Summary: Filamentous fungi are known as producers of a large array of diverse secondary metabolites (SMs) that aid in securing their environmental niche. Here, we demonstrated that the SMs have an additional role in fungal defence against other fungi: Trichoderma guizhouense , a mycoparasite, is able to antagonize Fusarium oxysporum f. sp. cubense race 4 (Foc4) by forming aerial hyphae that kill the host with hydrogen peroxide. At the same time, a gene cluster comprising two polyketide synthases is strongly expressed. Using functional genetics, we characterized this cluster and identified its products as azaphilones (termed as trigazaphilones). The trigazaphilones were found lacking of antifungal toxicity but exhibited high radical scavenging activities. The antioxidant property of trigazaphilones was in vivo functional under various tested conditions of oxidative stress. Thus, we conclude that the biosynthesis of trigazaphilones serves as a complementary antioxidant mechanism and defends T. guizhouense against the hydrogen peroxide that it produces to combat other fungi like Foc4.

Light is perceived by photoreceptors in fungi and further integrated into the stress-activated MAPK HOG pathway, and thereby potentially activates the expression of genes for stress responses. This indicates that the precise control of light conditions can likely improve the conidial yield and stress resistance to guarantee the low cost and long shelf life of Trichoderma-based biocontrol agents and biofertilizers. In this study, effects of wavelengths and intensities of light on conidial yield and stress tolerance to osmotic, oxidative and pH stresses in Trichoderma guizhouense were investigated. We found that 2 μmol photons/(m² × s) of blue light increased the conidial yield more than 1000 folds as compared to dark condition and simultaneously enhanced conidial stress resistance. The enhanced conidial stress resistance is probably due to the upregulated stress-related genes in blue light, which is under the control of the blue light receptor BLR1 and the MAP kinase HOG1.

Trichoderma is a widely recognized plant-growth-promoting fungus that has been extensively utilized in various agricultural applications. However, research on the economic production of Trichoderma spores and their effects on tea cuttings must be further advanced. In this study, T. guizhouense NJAU 4742 (NJAU 4742) emerged as a growth-promoting strain for tea cuttings, and the spore-production conditions of NJAU 4742 attained through solid-state fermentation (SSF) using tea residues were optimized. In a pot experiment, nursery substrates containing different concentrations of NJAU 4742 spores were tested for their influence on tea cutting growth and the rhizosphere fungal community. The optimal conditions for spore yield were determined as a 7:3 (w/w) ratio of tea residue to rice bran, a material thickness of 3 cm, an inoculum concentration of 15% (v/w), and an incubation time of 4 days, resulting in a spore count of 1.8 × 10⁹ CFU/g. Applying NJAU 4742 spore products significantly increased the biomass of tea cuttings and influenced the fungal community composition. Moreover, higher concentrations of NJAU 4742 spores yielded better growth performance, and applying nursery substrate with 1.0 × 10⁷ CFU/mL spores was the most economically viable option. Notably, among the top ten fungal genera with the highest relative abundance, Trichoderma showed a positive correlation with the fresh weight of tea cuttings, while the others exhibited a negative correlation. Overall, utilizing tea residue for SSF to produce NJAU 4742 was a feasible approach, and the application of NJAU 4742 spores enhanced the growth of tea cuttings by increasing the relative abundance of Trichoderma.

Fungi have evolved different strategies to cope with oxidative stress in competitive environments, however, the understanding of mechanisms that govern antioxidant response specificity and adequacy remains rudimentary. Trichoderma guizhouense NJAU 4742, a biocontrol fungus with strong viability, exhibits notable resistance to oxidative stress. This study characterizes a previously unidentified antioxidant-related Zn₂-Cys₆ transcription factor, TgZct4, in T. guizhouense based on genomic and transcriptomic profiles. Unlike the well-conserved transcription factors, TgZct4 has a positive role in the regulation of antioxidant response to both low and high concentrations of hydrogen peroxide. TgZct4 responds rapidly to oxidative stress, with its transcript levels showing a positive correlation with increasing hydrogen peroxide concentrations. In addition, TgZct4 is found to modulate T. guizhouense’s defense response by reprogramming antioxidant activity through classic antioxidant enzymes according to transcriptomic analysis. And the catalase TgCat1, regulated by TgZct4, confers protection against oxidative stress either independently or in conjunction with another TgZct4-dependent catalase TgCat2 in T. guizhouense. Overall, this work reveals TgZct4 as an important factor regulating the oxidative stress response in T. guizhouense, which provides valuable insights into fungal stress tolerance mechanisms and may contribute to the future genetic enhancement of this fungi for use as a biological control agent.