Inhibition of Mycotoxigenic Fungi in Different Vegetable Matrices by Extracts of Trichoderma Species.

Post-harvest fungal diseases of plant products are a serious concern leading to economic losses and health risks. Moreover, the use of synthetic chemical fungicides to prevent these diseases is limited due to toxic residues. This study aimed at determining the effective dose of extracts of Trichoderma asperellum IMI393899 (TE1) and Trichoderma atroviride TS (TE2) in inhibiting the contamination by mycotoxigenic fungi on different plant matrices. Extracts were tested on tomatoes contaminated by Fusarium verticillioides and Fusarium graminearum, wheat contaminated by Penicillium verrucosum and maize contaminated by Aspergillus flavus. The efficacy of extracts was evaluated at two time intervals after treatment, 4 and 11 days for tomato, and 10 and 20 days for both wheat and maize. Both extracts showed a significant inhibitory activity on mycotoxigenic pathogens and significantly reduced Log CFU/g compared to the control. Moreover, the extracts reduced mycotoxin production in a dose dependent manner and with a long-lasting effect. The ochratoxin A was reduced by both extracts but only the extract TE2 was effective in reducing aflatoxins, whereas TE1 treatment increased their synthesis.

Antifungal Activity of Bioactive Metabolites Produced by Trichoderma asperellum and Trichoderma atroviride in Liquid Medium.

Trichoderma spp. are known as biocontrol agents of fungal plant pathogens and have been recognized as a potential source of bioactive metabolites. The production of antimicrobial substances from strains T. atroviride (TS) and T. asperellum (IMI 393899) was investigated. The bioactivity of 10- and 30-day culture filtrate extracted with ethyl acetate was assessed against a set of pathogenic fungi and oomycetes. The 30-day extracts of both strains had significant cytotoxic effects against the tested pathogens, with values of minimum fungicidal concentration (MFC) ranging between 0.19 and 6.25 mg/mL. Dual culture assay (direct contact and nondirect contact) and the percentage inhibition of radial growth (PIRG) was calculated. The highest PIRG values were 76% and 81% (direct contact) with IMI 393899 and TS, respectively. Nondirect contact does not show inhibition on any of pathogens tested, indicating that the inhibition is not due to the secretion of volatile substances. Culture filtrates were analyzed by GC-MS and HPLC-Q-TOF-MS for the identification of volatile organic compounds (VOCs) and nonvolatile organic compounds (nVOCs), respectively. Seven classes of VOCs and 12 molecules of nVOCs were identified. These results indicate that these strains of Trichoderma had antimicrobial activities and they are potential natural sources of compounds with biological activity.

Trichoderma Counteracts the Challenge of Phytophthora nicotianae Infections on Tomato by Modulating Plant Defense Mechanisms and the Expression of Crinkler, Necrosis-Inducing Phytophthora Protein 1, and Cellulose-Binding Elicitor Lectin Pathogenic Effectors.

Decoding the mechanisms of plant defense against plant pathogens in a scenario where antagonistic activity and the plant growth-promoting effects of useful organisms intervene simultaneously is a new frontier of plant pathology. Here, we demonstrated that (i) two selected strains of Trichoderma asperellum and Trichoderma atroviride promoted tomato (Solanum lycopersicum) growth and reduced the severity of disease caused by the oomycete Phytophthora nicotianae and (ii) the genetic patterns of the components of the experimental model system tomato-Trichoderma spp.-P. nicotianae were differentially expressed. The beneficial effects in both the promotion of the growth of host plant and the biological control of the pathogen by two selected strains of different Trichoderma species were tested both in planta and in vitro. In both respects, T. atroviride demonstrated to be more effective than T. asperellum. Additionally, the simultaneous transcriptional reprogramming of several plant defense-related genes, pathogen effectors, and mycoparasitism-related genes in tomato, P. nicotianae, and Trichoderma spp., respectively, was evaluated during the three-component interaction. Results support the hypothesis that Trichoderma spp. elicit the expression of plant defense-related genes. As expected, a mycoparasitism-related gene was significantly up-regulated in Trichoderma-colonizing tomato plants infected by P. nicotianae. Finally, a marked up-regulation of the genes encoding two necrosis-inducing effectors was observed in P. nicotianae infecting tomato plants colonized by Trichoderma. In conclusion, this study is a contribution toward understanding the genetic pathways related with the ability of Trichoderma spp. to counteract the challenge of P. nicotianae infections on tomato. Additionally, the experiments revealed the beneficial effects in the tomato growth promotion of a new T. atroviride strain and its good antagonistic effectiveness in the biological control of root and crown rot incited by P. nicotianae, confirming that Trichoderma spp. can be a powerful tool in integrated pest management strategies of Phytophthora diseases of horticultural crops.