1887

Abstract

Pyriculol was isolated from the rice blast fungus and found to induce lesion formation on rice leaves. These findings suggest that it could be involved in virulence. The gene was identified to encode a polyketide synthase essential for the production of the polyketide pyriculol in the rice blast fungus . The transcript abundance of correlates with the biosynthesis rate of pyriculol in a time-dependent manner. Furthermore, gene inactivation of resulted in a mutant unable to produce pyriculol, pyriculariol and their dihydro derivatives. Inactivation of a putative oxidase-encoding gene , which was found to be located in the genome close to , resulted in a mutant exclusively producing dihydropyriculol and dihydropyriculariol. By contrast, overexpression of resulted in a mutant strain only producing pyriculol. The cluster, furthermore, comprises two transcription factors and , which were both found individually to act as negative regulators repressing gene expression of . Additionally, extracts of and made from axenic cultures failed to induce lesions on rice leaves compared to extracts of the wild-type strain. Consequently, pyriculol and its isomer pyriculariol appear to be the only lesion-inducing secondary metabolites produced by wild-type (WT) under these culture conditions. Interestingly, the mutants unable to produce pyriculol and pyriculariol were as pathogenic as WT, demonstrating that pyriculol is not required for infection.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000396
2017-04-01
2019-11-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/163/4/541.html?itemId=/content/journal/micro/10.1099/mic.0.000396&mimeType=html&fmt=ahah

References

  1. Horbach R, Navarro-Quesada AR, Knogge W, Deising HB. When and how to kill a plant cell: infection strategies of plant pathogenic fungi. J Plant Physiol 2011;168:51–62 [CrossRef][PubMed]
    [Google Scholar]
  2. Dean R, van Kan JA, Pretorius ZA, Hammond-Kosack KE, di Pietro A et al. The top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol 2012;13:414–430 [CrossRef][PubMed]
    [Google Scholar]
  3. Yan X, Talbot NJ. Investigating the cell biology of plant infection by the rice blast fungus Magnaporthe oryzae. Curr Opin Microbiol 2016;34:147–153 [CrossRef][PubMed]
    [Google Scholar]
  4. Thines E. MAP kinase and protein kinase A-dependent mobilization of triacylglycerol and glycogen during appressorium turgor generation by Magnaporthe grisea. Plant Cell 2000;12:1703–1718
    [Google Scholar]
  5. Langfelder K, Streibel M, Jahn B, Haase G, Brakhage AA. Biosynthesis of fungal melanins and their importance for human pathogenic fungi. Fungal Genet Biol 2003;38:143–158 [CrossRef][PubMed]
    [Google Scholar]
  6. Howard RJ, Ferrari MA. Role of melanin in appressorium function. Exp Mycol 1989;13:403–418 [CrossRef]
    [Google Scholar]
  7. Collemare J, Billard A, Böhnert HU, Lebrun MH. Biosynthesis of secondary metabolites in the rice blast fungus Magnaporthe grisea: the role of hybrid PKS-NRPS in pathogenicity. Mycol Res 2008;112:207–215 [CrossRef][PubMed]
    [Google Scholar]
  8. Song Z, Bakeer W, Marshall JW, Yakasai AA, Khalid RM et al. Heterologous expression of the avirulence gene ACE1 from the fungal rice pathogen Magnaporthe oryzae. Chem Sci 2015;6:4837–4845 [CrossRef]
    [Google Scholar]
  9. Nukina M. The blast disease fungi and their metabolic products. J Pestic Sci 1999;24:293–298 [CrossRef]
    [Google Scholar]
  10. Tsurushima T, Minami Y, Miyagawa H, Nakayashiki H, Tosa Y et al. Induction of chlorosis, ROS generation and cell death by a toxin isolated from Pyricularia oryzae. Biosci Biotechnol Biochem 2010;74:2220–2225 [CrossRef]
    [Google Scholar]
  11. Tsurushima T, don LD, Kawashima K, Murakami J, Nakayashiki H et al. Pyrichalasin H production and pathogenicity of Digitaria-specific isolates of Pyricularia grisea. Mol Plant Pathol 2005;6:605–613 [CrossRef][PubMed]
    [Google Scholar]
  12. Umetsu N, Kaji J, Tamari K. Investigation on the toxin production by several blast fungus strains and isolation of tenuazonic acid as a novel toxin. Agric Biol Chem 1972;36:859–866 [CrossRef]
    [Google Scholar]
  13. Lebrun MH, Dutfoy F, Gaudemer F, Kunesch G, Gaudemer A. Detection and quantification of the fungal phytotoxin tenuazonic acid produced by Pyricularia oryzae. Phytochemistry 1990;29:3777–3783 [CrossRef]
    [Google Scholar]
  14. Yun CS, Motoyama T, Osada H. Biosynthesis of the mycotoxin tenuazonic acid by a fungal NRPS-PKS hybrid enzyme. Nat Commun 2015;6:8758 [CrossRef][PubMed]
    [Google Scholar]
  15. Hof C, Eisfeld K, Welzel K, Antelo L, Foster AJ et al. Ferricrocin synthesis in Magnaporthe grisea and its role in pathogenicity in rice. Mol Plant Pathol 2007;8:163–172 [CrossRef][PubMed]
    [Google Scholar]
  16. Patkar RN, Xue YK, Shui G, Wenk MR, Naqvi NI. Abc3-mediated efflux of an endogenous digoxin-like steroidal glycoside by Magnaporthe oryzae is necessary for host invasion during blast disease. PLoS Pathog 2012;8:e1002888 [CrossRef][PubMed]
    [Google Scholar]
  17. Parker D, Beckmann M, Zubair H, Enot DP, Caracuel-Rios Z et al. Metabolomic analysis reveals a common pattern of metabolic re-programming during invasion of three host plant species by Magnaporthe grisea. Plant J 2009;59:723–737 [CrossRef][PubMed]
    [Google Scholar]
  18. Tanaka K, Sasaki A, Cao H-Q, Yamada T, Igarashi M et al. Synthesis and biotransformation of plausible biosynthetic intermediates of salicylaldehyde-type phytotoxins of rice blast fungus, Magnaporthe grisea. European J Org Chem 2011;2011:6276–6280 [CrossRef]
    [Google Scholar]
  19. Jacob S, Foster AJ, Yemelin A, Thines E. Histidine kinases mediate differentiation, stress response, and pathogenicity in Magnaporthe oryzae. MicrobiologyOpen 2014;3:668–687 [CrossRef][PubMed]
    [Google Scholar]
  20. Green MR, Sambrook J. Molecular cloning: a laboratory manual, 4th ed. Cold Spring Harbor, NY: Cold Spring Harbor laboratory; 2012
    [Google Scholar]
  21. Odenbach D, Breth B, Thines E, Weber RW, Anke H et al. The transcription factor Con7p is a central regulator of infection-related morphogenesis in the rice blast fungus Magnaporthe grisea. Mol Microbiol 2007;64:293–307 [CrossRef][PubMed]
    [Google Scholar]
  22. Jacob S, Foster AJ, Yemelin A, Thines E. High osmolarity glycerol (HOG) signalling in Magnaporthe oryzae: identification of MoYPD1 and its role in osmoregulation, fungicide action, and pathogenicity. Fungal Biol 2015;119:580–594 [CrossRef][PubMed]
    [Google Scholar]
  23. Kramer B, Thines E, Foster AJ. MAP kinase signalling pathway components and targets conserved between the distantly related plant pathogenic fungi Mycosphaerella graminicola and Magnaporthe grisea. Fungal Genet Biol 2009;46:667–681 [CrossRef][PubMed]
    [Google Scholar]
  24. Tamura K, Peterson D, Peterson N, Stecher G, Nei M et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011;28:2731–2739 [CrossRef][PubMed]
    [Google Scholar]
  25. Larkin MA, Blackshields G, Brown NP, Chenna R, Mcgettigan PA et al. Clustal W and clustal X version 2.0. Bioinformatics 2007;23:2947–2948 [CrossRef][PubMed]
    [Google Scholar]
  26. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406–425[PubMed]
    [Google Scholar]
  27. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001;29:e45 [CrossRef][PubMed]
    [Google Scholar]
  28. Mattevi A, Fraaije MW, Mozzarelli A, Olivi L, Coda A et al. Crystal structures and inhibitor binding in the octameric flavoenzyme vanillyl-alcohol oxidase: the shape of the active-site cavity controls substrate specificity. Structure 1997;5:907–920 [CrossRef][PubMed]
    [Google Scholar]
  29. Kim Y, Park SY, Kim D, Choi J, Lee YH et al. Genome-scale analysis of ABC transporter genes and characterization of the ABCC type transporter genes in Magnaporthe oryzae. Genomics 2013;101:354–361 [CrossRef][PubMed]
    [Google Scholar]
  30. Markham JE, Hille J. Host-selective toxins as agents of cell death in plant–fungus interactions. Mol Plant Pathol 2001;2:229–239 [CrossRef][PubMed]
    [Google Scholar]
  31. Nukina M, Namai T. Productivity of pyrichalasin H, a phytotoxic metabolite, from different isolates of Pyricularia grisea and from other isolates of Pyricularia spp. Agric Biol Chem 1991;55:1899–1900 [CrossRef]
    [Google Scholar]
  32. Kim JC, Min JY, Kim HT, Cho KY, Yu SH. Pyricuol, a new phytotoxin from Magnaporthe grisea. Biosci Biotechnol Biochem 1998;62:173–174 [CrossRef][PubMed]
    [Google Scholar]
  33. Fischbach MA, Walsh CT. Assembly-line enzymology for polyketide and nonribosomal peptide antibiotics: logic, machinery, and mechanisms. Chem Rev 2006;106:3468–3496 [CrossRef][PubMed]
    [Google Scholar]
  34. Baker SE, Kroken S, Inderbitzin P, Asvarak T, Li B-Y et al. Two polyketide synthase-encoding genes are required for biosynthesis of the polyketide virulence factor, T-toxin, by Cochliobolus heterostrophus. Mol Plant Microbe Interact 2006;19:139–149[CrossRef]
    [Google Scholar]
  35. Khosla C, Gokhale RS, Jacobsen JR, Cane DE. Tolerance and specificity of polyketide synthases. Annu Rev Biochem 1999;68:219–253 [CrossRef][PubMed]
    [Google Scholar]
  36. Frandsen RJ, Nielsen NJ, Maolanon N, Sørensen JC, Olsson S et al. The biosynthetic pathway for aurofusarin in Fusarium graminearum reveals a close link between the naphthoquinones and naphthopyrones. Mol Microbiol 2006;61:1069–1080 [CrossRef][PubMed]
    [Google Scholar]
  37. Fujii I, Watanabe A, Sankawa U, Ebizuka Y. Identification of Claisen cyclase domain in fungal polyketide synthase WA, a naphthopyrone synthase of Aspergillus nidulans. Chem Biol 2001;8:189–197 [CrossRef][PubMed]
    [Google Scholar]
  38. Iwasaki S, Muro H, Sasaki K, Nozoe S, Okuda S et al. Isolations of phytotoxic substances produced by Pyricularia oryzae Cavara. Tetrahedron Lett 1973;14:3537–3542 [CrossRef]
    [Google Scholar]
  39. Manabu N, Takeshi S, Michimasa I, Takeshi U, Hisao T. Pyriculariol, a new phytotoxic metabolite of Pyricularia oryzae cavara. Agric Biol Chem 1981;45:2161–2162 [CrossRef]
    [Google Scholar]
  40. Iwasaki S, Nozoe S, Okuda S, Sato Z, Kozaka T. Isolation and structural elucidation of a phytotoxic substance produced by Pyricularia oryzae Cavara. Tetrahedron Lett 1969;10:3977–3980 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000396
Loading
/content/journal/micro/10.1099/mic.0.000396
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error