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Microwounding is a pivotal factor for the induction of actin-dependent penetration resistance against fungal attack

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Abstract

Induced penetration resistance is triggered by failed penetration attempts of nonpathogenic fungi. The resistance mechanism is an important nonhost reaction in plants that can block the invasion of filamentous pathogens such as fungi and oomycetes. However, it remains unclear whether the mechanical stimuli accompanying fungal penetration play a role in induced penetration resistance, whereas the perforation of the cell wall may provide significant stimuli to plant cells. Here, we used microneedles or biolistic bombardment to mimic fungal penetration pegs and a micromanipulation transfer technique of the bio-probe, a germling of Blumeria graminis hordei, to the wounded cells to demonstrate that microwounds derived from fungal penetration attempts may trigger induced penetration resistance in plant cells. When preinoculated with the nonpathogenic fungi Erysiphe pisi and Colletotrichum orbiculare, which were unable to penetrate a barley cell, the penetration of a bio-probe that was transferred by micromanipulation onto the same cell was completely blocked. Fungal penetration was essential to the triggering of induced penetration resistance because a penetration-peg-defective mutant of C. orbiculare completely lacked the ability to trigger resistance. The artificial microwounds significantly, but not completely, blocked the penetration of the bio-probe. Treatment with the actin polymerization inhibitor cytochalasin A or expression of the actin depolymerizing protein HvPro1 caused complete ablation of the induced penetration resistance triggered by either failed fungal penetration or artificial microwounds. These results strongly suggest that microwounding may trigger actin-dependent induced penetration resistance. Manipulation of induced penetration resistance may be a promising target to improve basic disease resistance in plants.

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Abbreviations

DAMP:

Damage-associated molecular pattern

DIC:

Differential interference contrast

Loc :

Mutant with loss of cellulase secretion

Lop :

Mutant with loss of penetration peg formation

MAMP:

Microbe-associated molecular pattern

PAMP:

Pathogen-associated molecular pattern

References

  • Aist JR (1976) Papillae and related wound plugs of plant cells. Annu Rev Phytopathol 14:145–163

    Article  Google Scholar 

  • Chen Z, Nunes MA, Silva MC, Rodrigues CJ Jr (2004) Appressorium turgor pressure of Colletotrichum kahawae might have a role in coffee cuticle penetration. Mycologia 96:1199–1208

    Article  PubMed  Google Scholar 

  • Clement JA, Martin SG, Porter R, Butt TM, Beckett A (1994) The role of hydrophobicity in attachment of urediniospores and sporelings of Uromyces. Germination and the role of extracellular matrix in adhesion of urediniospores of Uromyces viciae-fabae. Mycol Res 98:1217–1228

    Article  Google Scholar 

  • Conrath U (2011) Molecular aspects of defence priming. Trends Plant Sci 16:524–531

    Article  PubMed  CAS  Google Scholar 

  • Collins NC, Thordal-Christensen H, Lipka V, Bau S, Kombrink E, Qiu JL, Hückelhoven R, Stein M, Freialdenhoven A, Somerville SC, Schulze-Lefert P (2003) SNARE-protein-mediated disease resistance at the plant cell wall. Nature 425:973–977

    Google Scholar 

  • Davis KR, Hahlbrock K (1987) Induction of defense responses in cultured parsley cells by plant cell wall fragments. Plant Physiol 84:1286–1290

    Article  PubMed  CAS  Google Scholar 

  • Enkerli J, Felix G, Boller T (1999) The enzymatic activity of fungal xylanase is not necessary for its elicitor activity. Plant Physiol 121:391–397

    Article  PubMed  CAS  Google Scholar 

  • Fauth M, Schweizer P, Buchala A, Markstädter C, Riederer M, Kato T, Kauss H (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

    Article  PubMed  CAS  Google Scholar 

  • Feng J, Wang F, Liu G, Greenshields D, Shen W, Kaminskyj S, Hughes GR, Peng Y, Selvaraj G, Zou J, Wei Y (2009) Analysis of a Blumeria graminis-secreted lipase reveals the importance of host epicuticular wax components for fungal adhesion and development. Mol Plant Microbe Interact 22:1601–1610

    Article  PubMed  CAS  Google Scholar 

  • Galletti R, Denoux C, Gambetta S, Dewdney J, Ausubel FM, De Lorenzo G, Ferrari S (2008) The AtrbohD-mediated oxidative burst elicited by oligogalacturonides in Arabidopsis is dispensable for the activation of defense responses effective against Botrytis cinerea. Plant Physiol 148:1695–1706

    Article  PubMed  CAS  Google Scholar 

  • Gus-Mayer S, Naton B, Hahlbrock K, Schmelzer E (1998) Local mechanical stimulation induces components of the pathogen defense response in parsley. Proc Natl Acad Sci USA 95:8398–8403

    Article  PubMed  CAS  Google Scholar 

  • Hardham AR, Jones DA, Takemoto D (2007) Cytoskeleton and cell wall function in penetration resistance. Curr Opin Plant Biol 10:342–348

    Article  PubMed  CAS  Google Scholar 

  • Haswell ES, Peyronnet R, Barbier-Brygoo H, Meyerowitz EM, Frachisse JM (2008) Two MscS homologs provide mechanosensitive channel activities in the Arabidopsis root. Curr Biol 18:730–734

    Article  PubMed  CAS  Google Scholar 

  • Howard RJ, Ferrari MA, Roach DH, Money NP (1991) Penetration of hard substrates by a fungus employing enormous turgor pressures. Proc Natl Acad Sci USA 88:11281–11284

    Article  PubMed  CAS  Google Scholar 

  • Jung HW, Tschaplinski TJ, Wang L, Glazebrook J, Greenberg JT (2009) Priming in systemic plant immunity. Science 324:89–91

    Article  PubMed  Google Scholar 

  • Katoh M, Hirose I, Kubo Y, Hikichi Y, Kunoh H, Furusawa I, Shishiyama J (1988) Use of mutants to indicate factors prerequisite for penetration of Colletotrichum lagenarium by appressoria. Physiol Mol Plant Pathol 32:177–184

    Article  Google Scholar 

  • Kobayashi I, Komura T, Sakamoto Y, Yamaoka N, Kunoh H (1990) Recognition of a pathogen and a nonpathogen by barley coleoptile cells. (I) Cytoplasmic responses to the nonpathogen, Erysiphe pisi, prior to its penetration. Physiol Mol Plant Pathol 37:479–490

    Article  Google Scholar 

  • Kobayashi I, Watanabe H, Kunoh H (1995) Induced accessibility and enhanced inaccessibility at the cellular level in barley coleoptiles. XIV Evidence for elicitor(s) and suppressor(s) of host inaccessibility from Erysiphe graminis. Physiol Mol Plant Pathol 46:445–456

    Article  Google Scholar 

  • Kobayashi Y, Kobayashi I, Funaki Y, Fujimoto S, Takemoto T, Kunoh H (1997) Dynamic reorganization of microfilaments and microtubules is necessary for the expression of non-host resistance in barley coleoptile cells. Plant J 11:525–537

    Article  CAS  Google Scholar 

  • Kobayashi I, Yamada M, Kobayashi Y (2007) Calcium ion promotes successful penetration of powdery mildew fungi into barley cells. J Gen Plant Pathol 73:399–404

    Article  CAS  Google Scholar 

  • Kováts K, Binder A, Hohl HR (1991) Cytology of induced systemic resistance of cucumber to Colletotrichum lagenarium. Planta 183:484–490

    Google Scholar 

  • Kunoh H, Hayashimoto A, Harui M, Ishizaki H (1985) Induced accessibility and enhanced resistance at the cellular level in barley coleoptiles. I The significance of timing of fungal invasion. Physiol Plant Pathol 27:43–54

    Article  Google Scholar 

  • Kunoh H, Katsuragawa N, Yamaoka N, Hayashimoto A (1988) Induced accessibility and enhanced inaccessibility at the cellular level in barley coleoptiles. III Timing and localization of enhanced inaccessibility in a single coleoptile cell and its transfer to an adjacent cell. Physiol Mol Plant Pathol 33:81–93

    Article  Google Scholar 

  • Kunoh H, Toyoda K, Yamaoka N, Kobayashi I (1989) Induced accessibility and enhanced inaccessibility at the cellular level in barley coleoptiles. V Duration of stimulus by a non-pathogen in relation to enhanced inaccessibility. Physiol Mol Plant Pathol 35:507–518

    Article  Google Scholar 

  • Lipka V, Dittgen J, Bednarek P, Bhat R, Wiermer M, Stein M, Landtag J, Brandt W, Rosahl S, Scheel D, Llorente F, Molina A, Parker J, Somerville S, Schulze-Lefert P (2005) Pre- and postinvasion defenses both contribute to nonhost resistance in Arabidopsis. Science 310:1180–1183

    Google Scholar 

  • Lu JP, Liu XH, Feng XX, Min H, Lin FC (2009) An autophagy gene, MgATG5, is required for cell differentiation and pathogenesis in Magnaporthe oryzae. Curr Genet 55:461–473

    Article  PubMed  CAS  Google Scholar 

  • Matern U, Grimmig B, Kneusel RE (1995) Plant cell wall reinforcement in the disease resistance response—molecular composition and regulation. Can J Bot 73:S511–S517

    Article  CAS  Google Scholar 

  • Nakagawa Y, Katagiri T, Shinozaki K, Qi Z, Tatsumi H, Furuichi T, Kishigami A, Sokabe M, Kojima I, Sato S, Kato T, Tabata S, Iida K, Terashima A, Nakano M, Ikeda M, Yamanaka T, Iida H (2007) Arabidopsis plasma membrane protein crucial for Ca2+ influx and touch sensing in roots. Proc Natl Acad Sci USA 104:3639–3644

    Article  PubMed  CAS  Google Scholar 

  • Nguyen QB, Itoh K, Van Vu B, Tosa Y, Nakayashiki H (2011) Simultaneous silencing of endo-β-1,4 xylanase genes reveals their roles in the virulence of Magnaporthe oryzae. Mol Microbiol 81:1008–1019

    Article  PubMed  CAS  Google Scholar 

  • Nielsen KA, Nicholson RL, Carver TLW, Kunoh H, Oliver RP (2000) First touch: an immediate response to surface recognition in conidia of Blumeria graminis. Physiol Mol Plant Pathol 56:63–70

    Article  Google Scholar 

  • Niwa Y, Hirano T, Yoshimoto K, Shimizu M, Kobayashi H (1999) Non-invasive quantitative detection and applications of non-toxic S65T-type green fluorescent protein in living plants. Plant J 18:455–463

    Article  PubMed  CAS  Google Scholar 

  • Park G, Bruno KS, Staiger CJ, Talbot NJ, Xu JR (2004) Independent genetic mechanisms mediate turgor generation and penetration peg formation during plant infection in the rice blast fungus. Mol Microbiol 53:1695–1707

    Article  PubMed  CAS  Google Scholar 

  • Perpetua NS, Kubo Y, Okuno T, Furusawa I (1994) Restoration of pathogenicity of a penetration-deficient mutant of Collectotrichum lagenarium by DNA complementation. Curr Genet 25:41–46

    Article  PubMed  CAS  Google Scholar 

  • Prime-A-Plant Group, Conrath U, Beckers GJ, Flors V, García-Agustín P, Jakab G, Mauch F, Newman MA, Pieterse CM, Poinssot B, Pozo MJ, Pugin A, Schaffrath U, Ton J, Wendehenne D, Zimmerli L, Mauch-Mani B (2006) Priming: getting ready for battle. Mol Plant Microbe Interact 19:1062–1071

    Article  PubMed  CAS  Google Scholar 

  • Schulze-Lefert P (2004) Knocking on the heaven’s wall: pathogenesis of and resistance to biotrophic fungi at the cell wall. Curr Opin Plant Biol 7:377–383

    Article  PubMed  CAS  Google Scholar 

  • Shimada C, Lipka V, O’Connell R, Okuno T, Schulze-Lefert P, Takano Y (2006) Nonhost resistance in Arabidopsis–Colletotrichum interactions acts at the cell periphery and requires actin filament function. Mol Plant Microbe Interact 19:270–279

    Article  PubMed  CAS  Google Scholar 

  • Shimono M, Sugano S, Nakayama A, Jiang CJ, Ono K, Toki S, Takatsuji H (2007) Rice WRKY45 plays a crucial role in benzothiadiazole-inducible blast resistance. Plant Cell 19:2064–2076

    Article  PubMed  CAS  Google Scholar 

  • Shimono M, Koga H, Akagi A, Hayashi N, Goto S, Sawada M, Kurihara T, Matsushita A, Sugano S, Jiang CJ, Kaku H, Inoue H, Takatsuji H (2012) Rice WRKY45 plays important roles in fungal and bacterial disease resistance. Mol Plant Pathol 13:83–94

    Article  PubMed  CAS  Google Scholar 

  • Skamnioti P, Gurr SJ (2007) Magnaporthe grisea cutinase2 mediates appressorium differentiation and host penetration and is required for full virulence. Plant Cell 19:2674–2689

    Article  PubMed  CAS  Google Scholar 

  • Staiger CJ, Yuan M, Valenta R, Shaw PJ, Warn RM, Lloyd CW (1994) Microinjected profilin affects cytoplasmic streaming in plant cells by rapidly depolymerizing actin microfilaments. Curr Biol 4:215–219

    Article  PubMed  CAS  Google Scholar 

  • Stein M, Dittgen J, Sánchez-Rodríguez C, Hou BH, Molina A, Schulze-Lefert P, Lipka V, Somerville S (2006) Arabidopsis PEN3/PDR8, an ATP binding cassette transporter, contributes to nonhost resistance to inappropriate pathogens that enter by direct penetration. Plant Cell 18:731–746

    Google Scholar 

  • Stumm D, Gessler C (1986) Role of papillae in the induced systemic resistance of cucumber against Colletotrichum lagenarium. Physiol Mol Plant Pathol 29:405–410

    Article  Google Scholar 

  • Takemoto D, Jones DA, Hardham AR (2003) GFP-tagging of cell components reveals the dynamics of subcellular re-organization in response to infection of Arabidopsis by oomycete pathogens. Plant J 33:775–792

    Article  PubMed  CAS  Google Scholar 

  • Tominaga M, Yokota E, Vidali L, Sonobe S, Hepler PK, Shimmen T (2000) The role of plant villin in the organization of the actin cytoskeleton, cytoplasmic streaming and the architecture of the transvacuolar strand in root hair cells of Hydrocharis. Planta 210:836–843

    Article  PubMed  CAS  Google Scholar 

  • Tonukari NJ, Scott-Craig JS, Walton JD (2000) The Cochliobolus carbonum SNF1 gene is required for cell wall-degrading enzyme expression and virulence on maize. Plant Cell 12:237–248

    PubMed  CAS  Google Scholar 

  • Toyoda K, Kobayashi I, Kunoh H (1993) Elicitor activity of a fungal product assessed at the single-cell level by a novel gel-bead method. Plant Cell Physiol 34:775–780

    CAS  Google Scholar 

  • Tsuji G, Fujii S, Tsuge S, Shiraishi T, Kubo Y (2003) The Colletotrichum lagenarium Ste12-like gene CST1 is essential for appressorium penetration. Mol Plant Microbe Interact 16:315–325

    Article  PubMed  CAS  Google Scholar 

  • Underwood W, Somerville SC (2008) Focal accumulation of defences at sites of fungal pathogen attack. J Exp Bot 59:3501–3508

    Article  PubMed  CAS  Google Scholar 

  • Xu JR, Staiger CJ, Hamer JE (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 USA 95:12713–12718

    Article  PubMed  CAS  Google Scholar 

  • Zhang Z, Henderson C, Perfect E, Carver TLW, Thomas BJ, Skamnioti P, Gurr SJ (2005) Of genes and genomes, needles and haystacks: Blumeria graminis and functionality. Mol Plant Pathol 6:561–575

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Dr. Yasuyuki Kubo (Laboratory of Plant Pathology, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto, Japan) for the gift of Colletotrichum orbiculare mutants. This work was supported in part by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Integrated research project for plant, insect and animal using genome technology PMI0006 and Grant-in-Aid for Scientific Research No. 23580062 (2011–2013) from the Ministry of Education, Science and Culture of Japan, both given to I. Kobayashi.

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Authors and Affiliations

  1. Core-Lab, Graduate School of Regional Innovation Studies, Mie University, 1577 Kurima-Machiya-cho, Tsu, Mie, 514-8507, Japan

    Yuhko Kobayashi & Issei Kobayashi

  2. Division of Plant Functional Genomics, Life Science Research Center, Mie University, 1577 Kurima-Machiya-cho, Tsu, Mie, 514-8507, Japan

    Issei Kobayashi

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  1. Yuhko Kobayashi
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  2. Issei Kobayashi
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Correspondence to Issei Kobayashi.

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Kobayashi, Y., Kobayashi, I. Microwounding is a pivotal factor for the induction of actin-dependent penetration resistance against fungal attack. Planta 237, 1187–1198 (2013). https://doi.org/10.1007/s00425-013-1837-9

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  • DOI: https://doi.org/10.1007/s00425-013-1837-9

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