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Transcriptome insight into lignin utilization by Curvularia clavata and identification of regulators of laccase activity

ABSTRACT: Filamentous fungi are promising organisms for lignin degradation and mineralization. However, novel lignin-degrading fungal species are underexplored. Here, we isolated a fungal strain of Curvularia clavata that can utilize lignosulfonate as the carbon source and exhibited a relative high laccase activity during growth on lignosulfonate. Comparative transcriptomic analysis of the WT strain grown on glucose and lignosulfonate indicates that lignosulfonate and/or its metabolites have a significant effect on the gene expression profiles of C. clavata J1. Three regulators of laccase activity were identified, including a methyltransferase CcLaeA and two transcription factors, Rpn-4 and Tah-1. When grown on lignosulfonate, the laccase activity of the CclaeA and rpn-4 disrupted mutants (ΔCclaeA and Δrpn-4) increased by 49.2% and 43.5%, respectively, compared to the wild-type (WT) strain, whereas the tah-1 disrupted mutant (Δtah-1) decreased by 59.2%.

ORGANISM(S): Curvularia clavata

PROVIDER: GSE278592 | GEO | 2024/10/09

REPOSITORIES: GEO

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Project description:Secreted laccases are important enzymes on an ecological scale for their role in mediating plant-fungal interactions, but their function in fungal pathogenesis has yet to be elucidated. Ascomycete laccases have been primarily associagted with cell wall melanin deposition, and laccase mutants in ascomycete species often demonstrate reduced pigmentation. In this study, a putatively secreted laccase, Sslac2, was characterized from the broad host-range plant pathogen Sclerotinia sclerotiorum, which is largely unpigmented and is not dependent on melanogenesis for plant infection. Of the seven putative laccases in the S. sclerotiorum genome, Sslac2 was the only one found to be highly upregulated during pathogenesis of soybeans and was additionally found to be induced during growth on solid surfaces. Gene knockoues ot Sslac2 demonstrate wide ranging developmental defects, including abolished sclerotial formation, and are functionally non-pathogenic on unwounded tissue. While these mutants demonstrated a clear radial growth defect, enhanced growth was observed in liquid culture, likely due to altered hydrophobicity and thigmotropic responsiveness. Sslac2 mutants were also unable to respond to a host environment, and accordingly unable to differentiate penetration structures, respond appropriately to chemical stress, or produce the key virulence factor oxalic acid. Transmission and scanning electron microscopy of WT and mutant strains show apparent differences in extracellular matrix structure that may explain the inability of the mutant to respond to environmental cues. Targeting Sslac2 using host-induced gene silencing significantly improved resistance to S. sclerotiorum, suggesting that fungal laccases could be a valuable target of disease control. Collectively, we identified a laccase critical to the development and virulence of the broad host-range pathogen S. sclerotiorum and propose a potentially novel role for funal laccases in modulating environmental sensing.

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Transcriptome insight into lignin utilization by Curvularia clavata and identification of regulators of laccase activity

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