Diversidade, relações filogenéticas e perfil de expressão gênica de transportadores MFS em Trichoderma spp

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2021-04-21

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Universidade Federal de Goiás

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The Trichoderma genus comprises filamentous fungi with mycoparasitism ability and high tolerance to abiotic stresses, such as the presence of metals. In both conditions, cellular transporters are vital to nutrients absorption and metallic ion extrusion. Among those, the Major Facilitator Superfamily (MFS) correspond to the largest superfamily of secondary transporters found in every organism, however a deep understanding of those is needed in Trichoderma spp. The present work aimed to identify the MFS transporters from 32 species of this genus, as well as to study their phylogenetic relationship and gene expression profiles. Global MFS identification were performed in silico using the Trichoderma spp. reference proteomes available on the Mycocosm database from Joint Genome Institute (JGI), by searching for annotated aminoacid sequences using the BlastP tool. The sequences were categorized in their respective families, according to reference sequences from the Transporter Classification Database (TCDB), and maximum likelihood phylogenetic trees were estimated for each of the families, using the IQTREE software. Lastly, MFS genes expression profiles were investigated during mycoparasitism and presence of metals, using RNA-Seq public data. 8,467 putative MFS were identified in Trichoderma spp., with a mean of 264 ± 51 sequences, and this number was higher than those found in other organisms. Mycoparasitic species presented a higher number of MFS. 55.5% of the sequences showed 12 transmembrane helices and 85.5% showed molecular weights between 50 and 70 kDa, with a mean of 58 kDa. In total, 24 MFS families were identified in the analyzed species, and the families regarding to nutrition, maintenance of homeostasis and cell defense showed a higher number of members. We did not observed enrichment of families in any species analyzed. In the phylogenetic inference the sequences grouped according to their functions and not to their lineage, moreover, SHS and GPH families showed subdivisions. In T. harzianum, besides a higher number of differently expressed genes during mycoparasitism, a higher number of upregulated genes was also observed in this condition compared to those in the presence of metals. In the presence of cadmium, we observed a higher downregulation of MFS genes than in the presence of aluminum. We also observed that ACS and DHA2 families were mostly differentially expressed during mycoparasitism 24 h, whilst FHS and PHS were mostly differentially expressed during the presence of aluminum 1.5 and 3.0 mg/mL, and MCT during the presence of cadmium 2.0 mg/mL. This work shed light on the description of MFS transporters and their functions during some of the biotic and abiotic stresses faced by Trichoderma spp. in the course of its development on environment.

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BARBOSA FILHO, J. R. Diversidade, relações filogenéticas e perfil de expressão gênica de transportadores MFS em Trichoderma spp. 2021. 96 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Goiás, Goiânia, 2021.