TY - JOUR
T1 - Novel bacterial plasmid produces small interfering RNAs (siRNAs) that induce effective gene silencing in the Asian citrus psyllid Diaphorina citri
AU - Rueda-Silva, Juan Carlos
AU - González-Campos, Laura Isabel
AU - Durán-Armenta, Luis Fernando
AU - Karam-Coppola, Arantxa
AU - Antonio-Pérez, Aurora
AU - Ordoñez-Rodríguez, Jorge
AU - Saucedo-Tavitas, Nicole Estefanía
AU - Rico-Torres, Valeria
AU - Cruz-Cruz, Carlos
AU - García-Huante, Yolanda Guadalupe
AU - Amador-Molina, Juan Carlos
AU - Tapia-Ramírez, José Isabel
AU - Torres-Huerta, Ana Laura
N1 - Publisher Copyright:
© 2023 Pontificia Universidad Católica de Valparaíso
PY - 2023/7
Y1 - 2023/7
N2 - 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.
AB - 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.
KW - Asian citrus psyllid
KW - Candidatus Liberibacter asiaticus
KW - Citrus industry
KW - Diaphorina citri
KW - dsRNA-producing plasmid
KW - Gene silencing
KW - Huanglongbing disease
KW - Novel bacterial plasmid
KW - Pest control
KW - siRNA
KW - Small interfering RNAs
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UR - http://www.scopus.com/inward/citedby.url?scp=85162118885&partnerID=8YFLogxK
U2 - 10.1016/j.ejbt.2023.03.006
DO - 10.1016/j.ejbt.2023.03.006
M3 - Article
AN - SCOPUS:85162118885
SN - 0717-3458
VL - 64
SP - 59
EP - 68
JO - Electronic Journal of Biotechnology
JF - Electronic Journal of Biotechnology
ER -