PHYLOGENETIC ANALYSIS OF ITS SEQUENCES FROM ISOLATES OF ESCOVOPSIS, NATIVES OF THE PROVINCE OF MISIONES

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

Congreso; 1st Latin American Congress of Women in Bioinformatics & Data Science LA.; 2020

Latin American Congress of Women in Bioinformatics & Data Science LA.

In Misiones, one of the main pests that affect forestry are the leaf cutter ants. Fungi of the Escovopsis genus are considered possible biocontrol agents since they are capable of parasitizing Leucoagaricus, the main food of these ants.The initial step in any biocontrol strategy is the correct identification of fungal isolates. The ITS1-5,8S-ITS2 regions of the ribosomal DNA are considered the primary universal barcode for fungi, allowing the phylogenetic comparison between species.Our working group isolated three Escovopsis isolates, coded as HMP1, HMP5 and HMP6 obtained from nests of leaf cutter ants from Misiones. In previous macro-micromorphological analyzes, it was possible to establish that HMP1 belonged to E. microspora species and, HMP5 and HMP6 to E. weberi species. The aim was to perform the molecular phylogenetic analysis of the three Escovopsis isolates in order to complete the polyphasic identification. The ITS1-5,8S-ITS2 regions were amplified and sequenced, using the universal primers ITS1 and ITS4. The sequences obtained, together with sequences retrieved from NCBI-GenBank, were analyzed phylogenetically using MEGA6 software. The trees were constructed by the Neighbor-Joining (NJ) and Maximum Likelihood (ML) methods using the Bootstrap test with 1000 replicates.The sequence analysis allowed the construction of two phylogenetic trees, in which the three analyzed sequences were grouped in the same monophyletic clade with E. microspora and E. weberi, with a Bootstrap of 85, for NJ and ML, showing a close relationship between the ITS sequences of both species. The trees made allowed corroborating the morphological identification, without being able to distinguish between the two species. Therefore, this work added molecular information about the polyphasic identificaction of these fungal isolates, but further studies with other molecular markers are need to separate the clades.