The spotted wing Drosophila (Drosophila suzukii) is an invasive and serious economic pest to small and stone fruits and its control is difficult. RNA interference (RNAi) or double-stranded RNA (dsRNA)-mediated gene silencing is rapidly becoming a widely used functional genomics tool in insects and holds great potential for insect pest control. This study investigates whether RNAi is functional in D. suzukii and whether oral delivery of dsRNA can elicit gene silencing and insecticidal activity. Firstly, microinjection of dsRNA targeting two essential genes (alpha COP and shrb) into the haemolymph of adult flies was performed, confirming that the RNAi system is functional and that gene silencing results in mortality. Secondly, dsRNA targeting alpha-COP and two extra essential genes, rpl13 and vha26, was mixed with artificial diet and fed to the larval and adult stages of D. suzukii. With naked dsRNA, no clear silencing and mortality were scored. However, combining dsRNA with a transfection reagent led to a significant increase in gene silencing and insect mortality. The best results were obtained with ds-Vha26. The results are discussed in relation to future optimization of the production, formulation, combinations and delivery of dsRNA
The South American tomato leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating pest in tomato crops. After having spread rapidly in the Mediterranean area since its first detection in 2006, it has recently become established in greenhouses in Western Europe. It was assumed that the low temperatures commonly associated with the winters in this area would prevent this exotic species from successful overwintering. However, the observed presence of T. absoluta at the start of the growing season in Belgium sparked a study on its potential to overwinter in Western Europe. In this study, the insect's cold hardiness and overwintering potential was assessed by determining: (1) the supercooling point (SCP) of larvae, pupae, and adults, (2) the lower lethal time (LT) for these stages at 0 and 5 A degrees C, and (3) the reproductive diapause incidence, as indicated by the reproductive performance of females reared at two temperatures (18 and 25 A degrees C) and two photoperiods [16:8 and 8:16 (L:D) h]. The mean SCP recorded for pupae (-16.7 A degrees C) was significantly higher than for adults (-17.8 A degrees C) and larvae (-18.2 A degrees C). Based on estimates of the LT adults demonstrated better cold hardiness than larvae and pupae at both 0 and 5 A degrees C with the median LT of adults averaging 17.9 and 27.2 days, respectively. No reproductive diapause was observed under the tested regimes. The results of this study indicate that T. absoluta is likely to successfully overwinter between two successive tomato crops in commercial greenhouses in Western Europe
RNA interference (RNAi) is a powerful tool to study functional genomics in insects and the potential of using RNAi to suppress crop pests has made outstanding progress. However, the delivery of dsRNA is a challenging step in the development of RNAi bioassays. In this study, we investigated the ability of engineered Flock House virus (FHV) to induce targeted gene suppression through RNAi under in vitro and in vivo condition. As proxy for fruit flies of agricultural importance, we worked with S2 cells as derived from Drosophila melanogaster embryos, and with adult stages of D. melanogaster. We found that the expression level for all of the targeted genes were reduced by more than 70% in both the in vitro and in vivo bioassays. Furthermore, the cell viability and median survival time bioassays demonstrated that the recombinant FHV expressing target gene sequences caused a significantly higher mortality (60–73% and 100%) than the wild type virus (24 and 71%), in both S2 cells and adult insects, respectively. This is the first report showing that a single stranded RNA insect virus such as FHV, can be engineered as an effective in vitro and in vivo RNAi delivery system. Since FHV infects many insect species, the described method could be exploited to improve the efficiency of dsRNA delivery for RNAi-related studies in both FHV susceptible insect cell lines and live insects that are recalcitrant to the uptake of naked dsRNA.
BACKGROUND: Previous studies have indicated the control potential of entomopathogenic nematodes (EPNs) against Tuta absoluta. Here, the potential of Steinernema feltiae, S. carpocapsae and Heterorhabditis bacteriophora is studied when applied against larvae of T. absoluta inside leaf mines in tomato leaf discs by means of an automated spray boom. RESULTS:The studied EPN species were effective against all four larval instars of T. absoluta but caused higher mortality in the later instars (e.g. fourth instar: 77.1-97.4% mortality) than in the first instars (36.8-60.0% mortality). Overall, S. feltiae and S. carpocapsae yielded better results than H. bacteriophora. Steinernema carpocapsae and H. bacteriophora performed better at 25 ∘ C (causing 55.3 and 97.4% mortality respectively) than at 18 ∘ C (causing 12.5 and 34.2% mortality respectively), whereas S. feltiae caused 100% mortality at both temperatures. Under optimal spraying conditions and with the use of Addit and Silwet L-77 adjuvants, a reduced dosage of 6.8 infective juveniles (IJs) cm −2 yielded equally good control as a recommended dosage of 27.3 IJs cm −2 .CONCLUSION: Under laboratory conditions, S. feltiae and S. carpocapsae showed good potential against the larvae of T. absoluta inside tomato leaf mines. Results need to be confirmed in greenhouse experiments.
M. pygmaeus can cause economic damage to tomato fruits at densities common in practice. An infection of the plants with PepMV enhances fruit symptoms significantly. Interacting plant defence responses are most likely the key to explaining this, although confirmation is required. © 2015 Society of Chemical Industry.
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