Sequenciamento e análise comparativa do DNA de Cyphoderus similis FOLSOM e Cyphoderus innominatus MILLS

Abstract

The genus Cyphoderus Nicolet 1842 is the largest in the Paronellidae subfamily, with worldwide distribution. In Brazil there are nine species and among them there are Cyphoderus similis Folsom 1927 and Cyphoderus innominatus Mills 1938. Currently only four Collembola Class species have their complete genome published while only 14 species have their accessible mitochondrial genome. Studies involving DNA sequencing broaden knowledge of group biology at the molecular level, becoming a starting point for genetic, evolutionary, phylogenetic, and conservation studies. Therefore this dissertation has as main objective to sequence and compare the nuclear and mitochondrial DNA of C. similis and C. innominatus, and as specific objectives we have: the structural and functional description of the sequenced genomes; the comparison of sequenced genomes with other published genomes; perform a molecular phylogenetic test using the sequenced genes. The species C. similis was randomly collected from iron cave soils in Minas Gerais, while the species C. innominatus was collected from green areas of Paraíba State University, João Pessoa. Subsequently the specimens were kept in cultures, separated by species. At Vale Technological Institute - Sustainable Development (ITV-DS / PA), DNA extraction and sequencing steps were performed. The DNeasy Blood & Tissue extraction kit and Illumina NextSeq and MiniSeq sequencing technologies were used with paired-end and mate-pair techniques. The result is the sequencing of mitochondrial DNA with a total length of 14,988 bp with 36 genes for C. similis and 14,811 bp with 37 genes for C. innominatus, both presenting complete gene content and similar to other Collembola mitochondrial genomes. Phylogeny based on mitochondrial DNA sequences corroborates previous studies, but brings greater robustness to Collembola's current molecular phylogeny. Nuclear genomes contain a maximum total length of 358 Mb with N50 of 3.3 Kb for C. similis and 370 Mb with N50 of 3.2 Kb for C. innominatus. The gene annotation showed 278,865 putative genes found in the C. innominatus genome with a density of 3.8 genes for each exon found. Nuclear genomes are still fragmented and low in N50 when compared to other available genomes. Therefore, we conclude here the two new complete mitochondrial genomes, in addition to the first Nuclear Drafts Genomes for Cyphoderus similis and Cyphoderus innominatus, the first Collembola genomes for the Brazilian region.