Abstract
AbstractOver the past few years, the use of RNA interference (RNAi) for insect pest management has attracted considerable interest in academia and industry as a pest-specific and environment-friendly strategy for pest control. For the success of this technique, the presence of core RNAi genes and a functional silencing machinery is essential. Therefore, the aim of this study was to test whether the Neotropical brown stinkbug Euschistus heros has the main RNAi core genes and whether the supply of dsRNA could generate an efficient gene silencing response. To do this, total mRNA of all developmental stages was sequenced on an Illumina platform, followed by a de novo assembly, gene annotation and RNAi-related gene identification. Once RNAi-related genes were identified, nuclease activities in hemolymph were investigated through an ex vivo assay. To test the functionality of the siRNA machinery, E. heros adults were microinjected with ~28 ng per mg of insect of a dsRNA targeting the V-ATPase-A gene. Mortality, relative transcript levels of V-ATPase-A, and the expression of the genes involved in the siRNA machinery, Dicer-2 (DCR-2) and Argonaute 2 (AGO-2), were analyzed. Transcriptome sequencing generated more than 126 million sequenced reads, and these were annotated in approximately 80,000 contigs. The search of RNAi-related genes resulted in 47 genes involved in the three major RNAi pathways, with the absence of sid-like homologous. Although ex vivo incubation of dsRNA in E. heros hemolymph showed rapid degradation, there was 35% mortality at 4 days after treatment and a significant reduction in V-ATPase-A gene expression. These results indicated that although sid-like genes are lacking, the dsRNA uptake mechanism was very efficient. Also, 2-fold and 4-fold overexpression of DCR-2 and AGO-2, respectively, after dsRNA supply indicated the activation of the siRNA machinery. Consequently, E. heros has proven to be sensitive to RNAi upon injection of dsRNA into its hemocoel. We believe that this finding together with a publically available transcriptome and the validation of a responsive RNAi machinery provide a starting point for future field applications against one of the most important soybean pests in South America.
Funder
Ministry of Science, Technology and Innovation | Conselho Nacional de Desenvolvimento Científico e Tecnológico
European Cooperation in Science and Technology
Publisher
Springer Science and Business Media LLC
Reference115 articles.
1. Medeiros, L. & Amegier, G. A. Ocorrência e Desempenho de Euschistus heros (F.) (Heteroptera: Pentatomidae) em Plantas Hospedeiras Alternativas no Rio Grande do Sul. Neotrop. Entomol. 38(4), 459–463 (2009).
2. Panizzi, A. R. S. B. Growing Problems with (Hemiptera: Heteroptera: Pentatomidae): Species Invasive to the U.S. and Potential. Am. Entomol. 61, 223–233 (2015).
3. Panizzi, A. R., Bueno, A. F. & Silva, F. A. C. Insetos que atacam vagens e grãos. Hoffmann-Campo, C. B., Corrêa-Ferreira, B. S. & Moscardi, F. (ed.) Soja: manejo integrado de insetos e outros artrópodes-praga. 5, 335–420. (Embrapa, 2012).
4. Panizzi, A. R. et al. Insetos da Soja no Brasil. Bol. Técnico n° 1 https://ainfo.cnptia.embrapa.br/digital/bitstream/item/77369/1/CNPSO-BOL.-TEC.-1-77.pdf (1977).
5. Sosa-Gómez, D. R. et al. Insecticide susceptibility of Euschistus heros (Heteroptera: Pentatomidae) in Brazil. J. Econ. Entomol. 102, 1209–1216 (2009).
Cited by
20 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献