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Abstract AimsThe American leaf spot, caused by Mycena citricolor, is an important disease of coffee (Coffea arabica), mostly in Central America. Currently, there are limited pathogen control alternatives that are environment friendly and economically accessible. The use of fungi isolated from the plant endomycobiota in their native habitats is on the rise because studies show their great potential for biological control. To begin to generate a green alternative to control M. citricolor, the objectives of the present study were to (i) collect, identify, screen (in vitro and in planta), and select endophytic fungi from wild Rubiaceae collected in old-growth forests of Costa Rica; (ii) confirm endophytic colonization in coffee plantlets; (iii) evaluate the effects of the endophytes on plantlet development; and (iv) corroborate the antagonistic ability in planta. Methods and resultsThrough in vitro and in planta antagonism assays, we found that out of the selected isolates (i.e. Daldinia eschscholzii GU11N, Nectria pseudotrichia GUHN1, Purpureocillium aff. lilacinum CT24, Sarocladium aff. kiliense CT25, Trichoderma rifaii CT5, T. aff. crassum G1C, T. aff. atroviride G7T, T. aff. strigosellum GU12, and Xylaria multiplex GU14T), Trichoderma spp. produced the highest growth inhibition percentages in vitro. Trichoderma isolates CT5 and G1C were then tested in planta using Coffea arabica cv. caturra plantlets. Endophytic colonization was verified, followed by in planta growth promotion and antagonism assays. ConclusionsResults show that Trichoderma isolates CT5 and G1C have potential for plant growth promotion and antagonism against Mycena citricolor, reducing incidence and severity, and preventing plant mortality. 

The transition from conventional to organic agriculture is often challenged by the adaptation of biological control agents to environments heavily exposed to agrochemical pollutants. We studied Trichoderma species isolated from living leaf tissues of wild Rubiacaeae (coffee family) plants to determine their fungicide tolerance and potential for bioremoval. First, we assessed the in vitro tolerance to fungicides of four Trichoderma isolates ( Trichoderma rifaii T1, T . aff. crassum T2, T . aff. atroviride T3, and T . aff. strigosellum T4) by placing mycelial plugs onto solid media supplemented with seven different systemic and non-systemic fungicides. After a week, most of the fungicides did not significantly inhibit the growth of the isolates, except in the case of cyproconazole, where the only isolate able to grow was T1; however, the colony morphology was affected by the presence of fungicides. Second, biological removal potential was established for selected isolates. For this experiment, the isolates T1, T2, and T4 were independently inoculated into liquid media with the fungicides azoxystrobin, chlorothalonil, cyproconazole, and trifloxystrobin. After 14 days of incubation, a removal of up to 89% was achieved for chlorothalonil, 46.4% for cyproconazole, and 33.1% for trifloxystrobin using viable biomass. In the case of azoxystrobin, the highest removal (82.2%) occurred by adsorption to fungal biomass. Ecotoxicological tests in Daphnia magna revealed that T1 has the highest removal potential, achieving significant elimination of every fungicide, while simultaneously detoxifying the aqueous matrix (except in the case of cyproconazole). Isolate T4 also exhibited an intermediate efficiency, while isolate T2 was unable to detoxify the matrix in most cases. The removal and detoxification of cyproconazole failed with all the isolates. These findings suggest that endosphere of wild plants could be an attractive guild to find new Trichoderma species with promising bioremediation capabilities. In addition, the results demonstrate that attention should be placed when combining certain types of agrochemicals with antagonistic fungi in Integrated Pest and Disease Management strategies or when transitioning to organic agriculture.