Thornton, C. R. | bioRxiv Science

bioRxiv2025

Inducible volatile chemical signalling drives antifungal activity of Trichoderma hamatum GD12 during confrontation with the pathogen Sclerotinia sclerotiorum

BACKGROUNDThe use of beneficial soil fungi or their natural products offers a more sustainable alternative to synthetic fungicides for pathogen management in crops. Volatile organic compounds (VOCs) produced by such fungi act as semiochemicals that inhibit pathogens, with VOC production influenced by physical interactions between competing fungi. This study explores the interaction between the beneficial soil fungus Trichoderma hamatum GD12 strain (GD12), previously shown to antagonize crop pathogens such as Sclerotinia sclerotiorum, to test the hypothesis that its antagonistic effect is mediated by volatile chemical signalling. A GD12 mutant deficient in the chitinolytic enzyme N-acetyl-{beta}-glucosaminidase ({Delta}Thnag : : hph), which shows reduced biocontrol activity, was also examined. RESULTSIn dual-culture confrontation assays, co-inoculation of GD12 and S. sclerotiorum led to fungistatic interactions after 7 days, whereas{Delta} Thnag : : hph showed no antagonism, indicating a loss of antagonistic function. VOCs collected from individual and co-cultures were analysed by gas chromatography - flame ionization detector (GC-FID) analysis and coupled GC-mass spectrometry (GC-MS), revealing significant differences in VOC production between treatments, with VOC production notably upregulated in the GD12 + S. sclerotiorum co-culture. Peak production of 6-pentyl-2H-pyran-2-one occurred 17 days post-inoculation. This upregulation was absent in the{Delta} Thnag : : hph co-culture, suggesting VOCs may drive antagonism. Synthetic VOC assays revealed several compounds inhibitory to S. sclerotiorum, including 1-octen-3-one, which also arrested the growth of key fungal pathogens (Botrytis cinerea, Pyrenopeziza brassicae, and Gaeumannomyces tritici). Structural insights into 1-octen-3-ones antifungal activity against S. sclerotiorum are also presented. CONCLUSIONSThese findings support the hypothesis that the antagonistic properties of T. hamatum GD12 against crop fungal pathogens can, in part, be attributed to VOC production. Further research is needed to assess the potential of these semiochemicals as tools for pathogen management in agriculture.

microbiology↗

bioRxiv2019

3D light sheet fluorescence microscopy of lungs to dissect local host immune - Aspergillus fumigatus interactions

Aspergillus fumigatus is an opportunistic fungal pathogen that can cause life-threatening invasive lung infections in immunodeficient patients. The cellular and molecular processes of infection during onset, establishment and progression are highly complex and depend on both fungal attributes and the immune status of the host. Therefore, preclinical animal models are paramount to investigate and gain better insight into the infection process. Yet, despite their extensive use, commonly employed murine models of invasive pulmonary aspergillosis are not well understood due to analytical limitations. Here we present quantitative light sheet fluorescence microscopy (LSFM) to describe fungal growth and the local immune response in whole lungs at cellular resolution within its anatomical context. We analyzed three very common murine models of pulmonary aspergillosis based on immunosuppression with corticosteroids, chemotherapy-induced leukopenia or myeloablative irradiation. LSFM uncovered distinct architectures of fungal growth and degrees of tissue invasion in each model. Furthermore, LSFM revealed the spatial distribution, interaction and activation of two key immune cell populations in antifungal defense: alveolar macrophages and polymorphonuclear neutrophils. Interestingly, the patterns of fungal growth correlated with the detected effects of the immunosuppressive regimens on the local immune cell populations. Moreover, LSFM demonstrates that the commonly used intranasal route of spore administration did not result in the desired intra-alveolar deposition, as more than 60% of fungal growth occurred outside of the alveolar space. Hence, LSFM allows for more rigorous characterization of murine models of invasive pulmonary aspergillosis and pinpointing their strengths and limitations.\n\nIMPORTANCEThe use of animal models of infection is essential to advance our understanding of complex host-pathogen interactions that take place during Aspergillus fumigatus lung infections. As in the case of humans, mice need to be immunosuppressed to become susceptible to invasive pulmonary aspergillosis, the most serious infection caused by A. fumigatus. There are several immunosuppressive regimens that are routinely used to investigate fungal growth and/or immune responses in murine models of invasive pulmonary aspergillosis (IPA). However, the precise consequences that each immunosuppressive model has on the local immune populations and for fungal growth are not completely understood. Here we employed light sheet fluorescence microscopy (LSFM) to analyze whole lungs at cellular resolution, to pin down the scenario commonly used IPA models. Our results will be valuable to optimize and refine animal models to maximize their use in future research.\n\nVISUAL ABSTRACTQuantitative light sheet fluorescence microscopy to dissect local host-pathogen interactions in the lung after A. fumigatus airway infection.\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC=\"FIGDIR/small/661157v1_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (42K):\norg.highwire.dtl.DTLVardef@4baf31org.highwire.dtl.DTLVardef@dad3c3org.highwire.dtl.DTLVardef@84aab9org.highwire.dtl.DTLVardef@1e45c0d_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗