Identificação molecular de helicoverpa armigera (Lepidoptera: Noctuidae)

• Autor(a) principal: Oliveira, Thayssa Monize Rosa de
• Data de Publicação: 2021
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Repositório Institucional da UFG
• Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/12678
• topic: Lagarta do velho mundo; PCR; Primers específicos; Old world bollworm; Specie-specific primers; CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE

Previously found only on Old World continents, the caterpillar Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is capable of great economic damage, coming to be considered one of the most destructive agricultural pests. Its identification through morphological analysis is only possible with complex dissection of the adult male genitalia. The morphology of all phases of the life cycle, egg, larva, pupa and adult, is very similar with the morphology of other species of the genus, making the identification difficult. As it is a highly invasive pest and correct identification is the first step to be taken to decide the best management strategy, we realize the need for precise, easy and fast tools for the identification of this species. Thus, the objective of the work was to develop specific primers and to optimize different protocols for use in real-time PCR and digital PCR methodologies in order to identify H. armigera, even if this sample is of poor quality, only a fragment and/or is mixed with other species. For that, design, selection and optimization of primer and probe concentrations, primer annealing temperature gradients, tests and adaptations with different DNA extraction methods were carried out, with or without post-DNA extraction purification, tests with different ratios of H. zea and H. armigera mixed in the same reaction, determination of the maximum sensitivity in both methodologies and interpretation of the obtained data. We present the protocols developed to successfully identify the specie H. armigera, both with real-time PCR and digital PCR methodologies.