Phylogenetic analysis of ITS sequences in Escovopsis isolates from Misiones, in the framework of a polyphasic identification

BARENGO M. P.; ALZAGA E. E.; BICH G. A.; AMERIO N. S.; ZAPATA P. D.; CASTRILLO M. L.

Congreso; 3rd WOMEN IN BIOINFORMATICS & DATA SCIENCE LA Fostering collaboration among women; 2022

WOMEN IN BIOINFORMATICS & DATA SCIENCE LA Fostering collaboration among women

In Misiones, one of the main pests that affects the primary forest sector are leaf cutterants. Fungi of Escovopsis genus (Ascomycota: Hypocreales), are considered possiblebiocontrol agents as specialist parasites of Leucoagaricus, the main food of these ants.The initial step in biocontrol strategies is the correct identification of fungal isolatesapplied as biocontrol agents. The ITS1-5.8S-ITS2 regions of ribosomal DNA areconsidered the primary universal barcode for fungi, which allow phylogeneticcomparison between species. Our group worked with 4 Escovopsis isolates, coded asHMP2, HMP8, HMP9 and HMP10; obtained from leaf cutter ant nests in Misiones. Inprevious macro-micromorphological analyses there were established that HMP2,HMP9, and HMP10 belonged to the Escovopsis primorosea species and HMP8 toEscovopsis catenulate species. The proposed objective was to carry out thephylogenetic analysis of the 4 isolates, within the framework of a polyphasicidentification. The ITS1-5.8S-ITS2 regions were amplified and sequenced, using theuniversal primers ITS1 and ITS4. The sequences obtained, together with sequencesretrieved from the NCBI-GenBank, were phylogenetically analyzed using the MEGA6software. The trees were made by the Neighbor-Joining (NJ) and Maximum Likelihood(ML) methods using the Bootstrap test with 1000 replications. The analysis of thesequences allowed the construction of two trees by the NJ and ML methods. IsolatesHMP2, HMP9, and HMP10 were clustered together, with a bootstrap of 82 for NJ and86 for ML; and HMP8 was placed in a separate clade, without clustering with referencesequences. This was because there are no ITS sequences published for the species E.catenulata and E. primorosea. Therefore, the phylogenetic relationships correspondedwith the morphological analyses, and these were the first sequences reported for thesespecies in the Genbank database, which will be relevant in further molecular orpolyphasic identification studies.