Desenvolvimento de marcadores microssatélites: painel de primers para Leiarius marmoratus através de sequenciamento de próxima geração.

Abstract

Leiarius marmoratus, popularly known as “Amazon jundiá”, has great aquaculture potential in genetic improvement programs to increase zootechnical performance related to hybrid vigor. Fish farming stations have commercially exploited the hybrid “pintado da-amazonia” obtained by crossing female Pseudoplatystoma punctifer x male Leiarius marmoratus. Therefore, genomic investigations are crucial for further exploration of L. marmoratus for the development of technological packages. Thus, the objective was to develop microsatellite markers and design primers for L. marmoratus as a tool for population studies and genetic improvement. Initially, a high molecular weight DNA sample was submitted to next generation sequencing by the Illumina platform, using the HiSeqTM system. The sequencing product was evaluated by the FastQC program, data filtering was performed for quality control using trimommetc. The de novo assembly of the genome was carried out in the SOAPdenovo2 and SPADES programs, the quality of the products generated was verified by the QUAST program. To identify the microsatellite markers, the MISA software and the EasySSR software were used; finally, BatchPrimer3 was used to design primers with the program's flow parameters. Therefore, an assembly with 225,027 contigs was obtained, a total of 221,273 SSRS motifs identified by MISA, with a higher frequency of pentanucleotides, differentiating from the EasySSR program which obtained 145,601 microsatellites, with a predominance of dinucleotides, also obtaining 590 successful primer pairs. successful. Proving the excellent usefulness of molecular methodologies for obtaining biotechnological tools such as SSRs, contributing to population genetic studies, controlling genetic variability, mapping, genomic and comparative analysis.