Wheat head blight caused by Fusarium graminearum is one of the major wheat diseases in the world; therefore, it is very significant to develop an effective and environmentally friendly microbial fungicide against it. Trichoderma atroviride and Bacillus subtilis are widely applied biocontrol microorganisms with separate advantages; however, little work has been conducted for synergistically elevating the effects of biocontrol and plant promotion through the co-cultivation of the two microorganisms. Our study demonstrated that T. atroviride SG3403 is compatible with B. subtilis 22. The co-culture metabolites contained a group of antagonistic compounds which were able to inhibit F. graminearum growth and increase the activities of pathogen G protein and mitogen-activated protein kinase (MAPK) as compared with axenic culture metabolites. Additionally, the co-culture metabolites enabled us to more significantly decrease the production of gibberellin (GA), deoxynivalenol (DON), and zearalenone (ZEN) from F. graminearum, which disorganized the subcellular structure, particularly the cytoplasm of F. graminearum hyphae, relative to the axenically cultured metabolites. Furthermore, the seed-coating agent made by the co-culture had significant effects against F. graminearum infection by triggering the expression of host plant defensive genes, including PR1, PR3, PR4, PR5, ACS, and SOD. It is suggested that jasmonic acid and ethylene (JA/ET) signaling might dominate wheat’s induced systemic resistance (ISR) against wheat head blight. A dry, powdered bio-seed coating agent containing the co-culture mixtures was confirmed to be a bioavailable formulation that can be applied to control wheat head blight. Taken together, the co-culture’s metabolites or the metabolites and living cells might provide a basis for the further development of a new kind of microbial bio-fungicide in the future.

Trichoderma, a well-known and extensively studied fungal genus, has gained significant attention for its remarkable antagonistic abilities against a wide range of plant pathogens. In this study, a total of 108 Trichoderma isolates were screened through in vitro dual antagonistic assays and culture filtrate inhibition against Fusarium graminearum. Of these, the YNQJ1002 displayed noteworthy inhibitory activities along with thermal stability. To validate the metabolic differences between YNQJ1002 and GZLX3001 (with strong and weak antagonism, respectively), UPLC-TOF-MS/MS mass spectrometry was employed to analyze and compare the metabolite profiles. We identified 12 significantly up-regulated metabolites in YNQJ1002, which include compounds like Trigoneoside, Torvoside, trans,trans-hepta-2,4,6-trienoic acid, and Chamazulene. These metabolites are known for their antimicrobial properties or signaling roles as components of cell membranes. Enriched KEGG analysis revealed a significant enrichment in sphingolipid metabolism and linoleic acid metabolism, as well as autophagy. The results demonstrated that YNQJ1002’s abundance of antimicrobial substances, resulting from specific metabolic pathways, enhanced its superior antagonistic activity against F. graminearum. Finally, YNQJ1002 was identified using the ITS, tef1-1α, and rpb2 regions, with MIST system sequence matching confirming its classification within the species. Overall, we have obtained a novel strain, T. asperellum YNQJ1002, which is rich in metabolites and shows potential antagonistic activity against F. graminearum. This study has opened promising prospects for the development of innovative Trichoderma-derived antifungal compounds, featuring a unique mechanism against pathogens.

MicroRNAs (miRNAs) are known to play important roles in the immune response to pathogen infection in different plants. Further, Trichoderma strains are able to activate plant defense responses against pathogen attacks. However, little is known about the involvement of miRNAs in the defense response primed by Trichoderma strains. To explore the miRNAs sensitive to priming by Trichoderma, we studied the small RNAs and transcriptome changes in maize leaves that were systemically induced by seed treatment with Trichoderma harzianum (strain T28) against Cochliobolus heterostrophus (C. heterostrophus) infection in leaves. Through analysis of the sequencing data, 38 differentially expressed miRNAs (DEMs) and 824 differentially expressed genes (DEGs) were identified. GO and KEGG analyses of DEGs demonstrated that genes involved in the plant hormone signal transduction pathway and oxidation-reduction process were significantly enriched. In addition, 15 miRNA–mRNA interaction pairs were identified through the combined analysis of DEMs and DEGs. These pairs were supposed to play roles in the maize resistance primed by T. harzianum T28 to C. heterostrophus, in which miR390, miR169j, miR408b, miR395a/p, and novel miRNA (miRn5231) were more involved in the induction of maize resistance. This study provided valuable information for understanding the regulatory role of miRNA in the T. harzianum primed defense response.

Trichoderma spp. are widely used beneficial microbes in agricultural production; however, the improper carrier choice for Trichoderma agent preparation can alter the effectiveness of Trichoderma fungicides. In this study, the co-culture of four Trichoderma strains produced a large amount of free amino acids, with a content of 392.8414 ug/mL, and significantly improved the production level of γ-aminobutyric acid. A greenhouse experiment further showed that the co-culture of Trichoderma synergistically improved the female flower development and bacterial angular leaf spot resistance. The effects of ten kinds of carriers were compared in terms of water absorption and heat generation, as well as their effects on the seedling emergence rate and the plant growth promotion of maize, cucumber, and pakchoi cabbage. Each carrier was screened to mix with four strains of co-culture metabolites to prepare highly soluble and quality powders. The results showed that there were different effects of the carriers themselves and Trichoderma strain co-culture metabolite powder prepared with the carriers on seedling emergence rate and seedling growth. Β-cyclodextrin performed best in high solubility and low heat generation upon absorbing water and in easy drying in processing operations. Trichoderma strains co-culture metabolite powder with β-cyclodextrin as a carrier provided the most obvious promotion effects on seedling emergence rate and seedling growth. Therefore, β-cyclodextrin was determined to be an ideal carrier to prepare a highly water-soluble Trichoderma agent. Taken together, the study successfully developed a new type of highly soluble powder containing Trichoderma co-culture metabolites that is expected to benefit farming drip irrigation and spraying systems for the promotion of crop growth and disease control.

The putative methyltransferase Lae1 is a global regulator in Trichoderma, which modulates the expression of secondary metabolite gene clusters, possibly via chromatin remodeling. Here we aimed to explore the specific transcription and metabolites profiles regulated by Lae1 in T. atroviride 23. Comparative transcriptomics and metabolome analyses between the lae1 deletion (Mlae1) and over-expressing (Olae1) mutants were performed using RNA sequencing and QTOF-UPLC-MS techniques. In total, 1344 unique differentially expressed genes (DEGs) and 92 metabolites were identified across three strains. The significantly altered metabolic profiles revealed that the lae1 gene modulates central carbon metabolism, amino acid metabolism, secondary metabolism, and phospholipid metabolism. The effects of lae1 on phospholipid metabolism were further explored, and the findings showed that lae1 modulates the composition and function of cell membranes and other metabolic activities, including the phosphotransferase system (PTS) and biosynthesis of secondary metabolites (SM). Phospholipid metabolism is related to energy metabolism, signal transduction, and environmental adaptability of microorganisms. These data showed that Lae1 affects the primary metabolites, phospholipid, as well as the regulation of secondary metabolites in Trichoderma. This study could potentially provoke in-depth investigations of the Lae1-mediated target genes in phospholipid synthesis. The Lae1 may act as a novel target that is associated with disease defense and drug development in the future.

Curvularia lunata (Wakkre) Boedijn is an important pathogenic fungus that causes maize leaf spot, a prevalent disease that caused significant yield losses in maize-growing areas in China in the 1990s. Clpks18, a polyketide synthase (CLPKS18) gene, has been identified as a crucial virulence-related gene in C. lunata. However, the impact of Clpks18 and its biosynthesized virulence factor (R)-(-)-mellein on the expression of maize genes related to the defense signal pathway has never been determined. In this study, it was found that Clpks18 and (R)-(-)-mellein significantly interfere with the signaling pathways of JA and IAA in maize leaves but in different ways and in a time-dependent manner. While CLPKS18 inhibited the maize’s JA and IAA signaling pathways through its related secondary metabolite, (R)-(-)-mellein inhibited the JA signaling pathway but stimulated IAA accumulation in maize leaves. In summary, understanding this novel virulence effector’s mechanism of interference with maize resistance enriches the pathology of Curvularia leaf spot in maize on the one hand and provides a foundation for screening the resistance germplasm and chemical fungicides against the disease on the other.