Konfirmasi Spesies Trichoderma Spp. dengan Metode Molekuler

Abstract

Trichoderma spp. is a soil fungus that acts as a biological agent due to its antagonistic properties against pathogens. Conventional morphological identification of Trichoderma fungi is often insufficient to accurately distinguish species, so a molecular approach is needed as a complementary method. This study aims to confirm Trichoderma spp. species through molecular methods. All Trichoderma isolates used were obtained from the AM2B Laboratory and PT.BGA collections. Macroscopic observation shows that the colony is spherical in shape with a dominant color ranging from whitish green to dark green and white edges, as well as a fine hairy mycelium texture. Microscopically, Trichoderma conidiophores are known to be branched, phialides are cylindrical to curved, and conidia are round to oval, which are characteristic features of the Trichoderma genus. The molecular method was carried out with the stages of DNA extraction, PCR amplification using ITS5/ITS4 primers, agarose gel electrophoresis, and DNA fragment size analysis. ITS amplification successfully produced DNA fragments measuring 670 bp in lane 1, 613 bp in lane 2, and approximately 600 bp in lanes 3 and 4. The size of the fragments obtained was within the range of ITS amplicons characteristic of the Trichoderma genus. However, based on the results of amplification with specific primers for the tef1-α and rpb2 genes, the isolates in these samples did not belong to the species T. virens, T. asperellum, T. longibrachiatum, or T. harzianum.