1887

Abstract

Mutants of the wheat pathogenic fungus lacking G-protein subunits display a variety of phenotypes including melanization defects, primary metabolic changes and a decreased ability to sporulate. To better understand the causes of these phenotypes, strains lacking a Gα, Gβ or Gγ subunit were compared to a wild-type strain using metabolomics. Agar plate growth at 22 °C revealed a number of fundamental metabolic changes and highlighted the influential role of these proteins in glucose utilization. A further characterization of the mutants was undertaken during prolonged storage at 4 °C, conditions known to induce sporulation in these sporulation-deficient signalling mutants. The abundance of several compounds positively correlated with the onset of sporulation including the dissacharide trehalose, the tryptophan degradation product tryptamine and the secondary metabolite alternariol; metabolites all previously associated with sporulation. Several other compounds decreased or were absent during sporulation. The levels of one such compound (Unknown_35.27_2194_319) decreased from being one of the more abundant compounds to absence during pycnidial maturation. This study has shed light on the role of G-protein subunits in primary metabolism during vegetative growth and exploited the cold-induced sporulation phenomenon in these mutants to identify some key metabolic changes that occur during asexual reproduction.

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2013-09-01
2019-12-11
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References

  1. Bailey A. , Mueller E. , Bowyer P. . ( 2000; ). Ornithine decarboxylase of Stagonospora (Septoria) nodorum is required for virulence toward wheat. . J Biol Chem 275:, 14242–14247. [CrossRef] [PubMed]
    [Google Scholar]
  2. Bringans S. , Hane J. K. , Casey T. , Tan K. C. , Lipscombe R. , Solomon P. S. , Oliver R. P. . ( 2009; ). Deep proteogenomics; high throughput gene validation by multidimensional liquid chromatography and mass spectrometry of proteins from the fungal wheat pathogen Stagonospora nodorum . . BMC Bioinformatics 10:, 301. [CrossRef] [PubMed]
    [Google Scholar]
  3. Casey T. , Solomon P. S. , Bringans S. , Tan K.-C. , Oliver R. P. , Lipscombe R. . ( 2010; ). Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification. . Proteomics 10:, 38–47. [CrossRef] [PubMed]
    [Google Scholar]
  4. Caspi R. , Foerster H. , Fulcher C. A. , Kaipa P. , Krummenacker M. , Latendresse M. , Paley S. , Rhee S. Y. , Shearer A. G. . & other authors ( 2008; ). The MetaCyc Database of metabolic pathways and enzymes and the BioCyc collection of pathway/genome databases. . Nucleic Acids Res 36: (Database issue), D623–D631. [CrossRef] [PubMed]
    [Google Scholar]
  5. Fang X. , Yano S. , Inoue H. , Sawayama S. . ( 2008; ). Lactose enhances cellulase production by the filamentous fungus Acremonium cellulolyticus . . J Biosci Bioeng 106:, 115–120. [CrossRef]
    [Google Scholar]
  6. Foreman P. K. , Brown D. , Dankmeyer L. , other authors . ( 2003; ). Transcriptional regulation of biomass-degrading enzymes in the filamentous fungus Trichoderma reesei . . J Biol Chem 278:, 31988–31997.[PubMed] [CrossRef]
    [Google Scholar]
  7. Gummer J. P. A. , Krill C. , Du Fall L. , Waters O. D. C. , Trengove R. D. , Oliver R. P. , Solomon P. S. . ( 2011; ). Metabolomics protocols for filamentous fungi. . In Plant Fungal Pathogens ( Methods in Molecular Biology , vol. 835), pp. 237–254. Edited by Bolton M. D. , Thomma B. P. H. J. . . New York:: Humana Press;.
    [Google Scholar]
  8. Gummer J. P. A. , Trengove R. D. , Oliver R. P. , Solomon P. S. . ( 2012; ). A comparative analysis of the heterotrimeric G-protein Gα, Gβ and Gγ subunits in the wheat pathogen Stagonospora nodorum . . BMC Microbiol 12:, 131. [CrossRef] [PubMed]
    [Google Scholar]
  9. IpCho S. V. S. , Tan K. C. , Koh G. , Gummer J. , Oliver R. P. , Trengove R. D. , Solomon P. S. . ( 2010; ). The transcription factor StuA regulates central carbon metabolism, mycotoxin production, and effector gene expression in the wheat pathogen Stagonospora nodorum . . Eukaryot Cell 9:, 1100–1108. [CrossRef] [PubMed]
    [Google Scholar]
  10. Jennings D. B. , Ehrenshaft M. , Pharr D. M. , Williamson J. D. . ( 1998; ). Roles for mannitol and mannitol dehydrogenase in active oxygen-mediated plant defense. . Proc Natl Acad Sci U S A 95:, 15129–15133. [CrossRef] [PubMed]
    [Google Scholar]
  11. Jennings D. B. , Daub M. E. , Pharr D. M. , Williamson J. D. . ( 2002; ). Constitutive expression of a celery mannitol dehydrogenase in tobacco enhances resistance to the mannitol-secreting fungal pathogen Alternaria alternata . . Plant J 32:, 41–49. [CrossRef] [PubMed]
    [Google Scholar]
  12. Kim J. D. , Kaiser K. , Larive C. K. , Borkovich K. A. . ( 2011; ). Use of 1H nuclear magnetic resonance to measure intracellular metabolite levels during growth and asexual sporulation in Neurospora crassa . . Eukaryot Cell 10:, 820–831. [CrossRef] [PubMed]
    [Google Scholar]
  13. Kraakman L. , Lemaire K. , Ma P. , Teunissen A. W. , Donaton M. C. V. , Van Dijck P. , Winderickx J. , de Winde J. H. , Thevelein J. M. . ( 1999; ). A Saccharomyces cerevisiae G-protein coupled receptor, Gpr1, is specifically required for glucose activation of the cAMP pathway during the transition to growth on glucose. . Mol Microbiol 32:, 1002–1012. [CrossRef] [PubMed]
    [Google Scholar]
  14. Lafon A. , Seo J. A. , Han K. H. , Yu J. H. , d’Enfert C. . ( 2005; ). The heterotrimeric G-protein GanB(α)-SfaD(β)-GpgA(γ) is a carbon source sensor involved in early cAMP-dependent germination in Aspergillus nidulans . . Genetics 171:, 71–80. [CrossRef] [PubMed]
    [Google Scholar]
  15. Li L. , Borkovich K. A. . ( 2006; ). GPR-4 is a predicted G-protein-coupled receptor required for carbon source-dependent asexual growth and development in Neurospora crassa . . Eukaryot Cell 5:, 1287–1300. [CrossRef] [PubMed]
    [Google Scholar]
  16. Lowe R. G. , Lord M. , Rybak K. , Trengove R. D. , Oliver R. P. , Solomon P. S. . ( 2008; ). A metabolomic approach to dissecting osmotic stress in the wheat pathogen Stagonospora nodorum . . Fungal Genet Biol 45:, 1479–1486. [CrossRef] [PubMed]
    [Google Scholar]
  17. Lowe R. G. T. , Lord M. , Rybak K. , Trengove R. D. , Oliver R. P. , Solomon P. S. . ( 2009; ). Trehalose biosynthesis is involved in sporulation of Stagonospora nodorum . . Fungal Genet Biol 46:, 381–389. [CrossRef] [PubMed]
    [Google Scholar]
  18. Oliver R. P. , Solomon P. S. . ( 2010; ). New developments in pathogenicity and virulence of necrotrophs. . Curr Opin Plant Biol 13:, 415–419. [CrossRef] [PubMed]
    [Google Scholar]
  19. Oliver R. P. , Friesen T. L. , Faris J. D. , Solomon P. S. . ( 2012; ). Stagonospora nodorum: from pathology to genomics and host resistance. . Annu Rev Phytopathol 50:, 23–43. [CrossRef] [PubMed]
    [Google Scholar]
  20. Seiboth B. , Pakdaman B. S. , Hartl L. , Kubicek C. P. . ( 2007; ). Lactose metabolism in filamentous fungi: how to deal with an unknown substrate. . Fungal Biol Rev 21:, 42–48. [CrossRef]
    [Google Scholar]
  21. Solomon P. S. , Tan K. C. , Sanchez P. , Cooper R. M. , Oliver R. P. . ( 2004; ). The disruption of a Gα subunit sheds new light on the pathogenicity of Stagonospora nodorum on wheat. . Mol Plant Microbe Interact 17:, 456–466. [CrossRef] [PubMed]
    [Google Scholar]
  22. Solomon P. S. , Tan K. C. , Oliver R. P. . ( 2005; ). Mannitol 1-phosphate metabolism is required for sporulation in planta of the wheat pathogen Stagonospora nodorum . . Mol Plant Microbe Interact 18:, 110–115. [CrossRef] [PubMed]
    [Google Scholar]
  23. Solomon P. S. , Lowe R. G. T. , Tan K. C. , Waters O. D. C. , Oliver R. P. . ( 2006a; ). Stagonospora nodorum: cause of stagonospora nodorum blotch of wheat. . Mol Plant Pathol 7:, 147–156. [CrossRef] [PubMed]
    [Google Scholar]
  24. Solomon P. S. , Rybak K. , Trengove R. D. , Oliver R. P. . ( 2006b; ). Investigating the role of calcium/calmodulin-dependent protein kinases in Stagonospora nodorum . . Mol Microbiol 62:, 367–381. [CrossRef] [PubMed]
    [Google Scholar]
  25. Solomon P. S. , Waters O. D. C. , Jörgens C. I. , Lowe R. G. T. , Rechberger J. , Trengove R. D. , Oliver R. P. . ( 2006c; ). Mannitol is required for asexual sporulation in the wheat pathogen Stagonospora nodorum (glume blotch). . Biochem J 399:, 231–239. [CrossRef] [PubMed]
    [Google Scholar]
  26. Solomon P. S. , Waters O. D. C. , Oliver R. P. . ( 2007; ). Decoding the mannitol enigma in filamentous fungi. . Trends Microbiol 15:, 257–262. [CrossRef] [PubMed]
    [Google Scholar]
  27. Tan K. C. , Heazlewood J. L. , Millar A. H. , Thomson G. , Oliver R. P. , Solomon P. S. . ( 2008; ). A signaling-regulated, short-chain dehydrogenase of Stagonospora nodorum regulates asexual development. . Eukaryot Cell 7:, 1916–1929. [CrossRef] [PubMed]
    [Google Scholar]
  28. Tan K. C. , Heazlewood J. L. , Millar A. H. , Oliver R. P. , Solomon P. S. . ( 2009a; ). Proteomic identification of extracellular proteins regulated by the Gna1 Gα subunit in Stagonospora nodorum . . Mycol Res 113:, 523–531. [CrossRef] [PubMed]
    [Google Scholar]
  29. Tan K. C. , Trengove R. D. , Maker G. L. , Oliver R. P. , Solomon P. S. . ( 2009b; ). Metabolite profiling identifies the mycotoxin alternariol in the pathogen Stagonospora nodorum . . Metabolomics 5:, 330–335. [CrossRef]
    [Google Scholar]
  30. Wilson R. A. , Jenkinson J. M. , Gibson R. P. , Littlechild J. A. , Wang Z. Y. , Talbot N. J. . ( 2007; ). Tps1 regulates the pentose phosphate pathway, nitrogen metabolism and fungal virulence. . EMBO J 26:, 3673–3685. [CrossRef] [PubMed]
    [Google Scholar]
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