The mycoparasitic fungus Clonostachys pityrodes: phylogenetic analysis as a tool for molecular identification

Abstract

Biological control is a promising and sustainable strategy to reduce damage caused by agricultural pests and the use of chemical fungicides. Fungal strains of the genus Clonostachys are studied as biocontrol agent of fungi and nematodes. However, the presence of this fungus in the soils of Misiones remains unexplored. Traditional fungal identification is generally carried out by morphological characterization in Petri dishes, and by observing their reproductive structures under the microscope. In general, with this methodology it is possible to identify to the genus level, however determining up to the species level is usually very complicated in some genera and many times ambiguities are achieved. In this context, molecular data emerges as an important tool to complement morphological information and thus achieve a correct fungal identification. The objective of this work was to molecularly identify with ITS markers a strain of the mycoparasitic fungus Clonostachys HEP30. The nucleic acids were isolated for molecular corroboration. From the extracted genetic material, the ITS1-5,8S-ITS2 region was amplified and sequenced. Once the region of interest was obtained, the information obtained was compared with that existing in the databases, using the Blast (Basic Local Alignment Search Tool) of the NCBI (National Center for Biotechnology Information) and the fungal barcoding database and then phylogenetic analysis was done. The molecular identification and phylogenetic analysis allowed us to classify the fungal isolate Clonotachys HEP 30 with high percentage of identity as a member of Clonostachys pityrodes species.