A Permeable Cuticle Is Associated with the Release of Reactive Oxygen Species and Induction of Innate Immunity


Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS), including H2O2 and O2, are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI) or catalase. H2O2 was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA). Accordingly, ABA biosynthesis mutants (aba2 and aba3) were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending innate defenses.


Vyšlo v časopise: A Permeable Cuticle Is Associated with the Release of Reactive Oxygen Species and Induction of Innate Immunity. PLoS Pathog 7(7): e32767. doi:10.1371/journal.ppat.1002148
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002148

Souhrn

Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS), including H2O2 and O2, are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI) or catalase. H2O2 was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA). Accordingly, ABA biosynthesis mutants (aba2 and aba3) were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending innate defenses.


Zdroje

1. RiedererMSchreiberL 2001 Protecting against water loss: analysis of the barrier properties of plant cuticles. J Exp Bot 52 2023 2032

2. NawrathC 2006 Unraveling the complex network of cuticular structure and function. Curr Opin Pl Biol 9 281 287

3. GoodwinSMJenksMA 2005 Plant cuticle function as a barrier to water loss. Plant cuticle function as a barrier to water loss. JenksMAHasegawaPM Oxford, UK Blackwell Publishing Inc 14 36

4. KunstLSamuelsL 2009 Plant cuticles shine: advances in wax biosynthesis and export. Cur Opin Plant Biol 12 721 727

5. SamuelsLKunstLJetterR 2008 Sealing plant surfaces: Cuticular wax formation by epidermal cells. Annu Rev Plant Biol 59 683 707

6. KolattukudyPERogersLMLiDXHwangCSFlaishmanMA 1995 Surface signaling in pathogenesis. Proc Natl Acad Sci USA 92 4080 4087

7. HerediaA 2003 Biophysical and biochemical characteristics of cutin, a plant barrier biopolymer. Bioch Biophys Acta 1620 1 7

8. PollardMBeissonFLiYOhlroggeJB 2008 Building lipid barriers: biosynthesis of cutin and suberin. Trends Plant Sci 13 236 246

9. CommenilPBelingheriLDehorterB 1998 Antilipase antibodies prevent infection of tomato leaves by Botrytis cinerea. Phys Mol Plant Pathol 52 1 14

10. MaitiIBKolattukudyPE 1979 Prevention of fungal infection of plants by specific-inhibition of cutinase. Science 205 507 508

11. LiDAshbyAMJohnstoneK 2003 Molecular evidence that the extracellular cutinase Pbc1 is required for pathogenicity of Pyrenopeziza brassicae on oilseed rape. Mol Plant Microbe Interact 16: 16 545 552

12. RogersLMFlaishmanMAKolattukudyPE 1994 Cutinase gene disruption in Fusarium solani f sp pisi decreases its virulence on pea. Plant Cell 6 935 945

13. ReisHPfiffiSHahnM 2005 Molecular and functional characterization of a secreted lipase from Botrytis cinerea. Mol Plant Pathol 6 257 267

14. StahlDJTheuerkaufAHeitefussRSchaferW 1994 Cutinase of Nectria haematococca (Fusarium solani f sp pisi) is not required for fungal virulence or organ specificity on pea. Mol Plant Microbe Interact 7 713 725

15. SweigardJAChumleyFGValentB 1992 Disruption of a Magnaporthe grisea cutinase gene. Mol Gen Genetics 232 183 190

16. van KanJALvant KloosterJWWagemakersCAMDeesDCTvan der Vlugt BergmansCJB 1997 Cutinase A of Botrytis cinerea is expressed, but not essential, during penetration of gerbera and tomato. Mol Plant Microbe Interact 10 30 38

17. LinTSKolattukudyPE 1978 Induction of a bio-polyester hydrolase (cutinase) by low-levels of cutin monomers in Fusarium solani f sp pisi. J Bacteriol 133 942 951

18. WoloshukCPKolattukudyPE 1986 Mechanism by which contact with plant cuticle triggers cutinase gene-expression in the spores of Fusarium solani f sp pisi. Proc Natl Acad Sci USA 83 1704 1708

19. FrancisSADeweyFMGurrSJ 1996 The role of cutinase in germling development and infection by Erysiphe graminis f sp hordei. Physiol Mol Plant Pathol 49 201 211

20. GilbertRDJohnsonAMDeanRA 1996 Chemical signals responsible for appressorium formation in the rice blast fungus Magnaporthe grisea. Physiol Mol Plant Pathol 48 335 346

21. PodilaGKRogersLMKolattukudyPE 1993 Chemical signals from avocado surface wax trigger germination and appressorium formation in Colletotrichum gloeosporioides. Plant Physiol 103 267 272

22. SchweizerPJeanguénatAMétrauxJPMösingerE 1994 Plant protection by free cutin monomers in two cereal pathosystems. DanielsMJDownieJAOsbournAE Advances Mol Genet Plant-Microbe Interact Dordrecht Kluwer 371 374

23. SchweizerPFelixGBuchalaAMullerCMétrauxJP 1996 Perception of free cutin monomers by plant cells. Plant J 10 331 341

24. FauthMSchweizerPBuchalaAMarkstadterCRiedererM 1998 Cutin monomers and surface wax constituents elicit H2O2 in conditioned cucumber hypocotyl segments and enhance the activity of other H2O2 elicitors. Plant Physiol 117 1373 1380

25. ApelKHirtH 2004 Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55 373 399

26. TorresMAJonesJDDanglJL 2006 Reactive oxygen species signaling in response to pathogens. Plant Physiol 141 373 378

27. TorresMA 2010 ROS in biotic interactions. Physiol Plant 138 414 429

28. ParkerDMKollerW 1998 Cutinase and other lipolytic esterases protect bean leaves from infection by Rhizoctonia solani. Mol Plant Microbe Interact 11 514 522

29. SieberPSchorderetMRyserUBuchalaAKolattukudyP 2000 Transgenic Arabidopsis plants expressing a fungal cutinase show alterations in the structure and properties of the cuticle and postgenital organ fusions. Plant Cell 12 721 737

30. WellesenKDurstFPinotFBenvenisteINettesheimK 2001 Functional analysis of the LACERATA gene of Arabidopsis provides evidence for different robes of fatty acid omega-hydroxylation in development. Proc Natl Acad Sci USA 98 9704 9699

31. KurdyukovSFaustANawrathCBarSVoisinD 2006 The epidermis-specific extracellular BODYGUARD controls cuticle development and morphogenesis in Arabidopsis. Plant Cell 18 321 339

32. BessireMChassotCJacquatACHumphryMBorelS 2007 A permeable cuticle in Arabidopsis leads to a strong resistance to Botrytis cinerea. EMBO J 26 2158 2168

33. BollerTFelixG 2009 A renaissance of elicitors: perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Annu Rev Plant Biol 60 379 406

34. ChassotCBuchalaASchoonbeekHMétrauxJPLamotteO 2008 Wounding of Arabidopsis leaves causes a powerful but transient protection against Botrytis infection. Plant J 55 555 567

35. LeonJRojoESanchez-SerranoJJ 2001 Wound signalling in plants. J Exp Bot 52 1 9

36. Orozco-CardenasMLNarvaez-VasquezJRyanCA 2001 Hydrogen peroxide acts as a second messenger for the induction of defense genes in tomato plants in response to wounding, systemin, and methyl jasmonate. Plant Cell 13 179 191

37. HempelSLBuettnerGRO'MalleyYQWesselsDAFlahertyDM 1999 Dihydrofluorescein diacetate is superior for detecting intracellular oxidants: Comparison with 2 ',7 '-dichlorodihydrofluorescein diacetate, 5(and 6)-carboxy-2', 7 '-dichlorodihydrofluorescein diacetate, and dihydrorhodamine 123. Free Radical Biol Medicine 27 146 159

38. WarmELatiesGG 1982 Quantification of hydrogen peroxide in plant extracts by the chemiluminescence reaction with luminol. Phytochemistry 21 827 831

39. Lozano-JusteJLeonJ 2010 Enhanced abscisic acid-mediated responses in nia1nia2noa1-2 triple mutant impaired in NIA/NR- and AtNOA1-dependent nitric oxide biosynthesis in Arabidopsis. Plant Physiol 152 891 903

40. VoisinDNawrathCKurdyukovSFrankeRBReina-PintoJJ 2009 Dissection of the complex phenotype in cuticular mutants of Arabidopsis reveals a role of SERRATE as a mediator. Plos Genetics 5 1 19

41. PrinsTWTudzynskiPvon TiedemannATudzynskiBTen HaveA 2000 Infection strategies of Botrytis cinerea and related necrotrophic pathogens. KronstadJW Fungal Pathology Dordrecht Kluwer Academic Publishers 33 64

42. CessnaSGSearsVEDickmanMBLowPS 2000 Oxalic acid, a pathogenicity factor for Sclerotinia sclerotiorum, suppresses the oxidative burst of the host plant. Plant Cell 12 2191 2200

43. WalzAZingen-SellITheisenSKortekampA 2008 Reactive oxygen intermediates and oxalic acid in the pathogenesis of the necrotrophic fungus Sclerotinia sclerotiorum. Europ J Plant Pathol 120 317 330

44. HoffmannAHammesEPliethCDeselCSattelmacherB 2005 Effect of CO2 supply on formation of reactive oxygen species in Arabidopsis thaliana. Protoplasma 227 3 9

45. KesslerABaldwinIT 2002 Plant responses to insect herbivory: The emerging molecular analysis. Annu Rev Plant Biol 53 299 328

46. Van BaarlenPWolteringEJStaatsMvan KanJAL 2007 Histochemical and genetic analysis of host and non-host interactions of Arabidopsis with three Botrytis species: an important role for cell death control. Mol Plant Pathol 8 41 54

47. ChassotCNawrathCMétrauxJP 2007 Cuticular defects lead to full immunity to a major plant pathogen. Plant J 49 972 980

48. WuYSanchezJPLopez-MolinaLHimmelbachAGrillE 2003 The abi1-1 mutation blocks ABA signaling downstream of cADPR action. Plant J 34 307 315

49. HimmelbachAHoffmannTLeubeMHohenerBGrillE 2002 Homeodomain protein ATHB6 is a target of the protein phosphatase ABI1 and regulates hormone responses in Arabidopsis. EMBO J 21 3029 3038

50. AsselberghBCurversKFrancaSCAudenaertKVuylstekeM 2007 Resistance to Botrytis cinerea in sitiens, an abscisic acid-deficient tomato mutant, involves timely production of hydrogen peroxide and cell wall modifications in the epidermis. Plant Physiol 144 1863 1877

51. Mauch-ManiBMauchF 2005 The role of abscisic acid in plant-pathogen interactions. Curr Opin Plant Biol 8 409 414

52. RaghavendraASGonuguntaVKChristmannAGrillE 2010 ABA perception and signalling. Trends Plant Sci 15 395 401

53. ChassotCNawrathCMétrauxJP 2008 The cuticle: not only a barrier for defence. Plant Signal Behav 3 142 144

54. CurversKSeifiHMouilleGDe RyckeRAsselberghB 2010 ABA-deficiency causes changes in cuticle permeability and pectin composition that influence tomato resistance to Botrytis cinerea. Plant Physiol 154 847 860

55. MétrauxJ-PJacksonRWEstherSGoldbachRW 2009 Plant pathogens as suppressors of host defense. Van LoonLC Advances in Botanical Research Academic Press 39 89

56. SchoonbeekHJacquat-BovetACMascherFMétrauxJP 2007 Oxalate-degrading bacteria can protect Arabidopsis thaliana and crop plants against Botrytis cinerea. Mol Plant Microbe Interact 20 1535 1544

57. WalzAZingen-SellILoefflerMSauerM 2008 Expression of an oxalate oxidase gene in tomato and severity of disease caused by Botrytis cinerea and Sclerotinia sclerotiorum. Plant Pathol 57 453 458

58. SiewersVKokkelinkLSmedsgaardJTudzynskiP 2006 Identification of an abscisic acid gene cluster in the grey mold Botrytis cinerea. Appl Environ Microbiol 72 4619 4626

59. SiewersVSmedsgaardJTudzynskiP 2004 The p450 monooxygenase BcABA1 is essential for abscisic acid biosynthesis in Botrytis cinerea. Appl Environ Microbiol 70 3868 3876

60. KettnerJDörfflingK 1995 Biosynthesis and metabolism of abscisic acid in tomato leaves infected with Botrytis cinerea. Planta 196 627 634

61. TemmeNTudzynskiP 2009 Does Botrytis cinerea ignore H2O2-induced oxidative stress during infection? Characterization of BOTRYTIS ACTIVATOR PROTEIN 1. Mol Plant Microbe Interact 22 987 998

62. Léon-KloosterzielKMGilMARuijsGJJacobsenSEOlszewskiNE 1996 Isolation and characterization of abscisic acid-deficient Arabidopsis mutants at two new loci. Plant J 10 655 661

63. MengisteTChenXSalmeronJDietrichR 2003 The BOTRYTIS SUSCEPTIBLE1 gene encodes an R2R3MYB transcription factor protein that is required for biotic and abiotic stress responses in Arabidopsis. Plant Cell 15 2551 2565

64. Thordal-ChristensenHZhangZWeiYCollingeDB 1997 Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley powdery mildew interaction. Plant J 11 1187 1194

65. ChenZXSilvaHKlessigDF 1993 Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science 262 1883 1886

66. ChristmannAHoffmannTTeplovaIGrillEMullerA 2005 Generation of active pools of abscisic acid revealed by in vivo imaging of water-stressed Arabidopsis. Plant Physiol 137 209 219

67. CzechowskiTStittMAltmannTUdvardiMKScheibleWR 2005 Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139 5 17

68. Van GemerenIAMustersWVan Den HondelCAMJJVerripsCT 1995 Construction and heterologous expression of a synthetic copy of the cutinase cDNA from Fusarium solani pisi. J Biotech 40 155 162

79. FrankeRBriesenIWojciechowskiTFaustAYephremovA 2005 Apoplastic polyesters in Arabidopsis surface tissues - A typical suberin and a particular cutin. Phytochemistry 66 2643 2658

70. CloughSJBentAF 1998 Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16 735 743

71. LangELangH 1972 Specific color-reaction for direct identification of formic-acid. Fresenius Zeitschr Anal Chemie 260 8 10

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 7
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Eozinofilní granulomatóza s polyangiitidou
nový kurz
Autori: doc. MUDr. Martina Doubková, Ph.D.

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