Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri
Background: The Asian citrus psyllid, Diaphorina citri, is a vector of the plant pathogen Candidatus Liberibacter asiaticus (CLas), the causal agent of Huanglongbing disease. HLB represents the main threat to the citrus industry around the world due to its fast spreading, high infectivity, and incur...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-07-01
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| Series: | Electronic Journal of Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345823000179 |
| _version_ | 1797786564945772544 |
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| author | Juan Carlos Rueda-Silva Laura Isabel González-Campos Luis Fernando Durán-Armenta Arantxa Karam-Coppola Aurora Antonio-Pérez Jorge Ordoñez-Rodríguez Nicole Estefanía Saucedo-Tavitas Valeria Rico-Torres Carlos Cruz-Cruz Yolanda Guadalupe García-Huante Juan Carlos Amador-Molina José Isabel Tapia-Ramírez Ana Laura Torres-Huerta |
| author_facet | Juan Carlos Rueda-Silva Laura Isabel González-Campos Luis Fernando Durán-Armenta Arantxa Karam-Coppola Aurora Antonio-Pérez Jorge Ordoñez-Rodríguez Nicole Estefanía Saucedo-Tavitas Valeria Rico-Torres Carlos Cruz-Cruz Yolanda Guadalupe García-Huante Juan Carlos Amador-Molina José Isabel Tapia-Ramírez Ana Laura Torres-Huerta |
| author_sort | Juan Carlos Rueda-Silva |
| collection | DOAJ |
| description | Background: The Asian citrus psyllid, Diaphorina citri, is a vector of the plant pathogen Candidatus Liberibacter asiaticus (CLas), the causal agent of Huanglongbing disease. HLB represents the main threat to the citrus industry around the world due to its fast spreading, high infectivity, and incurability. An alternative for the control of this plague in Rutaceae is the implementation of RNA interference (RNAi). Here, we propose a novel method to produce small interfering RNA (siRNA) in Escherichia coli. The method includes a reporter system to visually demonstrate dsRNA expression. We produced and tested siRNAs against three D. citri genes: Abnormal Wing Disk (AWD), Superoxide dismutase 1 (SOD), and Wingless (WNT). These genes play key roles in psyllid development and maturity, and thus, represent promising targets for potential vector control. Results: The blue coloration in bacterial cultures was easily observable, and it corroborated the generation of the dsRNA that is in the same transcript. We evaluated the dosage efficiency of siRNA using the AWD siRNA. The RNAi treatment was evaluated, and all the siRNAs tested were able to induce silencing (-3.05 for AWD, -2.60 for SOD and -2.57 for WNT). Conclusions: The novel bacterial plasmid effectively produces siRNAs, and the blue color reporter is visually facilitated to check the expression of each dsRNA. Treating D. citri with the produced siRNAs resulted in a decrease in gene expression. In addition, psyllid mortality was observed, being the highest when treated with WNT-siRNA. Our results suggest the potential of siRNA treatment as a method for controlling Huanglongbing.How to cite: Rueda-Silva JC, González-Campos LI, Durán-Armenta LF, et al. Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri. Electron J Biotechnol 2023;64. https://doi.org/10.1016/j.ejbt.2023.03.006. |
| first_indexed | 2024-03-13T01:09:43Z |
| format | Article |
| id | doaj.art-5e5842c5feb7493cb8d18075931b584b |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-03-13T01:09:43Z |
| publishDate | 2023-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-5e5842c5feb7493cb8d18075931b584b2023-07-06T04:17:14ZengElsevierElectronic Journal of Biotechnology0717-34582023-07-01645968Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citriJuan Carlos Rueda-Silva0Laura Isabel González-Campos1Luis Fernando Durán-Armenta2Arantxa Karam-Coppola3Aurora Antonio-Pérez4Jorge Ordoñez-Rodríguez5Nicole Estefanía Saucedo-Tavitas6Valeria Rico-Torres7Carlos Cruz-Cruz8Yolanda Guadalupe García-Huante9Juan Carlos Amador-Molina10José Isabel Tapia-Ramírez11Ana Laura Torres-Huerta12Departamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Genética y Biologı́a Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Ciudad de México C.P. 07360, MéxicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, MexicoDepartamento de Genética y Biologı́a Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Ciudad de México C.P. 07360, México; Corresponding authors.Departamento de Bioingeniería, Tecnológico de Monterrey, Campus Estado de México, Av. Lago de Guadalupe KM 3.5, Margarita Maza de Juárez, Cd. López Mateos, Atizapán de Zaragoza 52926, Mexico; Corresponding authors.Background: The Asian citrus psyllid, Diaphorina citri, is a vector of the plant pathogen Candidatus Liberibacter asiaticus (CLas), the causal agent of Huanglongbing disease. HLB represents the main threat to the citrus industry around the world due to its fast spreading, high infectivity, and incurability. An alternative for the control of this plague in Rutaceae is the implementation of RNA interference (RNAi). Here, we propose a novel method to produce small interfering RNA (siRNA) in Escherichia coli. The method includes a reporter system to visually demonstrate dsRNA expression. We produced and tested siRNAs against three D. citri genes: Abnormal Wing Disk (AWD), Superoxide dismutase 1 (SOD), and Wingless (WNT). These genes play key roles in psyllid development and maturity, and thus, represent promising targets for potential vector control. Results: The blue coloration in bacterial cultures was easily observable, and it corroborated the generation of the dsRNA that is in the same transcript. We evaluated the dosage efficiency of siRNA using the AWD siRNA. The RNAi treatment was evaluated, and all the siRNAs tested were able to induce silencing (-3.05 for AWD, -2.60 for SOD and -2.57 for WNT). Conclusions: The novel bacterial plasmid effectively produces siRNAs, and the blue color reporter is visually facilitated to check the expression of each dsRNA. Treating D. citri with the produced siRNAs resulted in a decrease in gene expression. In addition, psyllid mortality was observed, being the highest when treated with WNT-siRNA. Our results suggest the potential of siRNA treatment as a method for controlling Huanglongbing.How to cite: Rueda-Silva JC, González-Campos LI, Durán-Armenta LF, et al. Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri. Electron J Biotechnol 2023;64. https://doi.org/10.1016/j.ejbt.2023.03.006.http://www.sciencedirect.com/science/article/pii/S0717345823000179Asian citrus psyllidCandidatus Liberibacter asiaticusCitrus industryDiaphorina citridsRNA-producing plasmidGene silencing |
| spellingShingle | Juan Carlos Rueda-Silva Laura Isabel González-Campos Luis Fernando Durán-Armenta Arantxa Karam-Coppola Aurora Antonio-Pérez Jorge Ordoñez-Rodríguez Nicole Estefanía Saucedo-Tavitas Valeria Rico-Torres Carlos Cruz-Cruz Yolanda Guadalupe García-Huante Juan Carlos Amador-Molina José Isabel Tapia-Ramírez Ana Laura Torres-Huerta Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri Electronic Journal of Biotechnology Asian citrus psyllid Candidatus Liberibacter asiaticus Citrus industry Diaphorina citri dsRNA-producing plasmid Gene silencing |
| title | Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri |
| title_full | Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri |
| title_fullStr | Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri |
| title_full_unstemmed | Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri |
| title_short | Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri |
| title_sort | novel bacterial plasmid produces small interfering rnas sirnas that induce effective gene silencing in the asian citrus psyllid diaphorina citri |
| topic | Asian citrus psyllid Candidatus Liberibacter asiaticus Citrus industry Diaphorina citri dsRNA-producing plasmid Gene silencing |
| url | http://www.sciencedirect.com/science/article/pii/S0717345823000179 |
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