The production of eight phytohormones by Trichoderma species is described, as well as the 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase (ACCD) activity, which diverts the ethylene biosynthetic pathway in plants. The use of the Trichoderma strains T. virens T49, T. longibrachiatum T68, T. spirale T75 and T. harzianum T115 served to demonstrate the diverse production of the phytohormones gibberellins (GA) GA₁ and GA₄, abscisic acid (ABA), salicylic acid (SA), auxin (indole-3-acetic acid: IAA) and the cytokinins (CK) dihydrozeatin (DHZ), isopenteniladenine (iP) and trans-zeatin (tZ) in this genus. Such production is dependent on strain and/or culture medium. These four strains showed different degrees of wheat root colonization. Fresh and dry weights, conductance, H₂O₂ content and antioxidant activities such as superoxide dismutase, peroxidase and catalase were analyzed, under optimal irrigation and water stress conditions, on 30-days-old wheat plants treated with four-day-old Trichoderma cultures, obtained from potato dextrose broth (PDB) and PDB-tryptophan (Trp). The application of Trichoderma PDB cultures to wheat plants could be linked to the plants’ ability to adapt the antioxidant machinery and to tolerate water stress. Plants treated with PDB cultures of T49 and T115 had the significantly highest weights under water stress. Compared to controls, treatments with strains T68 and T75, with constrained GA₁ and GA₄ production, resulted in smaller plants regardless of fungal growth medium and irrigation regime.

The search for endophytic fungi in the roots of healthy wheat plants from a non-irrigation field trial allowed us to select 4 out of a total of 54 cultivable isolates belonging to the genus Trichoderma, identified as T. harzianum T136 and T139, T. simmonsii T137, and T. afroharzianum T138. In vitro assays against the phytopathogenic fungus Fusarium graminearum showed that the T. harzianum strains had the highest biocontrol potential and that T136 exhibited the highest cellulase and chitinase activities. Production patterns of eight phytohormones varied among the Trichoderma strains. All four, when applied alone or in combination, colonized roots of other wheat cultivars and promoted seed germination, tillering, and plant growth under optimal irrigation conditions in the greenhouse. Apart from T136, the endophytic Trichoderma strains showed plant protection capacity against drought as they activated the antioxidant enzyme machinery of the wheat plants. However, T. simmonsii T137 gave the best plant size and spike weight performance in water-stressed plants at the end of the crop. This trait correlated with significantly increased production of indole acetic acid and abscisic acid and increased 1-aminocyclopropane-1-carboxylic acid deaminase activity by T137. This study shows the potential of Trichoderma endophytes and that their success in agricultural systems requires careful selection of suitable strains.

Fusarium head blight (FHB), caused by Fusarium graminearum (Fg), is a major disease of wheat not only due to crop yield losses but also because of food safety concerns, since Fg produces toxic trichotecenes, such as deoxynivalenol (DON). Different Trichoderma strains have shown biocontrol efficacy against various Fusarium spp. in a wide variety of pathosystems. In this work, the efficacy of T. asperellum T25, T. harzianum T136 and T. simmonsii T137, was assessed against Fg ITEM 124 in in vitro tests, which included dual cultures, as well as cellulose and cellophane membrane assays. The three Trichoderma strains inhibited Fg growth to varying degrees. However, only T25 and T136 demonstrated control of FHB in wheat when applied to spikes. By quantitative real-time PCR (qPCR) we analysed the expression of eight plant defence-related marker genes in wheat spikes inoculated with Trichoderma, or not, and subsequently infected with Fg. Only wheat spikes pre-treated with T25 or T136 look to activate the salicylic acid-dependent defence, in response to pathogen infection. Expression of tri genes, involved in DON biosynthesis, was analysed by qPCR in dual-culture Trichoderma-Fg confrontations in two different media, and in the plant spikes. Confrontation results indicated that tri gene expression depends on the Trichoderma strain and the culture medium, but the three Trichoderma strains reduced the expression of tri5 in the plant. Results show that T. asperellum T25 reduced FHB disease index by more than 60 %, and was the most effective biocontrol agent, employing direct mechanisms to limit Fg growth and indirect mechanisms by priming local plant defences.

Woronin bodies are unique organelles in Pezizomycotina fungi that allow hyphae compartmentalization and prevent cytoplasmatic bleeding after mechanical injury. Several studies have related the peroxisomal protein HEX1, the major component of Woronin bodies with other biological processes such as hyphal growth, osmotic stress tolerance and pathogenicity. Trichoderma spp. are plant-beneficial multipurpose biological control agents, and proteomic and transcriptomic studies have shown that HEX1 and its corresponding gene are overrepresented when grown in the presence of fungal cell walls and plant polymers. To further investigate the involvement of hex1 in Trichoderma biology, we generated hex1 deletion transformants using the wheat endophytic strain T. simmonsii T137 as host. Results confirmed that hex1 gene is involved in the prevention of cytoplasmatic bleeding, and also has a role in fungal growth and biocontrol potential against phytopathogenic fungi and oomycetes. The involvement of hex1 in the fungal response to osmotic and oxidative stresses is conditioned by the type of stress and by the nutrient richness of the culture medium. The hex1 deletion also affected the interaction with wheat, but did not affect the plant protective effect of T137 against water stress. Overall, this study shows the implication of HEX1 in a wide range of biological processes necessary for T. simmonsii to deploy its abilities to be used as an agriculturally beneficial fungus.