RNA interference protocols for gene silencing in the spittlebug Philaenus spumarius, vector of Xylella fastidiosa.
Double stranded RNA
Microinjection
Plant-mediated feeding
dsRNA delivery strategies
smallRNA sequencing
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
28 10 2024
28 10 2024
Historique:
received:
29
05
2024
accepted:
23
09
2024
medline:
29
10
2024
pubmed:
29
10
2024
entrez:
29
10
2024
Statut:
epublish
Résumé
RNA interference (RNAi) is double stranded RNA (dsRNA)-based gene silencing mechanism. Exogenous dsRNAs application to crops has raised as a powerful tool to control agricultural pests. In particular, several sap-feeder are important plant pathogens vectors, such as Philaenus spumarius, known as main vector of Xylella fastidiosa (Xf), causal agent of olive quick decline syndrome (OQDS) in southern Italy. Here, dsATP synthase beta (dsATP), dsLaccase (dsLacc) and dsGreen Fluorescent Protein (dsGFP) as control, were provided to spittlebug adults by microinjection or to nymphs fed on dsRNA-treated plant shoots. Treated insects were collected at different time points to monitor silencing efficiency over time, describing significant reduction of transcript levels from 8 to 24 days post treatment. Downregulation of target genes ranged from 2- to 16-fold compared to the corresponding dsGFP controls, where highest silencing effects were generally noticed for ATP synthase beta. Sequencing of libraries obtained from total smallRNA (sRNA) showed the generation of dsRNA-derived sRNAs by RNAi pathway, with majority of reads mapping exclusively on the correspondent dsRNA. Also, we characterized components of a functional RNAi machinery in P. spumarius. Further research is needed to clarify such mechanism, screen effective target lethal genes to reduce vector population and improve delivery strategies.
Identifiants
pubmed: 39468170
doi: 10.1038/s41598-024-73889-5
pii: 10.1038/s41598-024-73889-5
doi:
Substances chimiques
RNA, Double-Stranded
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
25812Subventions
Organisme : Italian Ministry of Agriculture, MASAF
ID : SOS Project
Informations de copyright
© 2024. The Author(s).
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