Analysis of the Gene Expression and RNAi-Mediated Knockdown of Chitin Synthase from Leaf-Cutting Ant Atta sexdens

Autor(es)

Ariele C. Moreira ; Renato L. Carneiro ^[2] ; Mariana F. Fracola ^[3] ; Kelli C. Micocci ^[4] ; Odair C. Bueno ^[5] ; Dulce H. F. Souza ^[6]

• Tipo de documento: Artigo Científico
• Fonte: Journal of the Brazilian Chemical Society; v. 31, n. 10, p. 1979-1990, 2020-10-09.

Resumo

Chitin synthase (CHS) is the enzyme specifically associated with chitin synthesis, an important component of diverse organisms including insects. Two alternative spliced transcripts of the CHS gene (AsCHS-A1 and AsCHS-A2) were identified by quantitative reverse-transcription polymerase chain reaction (RT-qPCR) during the development of the leaf-cutting ant Atta sexdens. Expression profiles of AsCHS-A transcripts increased from larva to pupae and decay in workers. Phylogenetic analysis showed both transcripts are classified within class A insect CHSs. AsCHS-A1 showed the highest expression level in larvae and pupae, while AsCHS-A2 is the main CHS transcript in workers. Our results suggest that these variants should be under regulation of different promoters. AsCHS-A1 has topology expected for insect CHSs, while the predicted AsCHS-A2 topology, with a missing A domain, is similar to some fungal CHSs. CHS-B (class B) was not identified in A. sexdens transcriptome. Ribonucleic acid interference (RNAi)-mediated gene silencing in pupae revealed that low reduction in CHS transcript levels (18%) was enough to cause morphological changes in the pupa exoskeleton impairing the process of cuticle sclerotization. To our knowledge, this work was the first to use and to show the feasibility of using RNA interference techniques on leaf-cutting ants. (AU)

• Processo FAPESP: 12/25299-6 - Estudos integrados para o controle de formigas cortadeiras
• Beneficiário: João Batista Fernandes
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/06198-8 - Estudo funcional de acetilcolinesterase (AChE) e outras enzimas de Atta Sexdens rubropilosa e seu fungo simbionte Leucoagaricus gongylophorus, imobilizações das mesmas em eletrodo e busca de inibidores específicos
• Beneficiário: Kelli Cristina Micocci
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/12169-2 - Estudos de vias metabólicas importantes no controle de formigas cortadeiras
• Beneficiário: Dulce Helena Ferreira de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Regular