| Congresso Brasileiro de Microbiologia 2023 | Resumo: 119-1 | ||||
Resumo:Entomopathogenic fungi are found worldwide and Metarhizium anisopliae is considered a model for studying interactions between fungi and insects. These fungi provide a promising means of control because they can infect and kill a range of arthropod hosts, including agricultural pests. The infection by entomopathogenic fungi begins when spores are deposited on the insect’s surface. The spores germinate and produce enzymes that degrade the insect’s cuticle, which is composed of proteins and chitin. In addition to these enzymes, secondary metabolites (SMs) also play a role in the infection process by damaging the host’s physiology or modulating its microbiota. In microorganisms, genes that code for proteins involved in the synthesis, maturation, and transport of secondary metabolites (SMs) are often arranged in biosynthetic gene clusters (BGCs), allowing the inference of co-expression. To evaluate BGC genes potentially associated with the virulence of M. anisopliae, we collected large-scale gene expression data (RNA-Seq) from the NCBI SRA repository and used it to construct normalized expression tables with the STAR aligner. We then used the WGCNA package to construct co-expression clusters and assess their correlation with different experimental conditions, including infection of tick and mimicked infection conditions (hyphae grown in grasshopper hemolymph and in wings. Four clusters - black, red, yellow, and greenyellow - had high correlation values of eigengenes with different infection conditions (r values of 0.783, 0.954, 0.587, and 0.706, respectively). To evaluate whether this expression was reflected in other insects, we performed RT-qPCR analyses of the backbone genes of six BGCs in conditions of infection of the insect Spodoptera frugiperda. Of these, two showed statistically significant increased expression levels compared to control conditions: MaPKS19 (MANI_012054). As a control, we also evaluated the expression of backbone genes present in one BGC known to be expressed during infection of the tick Ripichephalus microplus (MaPKS16 - MANI_029455 and MANI_029453). These results suggest that our protocol can be used to identify BGCs important for the virulence of Metarhizium species
Palavras-chave: entomopathogenic fungi, Metarhizium anisopliae, secondary metabolites, BGC Agência de fomento:Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). | |||||