxicity. Conversely, in contact assays, M. didyma hydrolate showed a LC50 of 5.03 µL mL-1, 48 h after the application on D. suzukii adults. Expression of detoxification genes increased in flies that survived the LC50 application. Furthermore, toxicity persisted for 7 days after the treatment in the survival evaluation. Artificial diet assays with 100 and 1000 µL mL-1 of M. didyma hydrolate resulted in a significant decrease in total food intake in both male and female D. suzukii adults. In addition, electropenetrography (EPG) showed that the D. suzukii females' feeding behaviour was altered in hydrolate-treated diets. The hydrolate also caused a significant reduction in the number of eggs laid in two different oviposition assays. Overall, our findings provide a new perspective for the improvement of D. suzukii control strategies through M. didyma hydrolate.