Laboratory sprayer for dsRNA application: Design and bioassay validation.

The shortage of commercially available and reliable laboratory spraying equipment for testing different preparations can be a major obstacle to achieve field-comparable results in the laboratory conditions. RNA interference is natural biological process which, when used for plant protection, can be designed method combining sustainability and minimal environmental side effects. Spraying of dsRNA is a field-relevant method that should ensure consistency and repeatability if conducted in laboratory. We built a portable spray device for laboratory use and tested its suitability for dsRNA application. For that, we carried out bioassay on three plant species with different leaf surface textures. DsRNA were detected in all samples 3 days post-treatment indicating its suitability for dsRNA delivery. We built a portable spray device for laboratory use and tested its suitability for dsRNA application. For that, we carried out:•Bioassay on three plant species with different leaf surface textures. DsRNA were detected in all samples 3 days post-treatment indicating its suitability for dsRNA delivery.

Spatiotemporal distancing of crops reduces pest pressure while maintaining conservation biocontrol in oilseed rape.

BACKGROUND: Agricultural landscapes provide resources for arthropod pests as well as their natural enemies. To develop integrated pest management (IPM) practices, it is important to understand how spatiotemporal location influences crop colonization and damage severity. We performed a 3-year (2016-2018) field experiment in winter oilseed rape (OSR, Brassica napus) fields in Estonia, where half of the fields were within 500 m of the location of the previous year's winter OSR field and half were outside this zone. We investigated how distance from the previous year's OSR crop influences the infestation and parasitism rates of two of its most important pests: the pollen beetle (Brassicogethes aeneus) and the cabbage seed weevil (Ceutorhynchus obstrictus). RESULTS: When the distance from the previous year's OSR crop was >500 m, we recorded significantly reduced pest pressure by both B. aeneus and C. obstrictus in the study fields. Biocontrol of both pests, provided by parasitic wasps, was high in each study year and commonly not affected by distance. Mean parasitism rates of B. aeneus were >31%, occasionally reaching >70%; for C. obstrictus, mean parasitism was >46%, reaching up to 79%, thereby providing effective biocontrol for both pest species. CONCLUSION: Spatiotemporal separation of OSR fields can reduce pest pressure without resulting in reduced parasitism of OSR pests. This supports a spatiotemporal field separation concept as an effective and sustainable technique for IPM in OSR. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Landscape complexity effects on Brassicogethes aeneus abundance and larval parasitism rate: a two-year field study.

Global biodiversity has suffered a decline primarily attributed to landscape simplification and intensified agricultural practices. Agricultural environments, characterized by homogeneity and frequent disturbances, are often suboptimal habitats for various insect species. While agricultural fields do favour pests, they generally fail to provide suitable habitats for natural enemies. The inclusion of diverse supporting habitats, such as semi-natural habitats, grassy and woody field margins etc. surrounding agricultural fields, play a crucial role in fostering effective biodiversity conservation. Moreover, determining the influence of different adjacent habitat types is essential in elucidating their influence on pest abundance and parasitism rates. Our two-year field study focused on assessing the abundance of Brassicogethes aeneus and its parasitism rate. The findings revealed that the adjacent habitat type did not significantly increase pest abundance and the parasitism rate of B. aeneus larvae consistently stayed over the threshold for effective biological control throughout the fields. This was attributed to the high proportion (35 and 38% in the 2 study years) of semi-natural habitats within most of the 1 km radius study areas. While our study did not identify any specific adjacent habitat type or habitat within a 1 km radius that directly impacted B. aeneus abundance, it emphasises the intricate interplay between the pests, parasitism and the surrounding environment because the interactive effect of distance from the crop edge and habitat type had a significant influence on B. aeneus infestation levels but not on parasitism. Decision tree analysis suggests that > 18% semi-natural habitat is needed to ensure sufficient levels of parasitism for effective biological control. A comprehensive understanding of habitats that influence not only B. aeneus but also other pests is critical for the successful implementation of IPM strategies and conservation initiatives within the agricultural sector.

Pollen beetle offspring is more parasitized under moderate nitrogen fertilization of oilseed rape due to more attractive volatile signal.

Biocontrol providing parasitoids can orientate according to volatile organic compounds (VOCs) of their host's plants, the emission of which is potentially dependent on the availability of soil nitrogen (N). This paper aimed at finding the optimal N fertilization rate for oilseed rape (Brassica napus L.) to favor parasitism of pollen beetles (Brassicogethes aeneus Fab. syn. Meligethes aeneus Fab.) in a controlled environment. Pollen beetles preferred to oviposit into buds of plants growing under higher N fertilization, whereas their parasitoids favored moderate N fertilization. As a part of induced defense, the proportion of volatile products of glucosinolate pathway in the total oilseed rape VOC emission blend was increased. Our results suggest that the natural biological control of pollen beetle herbivory is best supported by moderate N fertilization rates.

RNAi efficacy is enhanced by chronic dsRNA feeding in pollen beetle.

Double-stranded RNAs (dsRNAs) represent a promising class of biosafe insecticidal compounds. We examined the ability to induce RNA interference (RNAi) in the pollen beetle Brassicogethes aeneus via anther feeding, and compared short-term (3 d) to chronic (17 d) feeding of various concentrations of dsRNA targeting αCOP (dsαCOP). In short-term dsαCOP feeding, only the highest concentration resulted in significant reductions in B. aeneus survival; whereas in chronic dsαCOP feeding, all three concentrations resulted in significant mortality. Chronic dsαCOP feeding also resulted in significantly greater mortality compared to short-term feeding of equivalent dsαCOP concentrations. Our results have implications for the economics and development of dsRNA spray approaches for managing crop pests, in that multiple lower-concentration dsRNA spray treatments across crop growth stages may result in greater pest management efficacy, compared to single treatments using higher dsRNA concentrations. Furthermore, our results highlight the need for research into the development of RNAi cultivars for oilseed rape protection, given the enhanced RNAi efficacy resulting from chronic, compared to short-term, dsRNA feeding in B. aeneus.

First Evidence of Bud Feeding-Induced RNAi in a Crop Pest via Exogenous Application of dsRNA.

Spray-induced gene silencing (SIGS) is a potential strategy for agricultural pest management, whereby nucleotide sequence-specific double-stranded RNA (dsRNA) can be sprayed onto a crop; the desired effect being a consumption of dsRNA by the target pest, and subsequent gene silencing-induced mortality. Nucleotide sequence-specificity is the basis for dsRNA's perceived biosafety. A biosafe approach to pollen beetle (Brassicogethes aeneus) management in oilseed rape (Brassica napus) agroecosystems is needed. We examined the potential for SIGS in B. aeneus, via bud feeding, a field-relevant dsRNA exposure route. Oilseed rape buds were uniformly treated with dsRNA designed to target αCOP in B. aeneus. Our model control dsRNA (dsGFP) remained detectable on buds throughout the entire 3 d exposure period. When applied at 5 µg/µL, dsαCOP induced significant αCOP silencing 3 d after dietary exposure to buds treated with this dsαCOP concentration. We also observed a trend of increased αCOP silencing with increasing concentrations of dsαCOP at both 3 and 6 d. Furthermore, we observed a marginally significant and significant reduction in B. aeneus survival at 10 and 15 d, respectively. Our results suggest potential for developing a SIGS approach to B. aeneus management-though further experiments are needed to more fully understand this potential.

Effects of land use on infestation and parasitism rates of cabbage seed weevil in oilseed rape.

BACKGROUND: This study investigated how infestation rates of an important oilseed rape pest, the cabbage seed weevil (Ceutorhynchus obstrictus) and rates of parasitization by its parasitoids are affected by land use, up to 1000 m from 18 focal fields. RESULTS: The mean proportion of C. obstrictus-infested pods per plant was 8% (2-19.5%). Infestation rates were higher if the adjacent habitat was a herbaceous semi-natural habitat than if it was either another crop or a woody habitat. Infestation rates were positively related to the area of herbaceous semi-natural vegetation, permanent grassland and wheat (which followed oilseed rape in the crop rotation) at a spatial scale of at least 1 km. The mean parasitism rate of C. obstrictus larvae was 55% (8.3-87%), sufficient to provide efficient biocontrol. Parasitism rates were unrelated to adjacent habitats, however, they were positively related to the presence of herbaceous linear elements in the landscape and negatively related to permanent grasslands at a spatial scale of 200 m. CONCLUSION: Proximity of herbaceous elements increased both infestation rates and parasitism, while infestation was also related to landscape factors at larger distances. The findings provide an empirical basis for designing landscapes that suppress C. obstrictus, at both field and landscape scales. © 2018 Society of Chemical Industry.