Authors

Matteo Ripamonti,1,2 Luca Cerone,1,3 Simona Abbà,1 Marika Rossi,1 Sara Ottati,1,3 Sabrina Palmano,1 Cristina Marzachì,1 and Luciana Galetto1,*

Affiliations

"1Istituto per la Protezione Sostenibile Delle Piante, Consiglio Nazionale Delle Ricerche, IPSP-CNR, Strada delle Cacce 73, 10135 Torino, Italy; [email protected] (M.R.); [email protected]_enorec (L.C.); [email protected] (S.A.); [email protected] (M.R.); [email protected] (S.O.); [email protected] (S.P.); [email protected] (C.M.)
2Environmental Research and Innovation Department (ERIN), Luxembourg Institute of Science and Technology (LIST), 41 Rue du Brill, 4422 Luxembourg, Luxembourg
3Dipartimento di Scienze Agrarie, Forestali ed Alimentari DISAFA, Università degli Studi di Torino, Largo Paolo Braccini 2, Grugliasco, 10095 Torino, Italy
*Correspondence: [email protected]"

Topic

Cell Biology

Abstract

"Scaphoideus titanus (Hemiptera: Cicadellidae) is the natural vector of Flavescence dorée phytoplasma, a quarantine pest of grapevine with severe impact on European viticulture. RNA interference (RNAi) machinery components are present in S. titanus transcriptome and injection of ATP synthase β dsRNAs into adults caused gene silencing, starting three days post injection (dpi) up to 20 dpi, leading to decrease cognate protein. Silencing of this gene in the closely related leafhopper Euscelidius variegatus previously showed female sterility and lack of mature eggs in ovaries. Here, alteration of developing egg morphology in S. titanus ovaries as well as overexpression of hexamerin transcript (amino acid storage protein) and cathepsin L protein (lysosome proteinase) were observed in dsATP-injected females. To evaluate RNAi-specificity, E. variegatus was used as dsRNA-receiving model-species. Different doses of two sets of dsRNA-constructs targeting distinct portions of ATP synthase β gene of both species induced silencing, lack of egg development, and female sterility in E. variegatus, indicating that off-target effects must be evaluated case by case. The effectiveness of RNAi in S. titanus provides a powerful tool for functional genomics of this non-model species and paves the way toward RNAi-based strategies to limit vector population, despite several technical and regulatory constraints that still need to be overcome to allow open field application.

Keywords: RNA interference, hexamerin, cathepsin L, RNAi specificity, Euscelidius variegatus, female sterility

DOI: https://dx.doi.org/ 10.3390/ijms23020765

Extract

… observed under a DM750 (Leica Wetzlar, Germany) microscope equipped with a CoolLED pE300 white Illumination System.

Product Associated Features

The pE-300white is a popular illuminator for everyday fluorescent screening and analysis with simple operatation and individual irradiance control of each LED channel.

Product Type

pE-300white

Journal

International Journal of Molecular Sciences

Year of Publication

2022

Country of Publication

Switzerland