Gene silencing in adult Popillia japonica through feeding of double‐stranded RNA (dsRNA) complexed with branched amphiphilic peptide capsules (BAPCs)
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ABSTRACT:
SUBMITTER: Carroll E
PROVIDER: S-EPMC10926504 | biostudies-literature | 2023 Jan
REPOSITORIES: biostudies-literature
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Project description:The Panel on Plant Health performed a pest categorisation of Popillia japonica (Coleoptera: Scarabaeidae) for the EU. P. japonica is a distinguishable species listed in Annex IAII of Council Directive 2000/29/EC. It is native to Japan but established in the USA in the early 20th century. It spreads from New Jersey to most US states east of the Mississippi, some to the west and north into Canada. P. japonica feeds on over 700 plant species. Adults attack foliage and fruit surfaces. They can cause serious injury to tree fruits and soft fruit, vegetable crops, ornamental herbaceous plants, shrubs, vines and trees. Larvae are root feeders regarded as serious pests of lawns and turf, vegetables and nursery stock. Adults emerge during the summer and can fly short distances on warm sunny days. The life cycle is usually completed in one year. In cooler regions, development takes two years. P. japonica occurs in the EU in the Azores (Portugal), Lombardy and Piedmont (Italy) where it is under official control. Adults are suspected of being able to spread on aircraft as hitchhikers, i.e. without host plants. Soil accompanying plants for planting provides a pathway for further introductions. Hosts are widely available within the EU. Climatic conditions across central and parts of southern EU are suitable for development in one year. Across parts of northern Europe development over two years is likely. Without control, impacts could be expected on a range of plants. Phytosanitary measures are available to reduce the likelihood of introduction of P. japonica. All criteria assessed by EFSA for consideration as a potential Union quarantine pest are met. Plants for planting are not necessarily the main means of spread so P. japonica does not satisfy all criteria necessary for it to be regarded as a Union regulated non-quarantine pest (RNQP).
| S-EPMC7009705 | biostudies-literature
Project description:Understanding cellular uptake mechanisms of nanoparticles with therapeutic potential has become critical in the field of drug delivery. Elucidation of cellular entry routes can aid in the dissection of the complex intracellular trafficking and potentially allow for the manipulation of nanoparticle fate after cellular delivery (i.e., avoid lysosomal degradation). Branched amphiphilic peptide capsules (BAPCs) are peptide nanoparticles that have been and are being explored as delivery systems for nucleic acids and other therapeutic molecules in vitro and in vivo. In the present study, we determined the cellular uptake routes of BAPCs with and without a magnetic nanobead core (BAPc-MNBs) in two cell lines: macrophages and intestinal epithelial cells. We also studied the influence of size and growth media composition in this cellular process. Substituting the water-filled core with magnetic nanobeads might provide the peptide bilayer nanocapsules with added functionalities, facilitating their use in bio/immunoassays, magnetic field guided drug delivery, and magnetofection among others. Results suggest that BAPc-MNBs are internalized into the cytosol using more than one endocytic pathway. Flow cytometry and analysis of reactive oxygen and nitrogen species (ROS/RNS) demonstrated that cell viability was minimally impacted by BAPc-MNBs. Cellular uptake pathways of peptide vesicles remain poorly understood, particularly with respect to endocytosis and intracellular trafficking. Outcomes from these studies provide a fundamental understanding of the cellular uptake of this peptide-based delivery system which will allow for strengthening of their delivery capabilities and expanding their applications both in vitro and in vivo.
| S-EPMC7962151 | biostudies-literature
Project description:Development of new and specific insect pest management methods is critical for overcoming pesticide resistance and collateral off-target killings. Gene silencing by feeding dsRNA to insects shows promise in this area. Here we described the use of a peptide nano-material, branched amphiphilic peptide capsules (BAPCs), that facilitates cellular uptake of dsRNA by insects through feeding. The insect diets included dsRNA with and without complexation with BAPCs. The selected insect species come from two different orders with different feeding mechanisms: Tribolium castaneum and Acyrthosiphon pisum. The gene transcripts tested (BiP and Armet) are part of the unfolded protein response (UPR) and suppressing their translation resulted in lethality. For Acyrthosiphon pisum, ingestion of BiP-dsRNA associated with BAPCs led to the premature death of the aphids (t1/2=4-5days) compared to ingestion of the same amounts of free BiP-dsRNA (t1/2=11-12days). Tribolium castaneum was effectively killed using a combination of BiP-dsRNA and Armet-dsRNA complexed with BAPCs; most dying as larvae or during eclosion (~75%). Feeding dsRNA alone resulted in fewer deaths (~30%). The results show that complexation of dsRNA with BAPCs enhanced the oral delivery of dsRNA over dsRNA alone.
| S-EPMC6984438 | biostudies-literature
Project description:BackgroundRNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphon pisum) genes can be achieved through direct injection of double-stranded RNA (dsRNA) or small-interfering RNAs (siRNA) into the pea aphid hemolymph or by feeding these insects on artificial diets containing the small RNAs.Methodology/principal findingsIn this study, we have developed the plant-mediated RNAi technology for aphids to allow for gene silencing in the aphid natural environment and minimize handling of these insects during experiments. The green peach aphid M. persicae was selected because it has a broad plant host range that includes the model plants Nicotiana benthamiana and Arabidopsis thaliana for which transgenic materials can relatively quickly be generated. We targeted M. persicae Rack1, which is predominantly expressed in the gut, and M. persicae C002 (MpC002), which is predominantly expressed in the salivary glands. The aphids were fed on N. benthamiana leaf disks transiently producing dsRNA corresponding to these genes and on A. thaliana plants stably producing the dsRNAs. MpC002 and Rack-1 expression were knocked down by up to 60% on transgenic N. benthamiana and A. thaliana. Moreover, silenced M. persicae produced less progeny consistent with these genes having essential functions.Conclusions/significanceSimilar levels of gene silencing were achieved in our plant-mediated RNAi approach and published silencing methods for aphids. Furthermore, the N. benthamiana leaf disk assay can be developed into a screen to assess which genes are essential for aphid survival on plants. Our results also demonstrate the feasibility of the plant-mediated RNAi approach for aphid control.
| S-EPMC3187792 | biostudies-literature
Project description:Popilia japonica principal haplotype assembly genome sequencing
| PRJNA1120502 | ENA
Project description:Japanese beetle (Popillia japonica) genome sequencing and assembly
| PRJNA514809 | ENA
Project description:Popilia japonica alternate haplotype assembly genome sequencing
| PRJNA1120503 | ENA
Project description:Popillia japonica Genome sequencing and assembly
| PRJNA1172713 | ENA