Transcriptome analysis and exploring RNAi and CRISPR in the Neotropical stink bug, Euschistus heros

• Autor(a) principal: Cagliari, Deise
• Data de Publicação: 2021
• Tipo de documento: Tese
• Idioma: eng
• Título da fonte: Repositório Institucional da UFPel - Guaiaca
• Texto Completo: http://guaiaca.ufpel.edu.br/handle/prefix/8383
• topic: CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE; Gene silencing; Gene knockdown; Gene knockout; Pest management; Functional genomics; Silenciamento gênico; Manejo de insetos-praga; Genética funcional

One of the main factors limiting agricultural production is the attack of insects, pathogens and weeds, which can cause significant losses in the culture. Euschistus heros (Heteroptera: Pentatomidae) is one of the most important stink bug species found in soybean, with occurrence distributed throughout the Brazilian production area. In an attempt to reduce the damage caused by this and other insects, producers depend almost exclusively on the use of insecticides. However, the indiscriminate use of these products has been causing serious management problems, such as the selection of resistant populations leading to the inefficiency of products available on the market. Therefore, the development of alternative control tools, such as gene silencing based on RNA interference (RNAi) and CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is necessary. RNAi is a highly conserved mechanism in eukaryotic organisms and by which the messenger RNA molecule (mRNA) is cleaved by the gene-silencing machinery, leading to inactivation of gene expression (Knockdown). CRISPR, on the other hand, is a gene editing tool with which it is possible to carry out modification at the DNA level, introducing or eliminating specific DNA sequences from the genome (knockout). During the last years, the use of RNAi and CRISPR has attracted the interest of researchers, mainly in the study of the functions of genes during insect’s development and reproduction, which will allow the future development of alternative control strategies. Thus, considering the importance of RNAi and the CRISPR / Cas9 system in the study of functional genomics and in the development of new tools for the management of insect pests, the objectives of this study include: i) to identify the main components of the RNAi machinery present in E. heros, as well as validating the functionality of this tool in E. heros; ii) a review showing the potential use of the RNAi tool in a non-transformative approach; iii) the use of parental RNAi as a tool in the study of gene functions in E. heros; iv) the combination of RNAi and CRISPR/Cas9 as functional genomic tools in the Neotropical bug, E. heros. This information provides novel and important dataset on RNAi machinery and its efficiency, the future perspective of non-transformative RNAi approaches and the use of RNAi and CRISPR as tools in functional genetic studies, underpinning future strategies to improve RNAi and CRISPR in E. heros and other piercing-sucking insects species important in agriculture.