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Volume 159, Issue Pt_2

The autophagy gene , involved in the formation of the autophagosome, contributes to cell differentiation and growth but is dispensable for pathogenesis in the entomopathogenic fungus Free

Abstract

Autophagy is a highly conserved process, representing the major eukaryotic degradative pathway of cellular components. Autophagy-mediated recycling of cellular materials contributes to cell differentiation, tissue remodelling and proper development. In fungi, autophagy is required for normal growth and cell differentiation. The entomopathogenic fungus and its invertebrate targets represent a unique model system with which to examine host–pathogen interactions. The gene is one of 17 involved in autophagosome formation, and the homologue () was identified. The role of autophagy in growth and virulence was investigated via construction of a targeted gene knockout of . The mutant strain displayed increased sensitivity to nutrient limitation, with decreased germination and growth as compared with the wild-type parent. Conidiation was severely compromised and conidia derived from the Δ strain were altered in morphology. Cell differentiation into blastospores was also greatly reduced. Despite the significant growth and developmental defects, insect bioassays using the oriental leafworm moth, , indicated a modest (~40 %) decrease in virulence in the Δ strain. The phenotypic defects of the Δ strain could be restored by introduction of an intact copy of . These data suggest that unlike several plant and animal pathogenic fungi, where is required for infection, in it is dispensable for pathogenesis.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
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/content/journal/micro/10.1099/mic.0.062646-0
2013-02-01
2024-04-27
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References

  1. Bartoszewska M., Kiel J. A., Bovenberg R. A., Veenhuis M., van der Klei I. J. ( 2011). Autophagy deficiency promotes β-lactam production in Penicillium chrysogenum . Appl Environ Microbiol 77:1413–1422[PubMed] [CrossRef]
     [Google Scholar]
  2. Behie S. W., Zelisko P. M., Bidochka M. J. ( 2012). Endophytic insect-parasitic fungi translocate nitrogen directly from insects to plants. Science 336:1576–1577[PubMed] [CrossRef]
     [Google Scholar]
  3. Cho E.-M., Liu L., Farmerie W., Keyhani N. O. ( 2006). EST analysis of cDNA libraries from the entomopathogenic fungus Beauveria (Cordyceps) bassiana. I. Evidence for stage-specific gene expression in aerial conidia, in vitro blastospores and submerged conidia. Microbiology 152:2843–2854 [View Article][PubMed]
     [Google Scholar]
  4. Dong B., Liu X. H., Lu J. P., Zhang F. S., Gao H. M., Wang H. K., Lin F. C. ( 2009). MgAtg9 trafficking in Magnaporthe oryzae . Autophagy 5:946–954 [View Article][PubMed]
     [Google Scholar]
  5. Fan Y., Borovsky D., Hawkings C., Ortiz-Urquiza A., Keyhani N. O. ( 2012). Exploiting host molecules to augment mycoinsecticide virulence. Nat Biotechnol 30:35–37 [View Article][PubMed]
     [Google Scholar]
  6. Fang W. G., Feng J., Fan Y. H., Zhang Y. J., Bidochka M. J., Leger R. J., Pei Y. ( 2009). Expressing a fusion protein with protease and chitinase activities increases the virulence of the insect pathogen Beauveria bassiana . J Invertebr Pathol 102:155–159[PubMed] [CrossRef]
     [Google Scholar]
  7. Fang W. G., Vega-Rodríguez J., Ghosh A. K., Jacobs-Lorena M., Kang A., St Leger R. J. ( 2011). Development of transgenic fungi that kill human malaria parasites in mosquitoes. Science 331:1074–1077 [View Article][PubMed]
     [Google Scholar]
  8. Feng M. G., Poprawski T. J., Khachatourians G. G. ( 1994). Production, formulation and application of the entomopathogenic fungus Beauveria bassiana for insect control: current status. Biocontrol Sci Technol 4:3–34 [View Article]
     [Google Scholar]
  9. Gurulingappa P., Sword G. A., Murdoch G., McGee P. A. ( 2010). Colonization of crop plants by fungal entomopathogens and their effects on two insect pests when in planta. Biol Control 55:34–41 [View Article]
     [Google Scholar]
  10. Hanada T., Noda N. N., Satomi Y., Ichimura Y., Fujioka Y., Takao T., Inagaki F., Ohsumi Y. ( 2007). The Atg12–Atg5 conjugate has a novel E3-like activity for protein lipidation in autophagy. J Biol Chem 282:37298–37302[PubMed] [CrossRef]
     [Google Scholar]
  11. Holder D. J., Keyhani N. O. ( 2005). Adhesion of the entomopathogenic fungus Beauveria (Cordyceps) bassiana to substrata. Appl Environ Microbiol 71:5260–5266 [View Article][PubMed]
     [Google Scholar]
  12. Kikuma T., Kitamoto K. ( 2011). Analysis of autophagy in Aspergillus oryzae by disruption of Aoatg13, Aoatg4, and Aoatg15 genes. FEMS Microbiol Lett 316:61–69 [View Article][PubMed]
     [Google Scholar]
  13. Kirkland B. H., Westwood G. S., Keyhani N. O. ( 2004). Pathogenicity of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to Ixodidae tick species Dermacentor variabilis, Rhipicephalus sanguineus, and Ixodes scapularis . J Med Entomol 41:705–711 [View Article][PubMed]
     [Google Scholar]
  14. Klionsky D. J., Cuervo A. M., Seglen P. O. ( 2007). Methods for monitoring autophagy from yeast to human. Autophagy 3:181–206[PubMed] [CrossRef]
     [Google Scholar]
  15. Kuma A., Mizushima N., Ishihara N., Ohsumi Y. ( 2002). Formation of the approximately 350-kDa Apg12–Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast. J Biol Chem 277:18619–18625 [View Article][PubMed]
     [Google Scholar]
  16. Levine B., Klionsky D. J. ( 2004). Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell 6:463–477[PubMed] [CrossRef]
     [Google Scholar]
  17. Lewis M. W., Robalino I. V., Keyhani N. O. ( 2009). Uptake of the fluorescent probe FM4-64 by hyphae and haemolymph-derived in vivo hyphal bodies of the entomopathogenic fungus Beauveria bassiana . Microbiology 155:3110–3120 [View Article][PubMed]
     [Google Scholar]
  18. Liu X. H., Lu J. P., Zhang L., Dong B., Min H., Lin F. C. ( 2007). Involvement of a Magnaporthe grisea serine/threonine kinase gene, MgATG1, in appressorium turgor and pathogenesis. Eukaryot Cell 6:997–1005[PubMed] [CrossRef]
     [Google Scholar]
  19. Liu T. B., Liu X. H., Lu J. P., Zhang L., Min H., Lin F. C. ( 2010). The cysteine protease MoAtg4 interacts with MoAtg8 and is required for differentiation and pathogenesis in Magnaporthe oryzae . Autophagy 6:74–85 [View Article][PubMed]
     [Google Scholar]
  20. Liu X. H., Yang J., He R. L., Lu J. P., Zhang C. L., Lu S. L., Lin F. C. ( 2011). An autophagy gene, TrATG5, affects conidiospore differentiation in Trichoderma reesei . Res Microbiol 162:756–763 [View Article][PubMed]
     [Google Scholar]
  21. Lu J. P., Liu X. H., Feng X. X., Min H., Lin F. C. ( 2009). An autophagy gene, MgATG5, is required for cell differentiation and pathogenesis in Magnaporthe oryzae . Curr Genet 55:461–473 [View Article][PubMed]
     [Google Scholar]
  22. Michielse C. B., Hooykaas P. J. J., van den Hondel C. A. M. J. J., Ram A. F. J. ( 2008). Agrobacterium-mediated transformation of the filamentous fungus Aspergillus awamori . Nat Protoc 3:1671–1678[PubMed] [CrossRef]
     [Google Scholar]
  23. Nadal M., Gold S. E. ( 2010). The autophagy genes ATG8 and ATG1 affect morphogenesis and pathogenicity in Ustilago maydis . Mol Plant Pathol 11:463–478 [View Article][PubMed]
     [Google Scholar]
  24. Niemann A., Baltes J., Elsässer H.-P. ( 2001). Fluorescence properties and staining behavior of monodansylpentane, a structural homologue of the lysosomotropic agent monodansylcadaverine. J Histochem Cytochem 49:177–185[PubMed] [CrossRef]
     [Google Scholar]
  25. Park Y., Kim W., Kim A.-Y., Choi H. J., Choi J. K., Park N. S., Koh E. K., Seo J., Koh Y. H. ( 2011). Normal prion protein in Drosophila enhances the toxicity of pathogenic polyglutamine proteins and alters susceptibility to oxidative and autophagy signaling modulators. Biochem Biophys Res Commun 404:638–645[PubMed] [CrossRef]
     [Google Scholar]
  26. Pinan-Lucarré B., Balguerie A., Clavé C. ( 2005). Accelerated cell death in Podospora autophagy mutants. Eukaryot Cell 4:1765–1774[PubMed] [CrossRef]
     [Google Scholar]
  27. Pollack J. K., Harris S. D., Marten M. R. ( 2009). Autophagy in filamentous fungi. Fungal Genet Biol 46:1–8 [View Article][PubMed]
     [Google Scholar]
  28. Puttikamonkul S., Willger S. D., Grahl N., Perfect J. R., Movahed N., Bothner B., Park S., Paderu P., Perlin D. S., Cramer R. A. Jr ( 2010). Trehalose 6-phosphate phosphatase is required for cell wall integrity and fungal virulence but not trehalose biosynthesis in the human fungal pathogen Aspergillus fumigatus . Mol Microbiol 77:891–911
     [Google Scholar]
  29. Raeder U., Broda P. ( 1985). Rapid preparation of DNA from filamentous fungi. Lett Appl Microbiol 1:17–20 [View Article]
     [Google Scholar]
  30. Reumann S., Voitsekhovskaja O., Lillo C. ( 2010). From signal transduction to autophagy of plant cell organelles: lessons from yeast and mammals and plant-specific features. Protoplasma 247:233–256 [View Article][PubMed]
     [Google Scholar]
  31. Roetzer A., Gratz N., Kovarik P., Schüller C. ( 2010). Autophagy supports Candida glabrata survival during phagocytosis. Cell Microbiol 12:199–216[PubMed] [CrossRef]
     [Google Scholar]
  32. Suzuki K., Ohsumi Y. ( 2007). Molecular machinery of autophagosome formation in yeast, Saccharomyces cerevisiae . FEBS Lett 581:2156–2161 [View Article][PubMed]
     [Google Scholar]
  33. Tsukada M., Ohsumi Y. ( 1993). Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae . FEBS Lett 333:169–174[PubMed] [CrossRef]
     [Google Scholar]
  34. Veneault-Fourrey C., Barooah M., Egan M., Wakley G., Talbot N. J. ( 2006). Autophagic fungal cell death is necessary for infection by the rice blast fungus. Science 312:580–583 [View Article][PubMed]
     [Google Scholar]
  35. Wanchoo A., Lewis M. W., Keyhani N. O. ( 2009). Lectin mapping reveals stage-specific display of surface carbohydrates in in vitro and haemolymph-derived cells of the entomopathogenic fungus Beauveria bassiana . Microbiology 155:3121–3133 [View Article][PubMed]
     [Google Scholar]
  36. Xiao G., Ying S. H., Zheng P., Wang Z. L., Zhang S., Xie X.-Q., Shang Y., St Leger R. J., Zhao G. P. & other authors ( 2012). Genomic perspectives on the evolution of fungal entomopathogenicity in Beauveria bassiana . Sci Rep 2:483 [View Article][PubMed]
     [Google Scholar]
  37. Xie X. Q., Li F., Ying S. H., Feng M. G. ( 2012). Additive contributions of two manganese-cored superoxide dismutases (MnSODs) to antioxidation, UV tolerance and virulence of Beauveria bassiana . PLoS ONE 7:e30298 [View Article][PubMed]
     [Google Scholar]
  38. Ying S. H., Feng M. G. ( 2006). Novel blastospore-based transformation system for integration of phosphinothricin resistance and green fluorescence protein genes into Beauveria bassiana . Appl Microbiol Biotechnol 72:206–210 [View Article][PubMed]
     [Google Scholar]
  39. Ying S. H., Feng M. G. ( 2011). A conidial protein (CP15) of Beauveria bassiana contributes to the conidial tolerance of the entomopathogenic fungus to thermal and oxidative stresses. Appl Microbiol Biotechnol 90:1711–1720 [View Article][PubMed]
     [Google Scholar]
  40. Zhang S., Widemann E., Bernard G., Lesot A., Pinot F., Pedrini N., Keyhani N. O. ( 2012). CYP52X1, representing new cytochrome P450 subfamily, displays fatty acid hydroxylase activity and contributes to virulence and growth on insect cuticular substrates in entomopathogenic fungus Beauveria bassiana . J Biol Chem 287:13477–13486 [View Article][PubMed]
     [Google Scholar]
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The autophagy gene BbATG5, involved in the formation of the autophagosome, contributes to cell differentiation and growth but is dispensable for pathogenesis in the entomopathogenic fungus Beauveria bassiana
Microbiology 159, 243 (2013); https://doi.org/10.1099/mic.0.062646-0
/content/journal/micro/10.1099/mic.0.062646-0
/content/journal/micro/10.1099/mic.0.062646-0
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