The current methodologies available for Trichoderma sp. multiplication under controlled conditions require long periods to produce the fungal inoculum. This study presents a simple and efficient methodology for multiplication of pure Trichoderma sp. cultures within only 13 days of incubation. The differential of the proposed methodology is the multiplication step in commercial organic substrates, which optimizes and reduces the production cost of the fungal inoculum, thus facilitating its use in the production of different species of seedlings of agricultural and forestry interest, as well as in research that aims to evaluate the agricultural potential of different Trichoderma sp. isolates.