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Septation of Infectious Hyphae Is Critical for Appressoria Formation and Virulence in the Smut Fungus


Differentiation of hyphae into specialized infection structures, known as appressoria, is a common feature of plant pathogenic fungi that penetrate the plant cuticle. Appressorium formation in U. maydis is triggered by environmental signals but the molecular mechanism of this hyphal differentiation is largely unknown. Infectious hyphae grow on the leaf surface by inserting regularly spaced retraction septa at the distal end of the tip cell leaving empty sections of collapsed hyphae behind. Here we show that formation of retraction septa is critical for appressorium formation and virulence in U. maydis. We demonstrate that the diaphanous-related formin Drf1 is necessary for actomyosin ring formation during septation of infectious hyphae. Drf1 acts as an effector of a Cdc42 GTPase signaling module, which also consists of the Cdc42-specific guanine nucleotide exchange factor Don1 and the Ste20-like kinase Don3. Deletion of drf1, don1 or don3 abolished formation of retraction septa resulting in reduced virulence. Appressorium formation in these mutants was not completely blocked but infection structures were found only at the tip of short filaments indicating that retraction septa are necessary for appressorium formation in extended infectious hyphae. In addition, appressoria of drf1 mutants penetrated the plant tissue less frequently.


Vyšlo v časopise: Septation of Infectious Hyphae Is Critical for Appressoria Formation and Virulence in the Smut Fungus. PLoS Pathog 7(5): e32767. doi:10.1371/journal.ppat.1002044
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002044

Souhrn

Differentiation of hyphae into specialized infection structures, known as appressoria, is a common feature of plant pathogenic fungi that penetrate the plant cuticle. Appressorium formation in U. maydis is triggered by environmental signals but the molecular mechanism of this hyphal differentiation is largely unknown. Infectious hyphae grow on the leaf surface by inserting regularly spaced retraction septa at the distal end of the tip cell leaving empty sections of collapsed hyphae behind. Here we show that formation of retraction septa is critical for appressorium formation and virulence in U. maydis. We demonstrate that the diaphanous-related formin Drf1 is necessary for actomyosin ring formation during septation of infectious hyphae. Drf1 acts as an effector of a Cdc42 GTPase signaling module, which also consists of the Cdc42-specific guanine nucleotide exchange factor Don1 and the Ste20-like kinase Don3. Deletion of drf1, don1 or don3 abolished formation of retraction septa resulting in reduced virulence. Appressorium formation in these mutants was not completely blocked but infection structures were found only at the tip of short filaments indicating that retraction septa are necessary for appressorium formation in extended infectious hyphae. In addition, appressoria of drf1 mutants penetrated the plant tissue less frequently.


Zdroje

1. EmmettRWParberyDG 1975 Appressoria. Annu Rev Phytopathol 147 165

2. TuckerSLTalbotNJ 2001 Surface attachment and pre-penetration stage development by plant pathogenic fungi. Annu Rev Phytopathol 39 385 417

3. BechingerCGiebelKFSchnellMLeidererPDeisingHB 1999 Optical measurements of invasive forces exerted by appressoria of a plant pathogenic fungus. Science 285 1896 1899

4. DixonKPXuJRSmirnoffNTalbotNJ 1999 Independent signaling pathways regulate cellular turgor during hyperosmotic stress and appressorium-mediated plant infection by Magnaporthe grisea. Plant Cell 11 2045 2058

5. KumamotoCA 2008 Molecular mechanisms of mechanosensing and their roles in fungal contact sensing. Nat Rev Microbiol 6 667 673

6. WilsonRATalbotNJ 2009 Under pressure: investigating the biology of plant infection by Magnaporthe oryzae. Nat Rev Microbiol 7 185 195

7. LanverDMendoza-MendozaABrachmannAKahmannR 2010 Sho1 and Msb2-related proteins regulate appressorium development in the smut fungus Ustilago maydis. Plant Cell 22 2085 2101

8. BanuettF 1992 Ustilago maydis, the delightful blight. Trends Genet 8 174 180

9. KämperJKahmannRBölkerMMaLJBrefortT 2006 Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis. Nature 444 97 101

10. Garcia-MuseTSteinbergGPérez-MartinJ 2003 Pheromone-induced G2 arrest in the phytopathogenic fungus Ustilago maydis. Eukaryot Cell 2 494 500

11. SteinbergGSchliwaMLehmlerCBölkerMKahmannR 1998 Kinesin from the plant pathogenic fungus Ustilago maydis is involved in vacuole formation and cytoplasmic migration. J Cell Sci 111 2235 2246

12. Mendoza-MendozaABerndtPDjameiAWeiseCLinneU 2009 Physical-chemical plant-derived signals induce differentiation in Ustilago maydis. Mol Microbiol 71 895 911

13. SchirawskiJBöhnertHUSteinbergGSnetselaarKAdamikowaL 2005 Endoplasmic reticulum glucosidase II is required for pathogenicity of Ustilago maydis. Plant Cell 17 3532 3543

14. BanuettFHerskowitzI 1994 Morphological transitions in the life cycle of Ustilago maydis and their genetic control by the a and b loci. Exp Mycol 18 247 266

15. BölkerMUrbanMKahmannR 1992 The a mating type locus of U. maydis specifies cell signaling components. Cell 68 441 450

16. KahmannRBölkerM 1996 Self/nonself recognition in fungi: old mysteries and simple solutions. Cell 85 145 148

17. KämperJReichmannMRomeisTBölkerMKahmannR 1995 Multiallelic recognition: nonself-dependent dimerization of the bE and bW homeodomain proteins in Ustilago maydis. Cell 81 73 83

18. HeimelKSchererMSchulerDKämperJ 2010 The Ustilago maydis Clp1 protein orchestrates pheromone and b-dependent signaling pathways to coordinate the cell cycle and pathogenic development. Plant Cell 22 2908 2922

19. MahlertMLevelekiLHlubekASandrockBBölkerM 2006 Rac1 and Cdc42 regulate hyphal growth and cytokinesis in the dimorphic fungus Ustilago maydis. Mol Microbiol 59 567 578

20. HlubekASchinkKOMahlertMSandrockBBölkerM 2008 Selective activation by the guanine nucleotide exchange factor Don1 is a main determinant of Cdc42 signaling specificity in Ustilago maydis. Mol Microbiol 68 615 623

21. SandrockBBöhmerCBölkerM 2006 Dual function of the germinal centre kinase Don3 during mitosis and cytokinesis in Ustilago maydis. Mol Microbiol 62 655 666

22. WeinzierlGLevelekiLHasselAKostGWannerG 2002 Regulation of cell separation in the dimorphic fungus Ustilago maydis. Mol Microbiol 45 219 231

23. HiggsHN 2005 Formin proteins: a domain-based approach. Trends Biochem Sci 30 342 353

24. KovarDRHarrisESMahaffyRHiggsHNPollardTD 2006 Control of the assembly of ATP- and ADP-actin by formins and profilin. Cell 124 423 435

25. PruyneDEvangelistaMYangCBiEZigmondS 2002 Role of formins in actin assembly: nucleation and barbed-end association. Science 297 612 615

26. LiuWSatoAKhadkaDBhartiRDiazH 2008 Mechanism of activation of the Formin protein Daam1. Proc Natl Acad Sci U S A 105 210 215

27. SagotIRodalAAMoseleyJGoodeBLPellmanD 2002 An actin nucleation mechanism mediated by Bni1 and profilin. Nat Cell Biol 4 626 631

28. CastrillonDHWassermanSA 1994 Diaphanous is required for cytokinesis in Drosophila and shares domains of similarity with the products of the limb deformity gene. Development 120 3367 3377

29. ChalkiaDNikolaidisNMakalowskiWKleinJNeiM 2008 Origins and evolution of the formin multigene family that is involved in the formation of actin filaments. Mol Biol Evol 25 2717 2733

30. EvangelistaMZigmondSBooneC 2003 Formins: signaling effectors for assembly and polarization of actin filaments. J Cell Sci 116 2603 2611

31. LiFHiggsHN 2003 The mouse Formin mDia1 is a potent actin nucleation factor regulated by autoinhibition. Curr Biol 13 1335 1340

32. BrachmannAWeinzierlGKämperJKahmannR 2001 Identification of genes in the bW/bE regulatory cascade in Ustilago maydis. Mol Microbiol 42 1047 1063

33. DayPRAnagnostakisSL 1971 Corn smut dikaryon in culture. Nat New Biol 231 19 20

34. SnetselaarKMMimsCW 1992 Sporidial fusion and infection of maize seedlings by the smut fungus Ustilago maydis. Mycologia 84 193 203

35. SchmitzHPKaufmannAKöhliMLaissuePPPhilippsenP 2006 From function to shape: a novel role of a formin in morphogenesis of the fungus Ashbya gossypii. Mol Biol Cell 17 130 145

36. BöhmerCBöhmerMBölkerMSandrockB 2008 Cdc42 and the Ste20-like kinase Don3 act independently in triggering cytokinesis in Ustilago maydis. J Cell Sci 121 143 148

37. EvangelistaMBlundellKLongtineMSChowCJAdamesN 1997 Bni1p, a yeast formin linking cdc42p and the actin cytoskeleton during polarized morphogenesis. Science 276 118 122

38. BöhmerCRippCBölkerM 2009 The germinal centre kinase Don3 triggers the dynamic rearrangement of higher-order septin structures during cytokinesis in Ustilago maydis. Mol Microbiol 74 1484 1496

39. Alvarez-TabaresIPerez-MartinJ 2010 Septins from the phyotpathogenic fungus Ustilago maydis are required for proper morphogenesis but dispensable for virulence. PLoS One 5 e12933

40. SpelligTBottinAKahmannR 1996 Green fluorescent protein (GFP) as a new vital marker in the phytopathogenic fungus Ustilago maydis. Mol Gen Genet 252 503 509

41. SchererMHeimelKStarkeVKämperJ 2006 The Clp1 protein is required for clamp formation and pathogenic development of Ustilago maydis. Plant Cell 18 2388 2401

42. EvangelistaMPruyneDAmbergDCBooneCBretscherA 2002 Formins direct Arp2/3-independent actin filament assembly to polarize cell growth in yeast. Nat Cell Biol 4 260 269

43. KikyoMTanakaKKameiTOzakiKFujiwaraT 1999 An FH domain-containing Bnr1p is a multifunctional protein interacting with a variety of cytoskeletal proteins in Saccharomyces cerevisiae. Oncogene 18 7046 7054

44. PruyneDBretscherA 2000 Polarization of cell growth in yeast. The role of the cortical actin cytoskeleton. J Cell Sci 113 571 585

45. ImamuraHTanakaKHiharaTUmikawaMKameiT 1997 Bni1p and Bnr1p: downstream targets of the Rho family small G-proteins which interact with profilin and regulate actin cytoskeleton in Saccharomyces cerevisiae. EMBO J 16 2745 2755

46. KameiTTanakaKHiharaTUmikawaMImamuraH 1998 Interaction of Bnr1p with a novel Src homology 3 domain-containing Hof1p. Implication in cytokinesis in Saccharomyces cerevisiae. J Biol Chem 273 28341 28345

47. BatheMChangF 2010 Cytokinesis and the contractile ring in fission yeast: towards a systems-level understanding. Trends Microbiol 18 38 45

48. ChangFWoollardANurseP 1996 Isolation and characterization of fission yeast mutants defective in the assembly and placement of the contractile actin ring. J Cell Sci 109 131 142

49. ChangFDrubinDNurseP 1997 cdc12p, a protein required for cytokinesis in fission yeast, is a component of the cell division ring and interacts with profiling. J Cell Biol 137 169 182

50. YonetaniAChangF 2010 Regulation of Cytokinesis by the forming Cdc12p. Curr Biol 20 561 566

51. OlsonMF 2003 Dispatch. GTPase signaling: new functions for Diaphanous-related formins. Curr Biol 13 R360 362

52. SchirenbeckABretschneiderTArasadaRSchleicherMFaixJ 2005 The Diaphanous-related formin dDia2 is required for the formation and maintenance of filopodia. Nat Cell Biol 7 619 625

53. RobinowCF 1963 Observations on cell growth, mitosis, and division in the fungus Basidiobolus ranarum. J Cell Biol 17 123 152

54. IngoldCT 1982 Retraction-septa in various fungi. Trans Br Mycol Soc 79 373 38

55. de JongJCMcCormackBJSmirnoffNTalbotNJ 1997 Glycerol generates turgor in rice blast. Nature 389 244 245

56. ChoiWDeanRA 1997 The adenylate cyclase gene MAC1 of Magnaporthe grisea controls appressorium formation and other aspects of growth and development. Plant Cell 9 1973 1983

57. DöhlemannGWahlRVranesMde VriesRPKämperJ 2008 Establishment of compatibility in the Ustilago maydis/maize pathosystem. J Plant Physiol 165, 29-40

58. Veneault-FourreyCBarooahMEganMWakleyGTalbotNJ 2006 Autophagic fungal cell death is necessary for infection by the rice blast fungus. Science 312 580 583

59. KershawMJTalbotNJ 2009 Genome-wide functional analysis reveals that infection-assciated fungal autophagy is essential for rice blast disease. Proc Natl Acad Sci U S A 106 15967 15972

60. SaundersDGDagdasYFTalbotNJ 2010 Spatial uncoupling of mitosis and cytokinesis during appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae. Plant Cell 22 2417 2428

61. ChristensenJJ 1963 Corn smut caused by Ustilago maydis. Am. Phytopathol. Soc. Monogr. No 2

62. WadsworthP 2005 Cytokinesis: rho marks the spot. Curr Biol 15 R871 874

63. OliferenkoSChewTGBalasubramanianMK 2009 Positioning cytokinesis. Genes Dev 23 660 674

64. SchulzBBanuettFDahlMSchlesingerRSchäferW 1990 The b alleles of U. maydis, whose combinations program pathogenic development, code for polypeptides containing a homeodomain-related motif. Cell 60 295 306

65. LoubradouGBrachmannAFeldbrüggeMKahmannR 2001 A homologue of the transcriptional repressor Ssn6p antagonizes cAMP signaling in Ustilago maydis. Mol Microbiol 40 719 730

66. BrachmannAKönigJJuliusCFeldbrüggeM 2004 A reverse genetic approach for generating gene replacement mutants in Ustilago maydis. Mol Genet Genomics 272 216 226

67. BottinAKämperJKahmannR 1996 Isolation of a carbon source-regulated gene from Ustilago maydis. Mol Gen Genet 253 342 352

68. BöhmerMColbyTBöhmerCBräutigamASchmidtJ 2007 Proteomic analysis of dimorphic transition in the phytopathogenic fungus Ustilago maydis. Proteomics 7 675 685

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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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