#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Two Novel Transcriptional Regulators Are Essential for Infection-related Morphogenesis and Pathogenicity of the Rice Blast Fungus


The cyclic AMP-dependent protein kinase A signaling pathway plays a major role in regulating plant infection by the rice blast fungus Magnaporthe oryzae. Here, we report the identification of two novel genes, MoSOM1 and MoCDTF1, which were discovered in an insertional mutagenesis screen for non-pathogenic mutants of M. oryzae. MoSOM1 or MoCDTF1 are both necessary for development of spores and appressoria by M. oryzae and play roles in cell wall differentiation, regulating melanin pigmentation and cell surface hydrophobicity during spore formation. MoSom1 strongly interacts with MoStu1 (Mstu1), an APSES transcription factor protein, and with MoCdtf1, while also interacting more weakly with the catalytic subunit of protein kinase A (CpkA) in yeast two hybrid assays. Furthermore, the expression levels of MoSOM1 and MoCDTF1 were significantly reduced in both Δmac1 and ΔcpkA mutants, consistent with regulation by the cAMP/PKA signaling pathway. MoSom1-GFP and MoCdtf1-GFP fusion proteins localized to the nucleus of fungal cells. Site-directed mutagenesis confirmed that nuclear localization signal sequences in MoSom1 and MoCdtf1 are essential for their sub-cellular localization and biological functions. Transcriptional profiling revealed major changes in gene expression associated with loss of MoSOM1 during infection-related development. We conclude that MoSom1 and MoCdtf1 functions downstream of the cAMP/PKA signaling pathway and are novel transcriptional regulators associated with cellular differentiation during plant infection by the rice blast fungus.


Vyšlo v časopise: Two Novel Transcriptional Regulators Are Essential for Infection-related Morphogenesis and Pathogenicity of the Rice Blast Fungus. PLoS Pathog 7(12): e32767. doi:10.1371/journal.ppat.1002385
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002385

Souhrn

The cyclic AMP-dependent protein kinase A signaling pathway plays a major role in regulating plant infection by the rice blast fungus Magnaporthe oryzae. Here, we report the identification of two novel genes, MoSOM1 and MoCDTF1, which were discovered in an insertional mutagenesis screen for non-pathogenic mutants of M. oryzae. MoSOM1 or MoCDTF1 are both necessary for development of spores and appressoria by M. oryzae and play roles in cell wall differentiation, regulating melanin pigmentation and cell surface hydrophobicity during spore formation. MoSom1 strongly interacts with MoStu1 (Mstu1), an APSES transcription factor protein, and with MoCdtf1, while also interacting more weakly with the catalytic subunit of protein kinase A (CpkA) in yeast two hybrid assays. Furthermore, the expression levels of MoSOM1 and MoCDTF1 were significantly reduced in both Δmac1 and ΔcpkA mutants, consistent with regulation by the cAMP/PKA signaling pathway. MoSom1-GFP and MoCdtf1-GFP fusion proteins localized to the nucleus of fungal cells. Site-directed mutagenesis confirmed that nuclear localization signal sequences in MoSom1 and MoCdtf1 are essential for their sub-cellular localization and biological functions. Transcriptional profiling revealed major changes in gene expression associated with loss of MoSOM1 during infection-related development. We conclude that MoSom1 and MoCdtf1 functions downstream of the cAMP/PKA signaling pathway and are novel transcriptional regulators associated with cellular differentiation during plant infection by the rice blast fungus.


Zdroje

1. NikawaJSassPWiglerM 1987 Cloning and characterization of the low-affinity cyclic AMP phosphodiesterase gene of Saccharomyces cerevisiae. Mol Cell Biol 7 3629 3636

2. SassPFieldJNikawaJTodaTWiglerM 1986 Cloning and characterization of the high-affinity cAMP phosphodiesterase of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 83 9303 9307

3. TodaTCameronSSassPZollerMScottJD 1987 Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae. Mol Cell Biol 7 1371 1377

4. TodaTCameronSSassPZollerMWiglerM 1987 Three different genes in Saccharomyces cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase. Cell 50 277 287

5. KimCVigilDAnandGTaylorSS 2006 Structure and dynamics of PKA signaling proteins. Eur J Cell Biol 85 651 654

6. TaylorSSKimCVigilDHasteNMYangJ 2005 Dynamics of signaling by PKA. Biochim Biophys Acta 1754 25 37

7. RispailNSoanesDMAntCCzajkowskiRGrünlerA 2009 Comparative genomics of MAP kinase and calcium-calcineurin signaling components in plant and human pathogenic fungi. Fungal Genet Biol 46 287 298

8. YangZHDickmanMB 1999 Colletotrichum trifolii mutants disrupted in the catalytic subunit of cAMP-dependent protein kinase are nonpathogenic. Mol Plant Microbe Interact 12 430 439

9. TakanoYKomedaKKojimaKOkunoT 2001 Proper regulation of cyclic AMP-dependent protein kinase is required for growth, conidiation, and appressorium function in the anthracnose fungus Colletotrichum lagenarium. Mol Plant Microbe Interact 14 1149 1157

10. YamauchiJTakayanagiNKomedaKTakanoYOkunoT 2004 cAMP-PKA signaling regulates multiple steps of fungal infection cooperatively with Cmk1 MAP kinase in Colletotrichum lagenarium. Mol Plant Microbe Interact 17 1355 1365

11. ChoiYEXuJR 2010 The cAMP signaling pathway in Fusarium verticillioides is important for conidiation, plant infection, and stress responses but not fumonisin production. Mol Plant Microbe Interact 23 522 533

12. 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

13. MitchellTKDeanRA 1995 The cAMP-dependent protein kinase catalytic subunit is required for appressorium formation and pathogenesis by the rice blast pathogen Magnaporthe grisea. Plant Cell 7 1869 1878

14. XuJRUrbanMSweigardJAHamerJE 1997 The CPKA gene of Magnaporthe grisea is essential for appressorial penetration. Mol Plant Microbe Interact 10 187 194

15. JurickWMIIRollinsJA 2007 Deletion of the adenylate cyclase (sac1) gene affects multiple developmental pathways and pathogenicity in Sclerotina sclerotiorum. Fungal Genet Biol 44 521 530

16. GoldSDuncanGBarrettKKronstadJ 1994 cAMP regulates morphogenesis in the fungal pathogen Ustilago maydis. Genes Dev 8 2805 2816

17. DurrenbergerFWongKKronstadJW 1998 Identification of a cAMP-dependent protein kinase catalytic subunit required for virulence and morphogenesis in Ustilago maydis. Proc Natl Acad Sci U S A 95 5684 5689

18. LiuHStylesCAFinkGR 1996 Saccharomyces cerevisiae S288C has a mutation in FLO8, a gene required for filamentous growth. Genetics 144 967 978

19. RuppSSummersELoHJMadhaniHFinkG 1999 MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene. EMBO J 18 1257 1269

20. LoWSDranginisAM 1998 The cell surface flocculin Flo11 is required for pseudohyphae formation and invasion by Saccharomyces cerevisiae. Mol Biol Cell 9 161 171

21. KobayashiOSudaHOhtaniTSoneH 1996 Molecular cloning and analysis of the dominant flocculation gene FLO8 from Saccharomyces cerevisiae. Mol Gen Genet 251 707 715

22. FichtnerLSchulzeFBrausGH 2007 Differential Flo8p-dependent regulation of FLO1 and FLO11 for cell-cell and cell-substrate adherence of Saccharomyces cerevisiae S288c. Mol Microbiol 66 1276 1289

23. PanXHeitmanJ 2002 Protein kinase A operates a molecular switch that governs yeast pseudohyphal differentiation. Mol Cell Biol 22 3981 3993

24. BockmuhlDPErnstJF 2001 A potential phosphorylation site for an A-type kinase in the Efg1 regulator protein contributes to hyphal morphogenesis of Candida albicans. Genetics 157 1523 1530

25. ShenharGKassirY 2001 A positive regulator of mitosis, Sok2, functions as a negative regulator of meiosis in Saccharomyces cerevisiae. Mol Cell Biol 21 1603 1612

26. WardMPGimenoCJFinkGRGarrettS 1995 SOK2 may regulate cyclic AMP-dependent protein kinase-stimulated growth and pseudohyphal development by repressing transcription. Mol Cell Biol 15 6854 6863

27. BiswasSVan-DijckPDattaA 2007 Environmental sensing and signal transduction pathways regulating morphopathogenic determinants of Candida albicans. Microbiol Mol Biol Rev 71 348 376

28. CaoFLaneSRanigaPPLuYZhouZ 2006 The Flo8 transcription factor is essential for hyphal development and virulence in Candida albicans. Mol Biol Cell 17 295 307

29. OharaTTsugeT 2004 FoSTUA, encoding a basic helix-loop-helix protein, differentially regulates development of three kinds of asexual spores, macroconidia, microconidia, and chlamydospores, in the fungal plant pathogen Fusarium oxysporum. Eukaryot Cell 3 1412 1422

30. TongXZZhangXWPlummerKMStowellKMSullivanPA 2007 GcStuA, an APSES transcription factor, is required for generation of appressorial turgor pressure and full pathogenicity of Glomerella cingulata. Mol Plant Microbe Interact 20 1102 1111

31. NishimuraMFukadaJMoriwakiAFujikawaTOhashiM 2009 Mstu1, an APSES transcription factor, is required for appressorium-mediated infection in Magnaporthe grisea. Biosci Biotechnol Biochem 73 1779 1786

32. HartmannHAKahmannRBolkerM 1996 The pheromone response factor coordinates filamentous growth and pathogenicity in Ustilago maydis. EMBO J 15 1632 1641

33. DurrenbergerFLaidlawRDKronstadJW 2001 The hgl1 gene is required for dimorphism and teliospore formation in the fungal pathogen Ustilago maydis. Mol Microbiol 41 337 348

34. LoubradouGBrachmannAFeldbruggeMKahmannR 2001 A homologue of the transcriptional repressor Ssn6p antagonizes cAMP signalling in Ustilago maydis. Mol Microbiol 40 719 730

35. OuSH 1985 Rice diseases. 2nd edition Surrey, UK Commonw. Mycol. Inst

36. HowardRJValentB 1996 Breaking and entering host penetration by the fungal rice blast pathogen Magnaporthe grisea. Annu Rev Microbiol 50 491 512

37. TalbotNJ 2003 On the trail of a cereal killer: exploring the biology of Magnaporthe grisea. Annu Rev Microbiol 57 177 202

38. EbboleDJ 2007 Magnaporthe as a model for understanding host-pathogen interactions. Annu Rev Phytopathol 45 437 56

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

40. XuJRHamerJE 1996 MAP kinase and cAMP signaling regulate infection structure formation and pathogenic growth in the rice blast fungus Magnaporthe grisea. Genes Dev 10 2696 2706

41. XuJRStaigerCJHamerJE 1998 Inactivation of the mitogen-activated protein kinase Mps1 from the rice blast fungus prevents penetration of host cells but allows activation of plant defense responses. Proc Natl Acad Sci U S A 95 12713 12718

42. Caracuel-RiosZTalbotNJ 2007 Cellular differentiation and host invasion by the rice blast fungus Magnaporthe grisea. Curr Opin Microbiol 10 339 345

43. LeeYHDeanRA 1993 cAMP regulates infection structure formation in the plant pathogenic fungus Magnaporthe grisea. Plant Cell 5 693 700

44. AdachiKHamerJE 1998 Divergent cAMP signaling pathways regulate growth and pathogenesis in the rice blast fungus Magnaporthe grisea. Plant Cell 10 1361 1373

45. RamanujamRNaqviNI 2010 PdeH, a high-affinity cAMP phosphodiesterase, is a key regulator of asexual and pathogenic differentiation in Magnaporthe oryzae. PLoS Pathog 6 e1000897

46. ZhaoXKimYParkGXuJR 2005 A mitogen-activated protein kinase cascade regulating infection-related morphogenesis in Magnaporthe grisea. Plant Cell 17 1317 1329

47. ZhaoXXuJR 2007 A highly conserved MAPK-docking site in Mst7 is essential for Pmk1 activation in Magnaporthe grisea. Mol Microbiol 63 881 894

48. ParkGXueCZhaoXKimYOrbachM 2006 Multiple upstream signals converge on the adaptor protein Mst50 in Magnaporthe grisea. Plant Cell 18 2822 2835

49. LiuWZhouXLiGLiLKongL 2011 Multiple plant surface signals are sensed by different mechanisms in the rice blast fungus for appressorium formation. PLoS Pathog 7 e1001261

50. LiuSDeanRA 1997 G protein alpha subunit genes control growth, development, and pathogenicity of Magnaporthe grisea. Mol Plant Microbe Interact 10 1075 1086

51. FangEGDeanRA 2000 Site-directed mutagenesis of the magB gene affects growth and development in Magnaporthe grisea. Mol Plant Microbe Interact 13 1214 1227

52. LiuHSureshAWillardFSSiderovskiDPLuS 2007 Rgs1 regulates multiple Galpha subunits in Magnaporthe pathogenesis, asexual growth and thigmotropism. EMBO J 26 690 700

53. NishimuraMParkGXuJR 2003 The G-beta subunit Mgb1 is involved in regulating multiple steps of infection-related morphogenesis in Magnaporthe grisea. Mol Microbiol 50 231 243

54. LiangSWangZYLiuPJLiDB 2006 A Gγ subunit promoter T-DNA insertion mutant-A1-412 of Magnaporthe grisea is defective in appressorium formation, penetration and pathogenicity. Chinese Sc Bull 51 2214 2218

55. DeZwaanTMCarrollAMValentBSweigardJA 1999 Magnaporthe grisea pth11p is a novel plasma membrane protein that mediates appressorium differentiation in response to inductive substrate cues. Plant Cell 11 2013 2030

56. LiYYanXWangHLiangSMaWB 2010 MoRic8 is a novel component of G-protein signaling during plant infection by the rice blast fungus Magnaporthe oryzae. Mol Plant Microbe Interact 23 217 331

57. CullenPJSabbaghWJrGrahamEIrickMMvan OldenEK 2004 A signaling mucin at the head of the Cdc42- and MAPK-dependent filamentous growth pathway in yeast. Genes Dev 18 1695 1708

58. TalbotNJEbboleDJHamerJE 1993 Identification and characterization of MPG1, a gene involved in pathogenicity from the rice blast fungus Magnaporthe grisea. Plant Cell 5 1575 1590

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

60. LiYLiangSYanXWangHLiDB 2010 Characterization of MoLDB1 required for vegetative growth, infection-related morphogenesis, and pathogenicity in the rice blast fungus Magnaporthe oryzae. Mol Plant Microbe Interact 23 1260 1274

61. OdenbachDBrethBThinesEWeberRWAnkeH 2007 The transcription factor Con7p is a central regulator of infection-related morphogenesis in the rice blast fungus Magnaporthe grisea. Mol Microbiol 64 293 307

62. KimSParkSYKimKSRhoHSChiMH 2009 Homeobox transcription factors are required for conidiation and appressorium development in the rice blast fungus Magnaporthe oryzae. PLoS Genet 5 e1000757

63. LiuWXieSZhaoXChenXZhengW 2010 A homeobox gene is essential for conidiogenesis of the rice blast fungus Magnaporthe oryzae. Mol Plant Microbe Interact 23 366 375

64. LeeBYHanSYChoiHGKimJHHanKH 2005 Screening of growth- or development-related genes by using genomic library with inducible promoter in Aspergillus nidulans. J Microbiol 43 523 528

65. LiGZhouXKongLWangYZhangH 2011 MoSfl1 is important for virulence and heat tolerance in Magnaporthe oryzae. PLoS One 6 e19951

66. StoldtVRSonnebornALeukerCEErnstJF 1997 Efg1p, an essential regulator of morphogenesis of the human pathogen Candida albicans, is a member of a conserved class of bHLH proteins regulating morphogenetic processes in fungi. EMBO J 16 1982 1991

67. EmesRDPontingCP 2001 A new sequence motif linking lissencephaly, Treacher Collins and oral-facial-digital type 1 syndromes, microtubule dynamics and cell migration. Hum Mol Gen 10 2813 2820

68. ParkGXueCZhengLLamSXuJR 2002 MST12 regulates infectious growth but not appressorium formation in the rice blast fungus Magnaporthe grisea. Mol Plant Microbe Interact 15 183 192

69. MehrabiRDingSXuJR 2008 MADS-box transcription factor mig1 is required for infectious growth in Magnaporthe grisea. Eukaryot Cell 7 791 799

70. YangJZhaoXSunJKangZDingS 2010 A novel protein Com1 is required for normal conidium morphology and full virulence in Magnaporthe oryzae. Mol Plant Microbe Interact 23 112 123

71. GuoMChenYDuYDongYGuoW 2011 The bZIP transcription factor MoAP1 mediates the oxidative stress response and is critical for pathogenicity of the rice blast Fungus Magnaporthe oryzae. PLoS Pathog 7 e1001302

72. ZhouXLiuWWangCXuQWangY 2011 A MADS-box transcription factor MoMcm1 is required for male fertility, microconidium production and virulence in Magnaporthe oryzae. Mol Microbiol 80 33 53

73. TuckerSLBesiMIGalhanoRFranceschettiMGoetzS 2010 Common genetic pathways regulate organ-specific infection-related development in the rice blast fungus. Plant Cell 22 953 972

74. LeungHBorromeoESBernardoMANotteghemJL 1988 Genetic analysis of virulence in the rice blast fungus Magnaporthe grisea. Phytopathology 78 1227 1233

75. SambrookJFritschEFManiatisT 1989 Molecular Cloning: A Laboratory Manual Cold Spring Harbor, NY Cold Spring Harbor Laboratory Press

76. CarrollAMSweigardJAValentB 1994 Improved vectors for selecting resistance to hygromycin. Fungal Genet Newsl 41 22

77. Maggio-HallLAKellerNP 2004 Mitochondrial β-oxidation in Aspergillus nidulans. Mol Microbiol 54 1173 1185

78. SweigardJACarrollAMFarrallLValentB 1997 A series of vectors for fungal transformation. Fungal Genet Newsl 44 52 53

79. WangZYSoanesDMKershawMJTalbotNJ 2007 Functional analysis of lipid metabolism in Magnaporthe grisea reveals a role for peroxisomal fatty acid β-oxidation during appressorium-mediated plant infection. Mol Plant Microbe Interact 20 475 491

80. LivakKJSchmittgenTD 2001 Analysis of relative gene expression data using real-time quantitive PCR and the 2−ΔΔCT method. Methods 25 402 408

81. LiuYGChenY 2007 High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences. Biotechniques 43 649 650

82. ChenJZhengWZhengSZhangDSangW 2008 Rac1 is required for pathogenicity and Chm1-dependent conidiogenesis in rice fungal pathogen Magnaporthe grisea. PLoS Pathog 4 e1000202

83. ChumleyFGValentB 1990 Genetic analysis of melanin deficient, nonpathogenic mutants of Magnaporthe grisea. Mol Plant Microbe Interact 3 135 143

84. FosterAJJenkinsonJMTalbotNJ 2003 Trehalose synthesis and metabolism are required at different stages of plant infection by Magnaporthe grisea. EMBO J 22 225 235

85. JeongJSMitchellTKDeanRA 2007 The Magnaporthe grisea snodprot1 homolog, MSP1, is required for virulence. FEMS Microbiol Lett 273 157 165

86. ZhouZLiGLinCHeC 2009 Conidiophore stalk-less1 encodes a putative zinc-finger protein involved in the early stage of conidiation and mycelial infection in Magnaporthe oryzae. Mol Plant Microbe Interact 22 402 410

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 12
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#