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Plant recognition by Trichoderma harzianum elicits upregulation of a novel secondary metabolite cluster required for colonization

ABSTRACT: Plant recognition by Trichoderma harzianum elicits upregulation of a novel secondary metabolite cluster required for colonization

PROVIDER: PRJNA953408 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24107321.fastq.gz	Fastqsanger.gz
	SRR24107322.fastq.gz	Fastqsanger.gz
	SRR24107323.fastq.gz	Fastqsanger.gz
	SRR24107324.fastq.gz	Fastqsanger.gz
	SRR24107325.fastq.gz	Fastqsanger.gz

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Plant recognition by Trichoderma harzianum elicits upregulation of a novel secondary metabolite cluster required for colonization

Project description:The plant recognition specific PCA cluster mediates early chemical communication between plant and fungus, is required for colonization and it is likely responsible for the high potential of T. harzianum and closely related species for biocontrol applications.

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Trichoderma harzianum CECT 2413 expression in liquid basal medium or glucose-, chitin-, or tomato plant-containing media

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Peptidoglycan metabolite photoaffinity reporters reveal direct binding to intracellular pattern recognition receptors and Arf GTPases

Project description:Extensive structure-activity studies of iE-DAP and MDP have demonstrated their selective activation of NOD1- and NOD2-expressing cells, respectively. Nonetheless, the direct binding of iE-DAP and MDP to NOD1 and NOD2, respectively, as well as other proteins in mammalian cells has not been systematically investigated. To address the direct interaction of iE-DAP and MDP with NOD1 and NOD2 in living cells, we synthesized a series of peptidoglycan metabolite photoaffinity chemical reporters containing a diazirine for photocrosslinking in cells and an alkyne tag for bioorthogonal detection of covalently-labeled proteins. The analysis of these peptidoglycan metabolite photoaffinity reporters demonstrate that iE-DAP and MDP can directly bind to NOD1 and NOD2 in mammalian cells. The crosslinking of these peptidoglycan metabolite photoaffinity reporters only occur with active variants of NOD1 and NOD2. Beyond direct binding to NOD2, the active MDP photoaffinity reporter selectively crosslinked Arf GTPases in NOD2-expressing cells and did so most prominently with GTP-bound Arf6, revealing the Arf proteins as novel direct pattern recognition receptors. Additional labeling and co-precipitation experiments suggest that MDP induces a complex between NOD2 and GTP-Arf6. This study highlights the utility of peptidoglycan metabolite photoaffinity reporters for characterizing their direct binding to intracellular pattern recognition receptors and identifying unpredicted cellular cofactors. The applications of these microbial-specific metabolite reporters should enable the discovery of other receptors, transporters and regulatory enzymes in host cells and microbes.

2020-05-26 | PXD012350 | Pride

Trichoderma harzianum

Project description:Trichoderma harzianum Genome sequencing

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Regulation of Trichoderma harzianum gene expression and protein production in submerged cultures with inactivated oomycete mycelium

Project description:The breakdown of oomycete necromass is an important source of organic matter for composting. How Trichoderma harzianum, an important composting fungus, regulates gene expression and produces exo-proteins for degradation of oomycete necromass is poorly understood, especially related to cellulose, an important component of oomycete necromass. Complementary techniques of chemical compositional analysis, transcriptomics, exo-proteomics, enzymatic assays, and fungal genetics were used to analyze the degradation of inactivated oomycete mycelial powder – a surrogate for oomycete necromass. In total, 1,556 genes were upregulated and 212 exo-proteins were produced in T. harzianum oomycete mycelial powder cultures, and about 25% of the produced proteins showed corresponding gene upregulation. The enzymes detected, such as β-1,3-glucanases, and β-1,4-glucanases (cellulases), matched well with the composition of oomycete mycelial powder. Linkage compositional analysis showed that the mycelial powder contained ~ 60% 1,3 linkages and ~19% 1,4 linkages. The enzyme cocktail from the submerged cultures converted approximately one-third of the mycelial powder to glucose by in vitro assays. The conversion of the mycelial powder to glucose was not substantially reduced by deletion of the cellulolytic transcriptional activator XYR1. Deletion of XYR1 did decrease cellulase activity but only ~1% of mycelial powder-induced genes appeared to be XYR1-regulated. In conclusion, T. harzianum produces suitable enzyme cocktails for oomycete mycelial powder degradation, with β-1,3-glucanases likely playing a more important role than cellulases. T. harzianum cellulases may either be relatively unimportant for the degradation, or may not be co-activated alongside CAZymes degrading less recalcitrant parts of the mycelial powder.

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Growth, enzymatic, and transcriptomic analysis of xyr1 deletion reveals a major regulator of plant biomass-degrading enzymes in Trichoderma harzianum

Project description:The regulation of plant biomass degradation by fungi is critical to the carbon cycle, and applications in bioproducts and biocontrol. Trichoderma harzianum is an important plant biomass degrader, enzyme producer, and biocontrol agent, but few putative major transcriptional regulators have been deleted in this species. The T. harzianum ortholog of the transcriptional activator XYR1/XlnR/XLR-1 was deleted, and the mutant strains were analyzed by growth profiling, enzymatic activities, and transcriptomics on cellulose. From plate cultures, the Δxyr1 mutant had reduced growth on D-xylose, xylan, and cellulose, and from shake-flask cultures with cellulose, the Δxyr1 mutant had ~ 90% lower β-glucosidase activity, and no detectable β-xylosidase or cellulase activity. The comparison of the transcriptomes from 18 h shake-flask cultures on D-fructose, without a carbon source, and cellulose, showed major effects of XYR1 deletion whereby the Δxyr1 mutant on cellulose was transcriptionally most similar to the cultures without a carbon source. The cellulose induced 43 plant biomass-degrading CAZymes including xylanases as well as cellulases, and most of these had massively lower expression in the Δxyr1 mutant. Expression of a sub-set of carbon catabolic enzymes, other transcription factors, and sugar transporters was also lower in the Δxyr1 mutant on cellulose. In summary, T. harzianum XYR1 is the master regulator of cellulases and xylanases, as well as regulating carbon catabolic enzymes.

2023-12-26 | GSE252008 | GEO

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