Evaluación de marcadores moleculares para la discriminación de especies de Trichoderma spp. obtenidas de ecosistemas forestales.

Abstract

This study aimed to evaluate the different molecular markers for the discrimination of Trichoderma species obtained from forest ecosystems. 43 strains had to be reactivated, then the mycelium was extracted. The Wizard Genomic DNA Purification (Promega Corporation) kit protocol was followed for the DNA extraction process. Once the DNA was obtained, the PCR processes were performed based on the protocol of Zhou et. It means that for ITrichF/ITrichR (ITS): 96°C for 1:30 minutes, 35 cycles, 96°C for 35 seconds, 53°C for 55 seconds, 72°C for 1:30 minutes, 72°C for 10 minutes. The primers used for the ITS region in the genus Trichoderma were ITricF/ITricR with an approximate size of the PCR products from 103 to 113 bp, for the species the TEF1 gene was used, the primers were EHarF2/EHarR2 for T. harzianum and the primers used for Trichoderma longibrachiatum were T1F/T1R with an approximate size of the PCR products of 253 bp and 452 bp respectively, then the electrophoresis was performed. 7 isolates were chosen to verify whether they amplified according to the species studied and were sent to MACROGEN KOREA, the results were linked to the database of the National Center for Biotechnology Information using the BLAST method. 43 strains were studied, 31 of these ones within the genus Trichoderma. 17 and 12 strains were identified using different molecular markers for T. harzianum and T. longibrachiatum respectively. 100% coverage and identity were obtained of the 7 identified samples. It was concluded that the primers used effectively work for the discrimination of species in Trichoderma. It is recommended to replicate this study using the protocols that worked best in a larger number of isolates.