RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli
BACKGROUND: The Mediterranean fruit fly (medfly), Ceratitis capitata Wiedemann, is a major pest affecting fruit and vegetable production worldwide, whose control is mainly based on insecticides. Double-stranded RNA (dsRNA) able to down-regulate endogenous genes, thus affecting essential vital functi...
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| Format: | Artículo |
| Language: | Inglés |
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Wiley Online Library
2023
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| Online Access: | https://hdl.handle.net/20.500.11939/8735 https://onlinelibrary.wiley.com/doi/full/10.1002/ps.7839 |
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| author | Ortolá, Beltrán Urbaneja, Alberto Eiras, Marcelo Pérez-Hedo, Mertixell Daros, José A. |
| author_browse | Daros, José A. Eiras, Marcelo Ortolá, Beltrán Pérez-Hedo, Mertixell Urbaneja, Alberto |
| author_facet | Ortolá, Beltrán Urbaneja, Alberto Eiras, Marcelo Pérez-Hedo, Mertixell Daros, José A. |
| author_sort | Ortolá, Beltrán |
| collection | ReDivia |
| description | BACKGROUND: The Mediterranean fruit fly (medfly), Ceratitis capitata Wiedemann, is a major pest affecting fruit and vegetable production worldwide, whose control is mainly based on insecticides. Double-stranded RNA (dsRNA) able to down-regulate endogenous genes, thus affecting essential vital functions via RNA interference (RNAi) in pests and pathogens, is envisioned as a more specific and environmentally-friendly alternative to traditional insecticides. However, this strategy has not been explored in medfly yet. RESULTS: Here, we screened seven candidate target genes by injecting in adult medflies gene-specific dsRNA hairpins transcribed in vitro. Several genes were significantly down-regulated, resulting in increased insect mortality compared to flies treated with a control dsRNA targeting the green fluorescent protein (GFP) complementary DNA (cDNA). Three of the dsRNAs, homologous to the beta subunit of adenosine triphosphate (ATP) synthase (ATPsynbeta), a vacuolar ATPase (V-ATPase), and the ribosomal protein S13 (RPS13), were able to halve the probability of survival in only 48 h after injection. We then produced new versions of these three dsRNAs and that of the GFP control as circular molecules in Escherichia coli using a two-self-splicing-intron-based expression system and tested them as orally-delivered insecticidal compounds against medfly adults. We observed a significant down-regulation of V-ATPase and RPS13 messenger RNAs (mRNAs) (approximately 30% and 90%, respectively) compared with the control medflies after 3 days of treatment. No significant mortality was recorded in medflies, but egg laying and hatching reduction was achieved by silencing V-ATPase and RPS13. CONCLUSION: In sum, we report the potential of dsRNA molecules as oral insecticide in medfly. |
| format | Artículo |
| id | ReDivia8735 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | Wiley Online Library |
| publisherStr | Wiley Online Library |
| record_format | dspace |
| spelling | ReDivia87352025-04-25T14:49:24Z RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli Ortolá, Beltrán Urbaneja, Alberto Eiras, Marcelo Pérez-Hedo, Mertixell Daros, José A. Medfly RNA interference Doubled-stranded RNA Self-splicing intron Vacuolar ATPase Ribosomal protein S13 Alternative to insecticides H10 Pests of plants U30 Research methods Ceratitis capitata Escherichia coli Pest control BACKGROUND: The Mediterranean fruit fly (medfly), Ceratitis capitata Wiedemann, is a major pest affecting fruit and vegetable production worldwide, whose control is mainly based on insecticides. Double-stranded RNA (dsRNA) able to down-regulate endogenous genes, thus affecting essential vital functions via RNA interference (RNAi) in pests and pathogens, is envisioned as a more specific and environmentally-friendly alternative to traditional insecticides. However, this strategy has not been explored in medfly yet. RESULTS: Here, we screened seven candidate target genes by injecting in adult medflies gene-specific dsRNA hairpins transcribed in vitro. Several genes were significantly down-regulated, resulting in increased insect mortality compared to flies treated with a control dsRNA targeting the green fluorescent protein (GFP) complementary DNA (cDNA). Three of the dsRNAs, homologous to the beta subunit of adenosine triphosphate (ATP) synthase (ATPsynbeta), a vacuolar ATPase (V-ATPase), and the ribosomal protein S13 (RPS13), were able to halve the probability of survival in only 48 h after injection. We then produced new versions of these three dsRNAs and that of the GFP control as circular molecules in Escherichia coli using a two-self-splicing-intron-based expression system and tested them as orally-delivered insecticidal compounds against medfly adults. We observed a significant down-regulation of V-ATPase and RPS13 messenger RNAs (mRNAs) (approximately 30% and 90%, respectively) compared with the control medflies after 3 days of treatment. No significant mortality was recorded in medflies, but egg laying and hatching reduction was achieved by silencing V-ATPase and RPS13. CONCLUSION: In sum, we report the potential of dsRNA molecules as oral insecticide in medfly. 2023-11-15T07:36:22Z 2023-11-15T07:36:22Z 2023 article publishedVersion Ortolá, B., Urbaneja, A., Eiras, M., Pérez‐Hedo, M., & Daròs, J. A. (2024). RNAi‐mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally‐supplied double‐stranded RNAs produced in Escherichia coli. Pest Management Science, 80(3), 1087-1098. 1526-4998 (eISSN) 1526-498X (Print ISSN) https://hdl.handle.net/20.500.11939/8735 10.1002/ps.7839 https://onlinelibrary.wiley.com/doi/full/10.1002/ps.7839 en This work was supported by grants PID2020-114691RB-I00 and PID2020-113234RR-I00 from the Spanish Ministerio de Ciencia e Innovación through the Agencia Estatal de Investigación, and by grants IVIA-52202B and IVIA-51916 from Instituto Valenciano de Investigaciones Agrarias (both projects are susceptible of being co-financed by the European Union through the European Regional Development Fund Operational Program). BO was the recipient of a predoctoral contract from Universitat Politècnica de València (PAID-01-17). info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-113234RR-I00/ES/EXPOSICION A VOLATILES INDUCIDOS POR PLANTAS: UNA VIA SOSTENIBLE DE MANEJO DE PLAGAS Y ENFERMEDADES A TRAVES DE LA INDUCCION DEFENSIVA DE LAS PLANTAS info:eu-repo/grantAgreement/AEI/Programa Estatal de I+D+i Orientada a los Retos de la Sociedad/PID2020-114691RB-I00/ES/BIOTECNOLOGIA DE VIRUS DE PLANTAS: VECTORES VIRALES Y ESTRATEGIAS DE RESISTENCIA info:eu-repo/grantAgreement/ERDF/PCV 2021-2027/52202/ES/Control biológico de plagas y enfermedades como pieza clave en el impulso de la Sostenibilidad de los cultivos agrarios de la Comunidad Valenciana/SOSTENIBLE Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ openAccess Wiley Online Library electronico |
| spellingShingle | Medfly RNA interference Doubled-stranded RNA Self-splicing intron Vacuolar ATPase Ribosomal protein S13 Alternative to insecticides H10 Pests of plants U30 Research methods Ceratitis capitata Escherichia coli Pest control Ortolá, Beltrán Urbaneja, Alberto Eiras, Marcelo Pérez-Hedo, Mertixell Daros, José A. RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli |
| title | RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli |
| title_full | RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli |
| title_fullStr | RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli |
| title_full_unstemmed | RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli |
| title_short | RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli |
| title_sort | rnai mediated silencing of mediterranean fruit fly ceratitis capitata endogenous genes using orally supplied double stranded rnas produced in escherichia coli |
| topic | Medfly RNA interference Doubled-stranded RNA Self-splicing intron Vacuolar ATPase Ribosomal protein S13 Alternative to insecticides H10 Pests of plants U30 Research methods Ceratitis capitata Escherichia coli Pest control |
| url | https://hdl.handle.net/20.500.11939/8735 https://onlinelibrary.wiley.com/doi/full/10.1002/ps.7839 |
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