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Constitutively activated barley ROPs modulate epidermal cell size, defense reactions and interactions with fungal leaf pathogens

  • Biotic and Abiotic Stress
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Abstract

RHO-like monomeric G-proteins of plants (ROPs, also called RACs), are involved in plant development and interaction with the environment. The barley (Hordeum vulgare) ROP protein HvRACB has been shown to be required for entry of the biotrophic powdery mildew fungus Blumeria graminis f.sp. hordei (Bgh) into living host cells. To get a deeper insight into evolutionarily conserved functions of ROPs in cell polarity and pathogen responses, we stably expressed constitutively activated (CA) mutant variants of different barley ROPs (HvRACB, HvRAC1, HvRAC3) in barley. CA HvROPs induced epidermal cell expansion and/or abolished polarity in tip growing root hairs. All three CA HvROPs enhanced susceptibility of barley to penetration by Bgh whereas only CA HvRAC1 supported whole cell H2O2 production in non-penetrated cells. Despite increasing penetration by Bgh, CA HvRAC1 promoted callose deposition at sites of fungal attack and resistance to penetration by Magnaporthe oryzae. The data show an involvement of ROPs in polar growth processes of the monocot barley and in responses to fungal pathogens with different life style.

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Abbreviations

CA:

Constitutively activated

CWA:

Cell wall apposition

DAB:

3,3-Diaminobenzidine

HAI:

Hours after inoculation

HR:

Hypersensitive reaction

MLO:

MILDEW LOCUS O

ROS:

Reactive oxygen species

RHO:

Rat sarcome oncogene product (RAS) homologue

ROP:

RHO of plants

RAC:

Ras related C3 botulinum toxin substrate

References

  • Agrawal GK, Iwahashi H, Rakwal R (2003) Small GTPase ‘Rop’: molecular switch for plant defense responses. FEBS Lett 546:173–180

    Article  PubMed  CAS  Google Scholar 

  • Berken A (2006) ROPs in the spotlight of plant signal transduction. Cell Mol Life Sci 63:2446–2459

    Article  PubMed  CAS  Google Scholar 

  • Bloch D, Lavy M, Efrat Y, Efroni I, Bracha-Drori K, Abu-Abied M, Sadot E, Yalovsky S (2005) Ectopic expression of an activated RAC in Arabidopsis disrupts membrane cycling. Mol Biol Cell 16:1913–1927

    Article  PubMed  CAS  Google Scholar 

  • Carol RJ, Takeda S, Linstead P, Durrant MC, Kakesova H, Derbyshire P, Drea S, Zarsky V, Dolan L (2005) A RhoGDP dissociation inhibitor spatially regulates growth in root hair cells. Nature 438:1013–1016

    Article  PubMed  CAS  Google Scholar 

  • Christensen TM, Vejlupkova Z, Sharma YK, Arthur KM, Spatafora JW, Albright CA, Meeley RB, Duvick JP, Quatrano RS, Fowler JE (2003) Conserved subgroups and developmental regulation in the monocot rop gene family. Plant Physiol 133:1791–1808

    Article  PubMed  CAS  Google Scholar 

  • Couch BC, Kohn LM (2002) A multilocus gene genealogy concordant with host preference indicates segregation of a new species, Magnaporthe oryzae, from M. grisea. Mycologia 94:683–693

    Article  CAS  Google Scholar 

  • Foreman J, Demidchik V, Bothwell JH, Mylona P, Miedema H, Torres MA, Linstead P, Costa S, Brownlee C, Jones JD, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446

    Article  PubMed  CAS  Google Scholar 

  • Fu Y, Li H, Yang Z (2002) The ROP2 GTPase controls the formation of cortical fine F-actin and the early phase of directional cell expansion during Arabidopsis organogenesis. Plant Cell 14:777–794

    Article  PubMed  CAS  Google Scholar 

  • Fu Y, Gu Y, Zheng Z, Wasteneys G, Yang Z (2005) Arabidopsis interdigitating cell growth requires two antagonistic pathways with opposing action on cell morphogenesis. Cell 120:687–700

    Article  PubMed  CAS  Google Scholar 

  • Fujiwara M, Umemura K, Kawasaki T, Shimamoto K (2006) Proteomics of Rac GTPase signaling reveals its predominant role in elicitor-induced defense response of cultured rice cells. Plant Physiol 140:734–745

    Article  PubMed  CAS  Google Scholar 

  • Görg R, Hollricher K, Schulze-Lefert P (1993) Functional analysis and RFLP-mediated mapping of the Mlg resistance locus in barley. Plant J 3:857–866

    Article  Google Scholar 

  • Gruenheid S, Finlay BB (2003) Microbial pathogenesis and cytoskeletal function. Nature 422:775–781

    Article  PubMed  CAS  Google Scholar 

  • Hassanain HH, Sharma YK, Moldovan L, Khramtsov V, Berliner LJ, Duvick JP, Goldschmidt-Clermont PJ (2000) Plant Rac proteins induce superoxide production in mammalian cells. Biochem Biophys Res Commun 272:783–788

    Article  PubMed  CAS  Google Scholar 

  • Hensel G, Valkov V, Middlefell-Williams J, Kumlehn J (2008) Efficient generation of transgenic barley: the way forward to modulate plant-microbe interactions. J Plant Physiol 165:71–82

    Article  PubMed  CAS  Google Scholar 

  • Hong Z, Zhang Z, Olson JM, Verma DP (2001) A novel UDP-glucose transferase is part of the callose synthase complex and interacts with phragmoplastin at the forming cell plate. Plant Cell 13:769–779

    Article  PubMed  CAS  Google Scholar 

  • Hückelhoven R (2007) Cell wall-associated mechanisms of disease resistance and susceptibility. Annu Rev Phytopathol 45:101–127

    Article  PubMed  CAS  Google Scholar 

  • Hückelhoven R, Kogel KH (1998) Tissue-specific superoxide generation at interaction sites in resistant and susceptible near-isogenic barley lines attacked by the powdery mildew fungus (Erysiphe graminis f.sp. hordei). Mol Plant Microbe Interact 11:292–300

    Article  Google Scholar 

  • Hückelhoven R, Fodor J, Preis C, Kogel KH (1999) Hypersensitive cell death and papilla formation in barley attacked by the powdery mildew fungus are associated with H2O2 but not with salicylic acid accumulation. Plant Physiol 119:1251–1260

    Article  PubMed  Google Scholar 

  • Hwang JU, Gu Y, Lee YJ, Yang Z (2005) Oscillatory ROP GTPase activation leads the oscillatory polarized growth of pollen tubes. Mol Biol Cell 16:5385–5399

    Article  PubMed  CAS  Google Scholar 

  • Jacobs AK, Lipka V, Burton RA, Panstruga R, Strizhov N, Schulze-Lefert P, Fincher GB (2003) An Arabidopsis callose synthase, GSL5, is required for wound and papillary callose formation. Plant Cell 15:2503–2513

    Article  PubMed  CAS  Google Scholar 

  • Jarosch B, Kogel KH, Schaffrath U (1999) The ambivalence of the barley Mlo Locus: mutations conferring resistance against powdery mildew (Blumeria graminis f.sp. hordei) enhance susceptibility to the rice blast fungus Magnaporthe grisea. Mol Plant Microbe Interact 12:508–514

    Article  CAS  Google Scholar 

  • Jones MA, Shen JJ, Fu Y, Li H, Yang Z, Grierson CS (2002) The Arabidopsis Rop2 GTPase is a positive regulator of both root hair initiation and tip growth. Plant Cell 14:763–776

    Article  PubMed  CAS  Google Scholar 

  • Jones MA, Raymond MJ, Yang Z, Smirnoff N (2007) NADPH oxidase-dependent reactive oxygen species formation required for root hair growth depends on ROP GTPase. J Exp Bot 58:1261–1270

    Article  PubMed  CAS  Google Scholar 

  • Kawasaki T, Koita H, Nakatsubo T, Hasegawa K, Wakabayashi K, Takahashi H, Umemura K, Umezawa T, Shimamoto K (2006) Cinnamoyl-CoA reductase, a key enzyme in lignin biosynthesis, is an effector of small GTPase Rac in defense signaling in rice. Proc Natl Acad Sci USA 103:230–235

    Article  PubMed  CAS  Google Scholar 

  • Kawasaki T, Henmi K, Ono E, Hatakeyama S, Iwano M, Satoh H, Shimamoto K (1999) The small GTP-binding protein Rac is a regulator of cell death in plants. Proc Natl Acad Sci USA 96:10922–10926

    Article  PubMed  CAS  Google Scholar 

  • Klahre U, Kost B (2006) Tobacco RhoGTPase ACTIVATING PROTEIN1 spatially restricts signaling of RAC/Rop to the apex of pollen tubes. Plant Cell 18:3033–3046

    Article  PubMed  CAS  Google Scholar 

  • Koga H, Bushnell WR, Zeyen RJ (1990) Specificity of cell type and timing of events associated with papilla formation and the hypersensitive reaction in leaves of Hordeum vulgare attacked by Erysiphe graminis f.sp. hordei. Can J Bot 68:2344–2352

    Google Scholar 

  • Liu G, Kennedy R, Greenshields DL, Peng G, Forseille L, Selvaraj G, Wei Y (2007) Detached and attached Arabidopsis leaf assays reveal distinctive defense responses against hemibiotrophic Colletotrichum spp. Mol Plant Microbe Interact 20:1308–1319

    Article  PubMed  CAS  Google Scholar 

  • Lyngkjær MF, Carver TLW (1999) Modification of mlo5 resistance to Blumeria graminis attack in barley as a consequence of induced accessibility and inaccessibility. Physiol Mol Plant Pathol 55:163–174

    Article  Google Scholar 

  • Miklis M, Consonni C, Bhat RA, Lipka V, Schulze-Lefert P, Panstruga R (2007) Barley Mlo modulates actin-dependent and actin-independent antifungal defence pathways at the cell periphery. Plant Physiol 144:1132–1143

    Article  PubMed  CAS  Google Scholar 

  • Moeder W, Yoshioka K, Klessig DF (2005) Involvement of the small GTPase Rac in the defense responses of tobacco to pathogens. Mol Plant Microbe Interact 18:116–124

    Article  PubMed  CAS  Google Scholar 

  • Molendijk AJ, Bischoff F, Rajendrakumar CS, Friml J, Braun M, Gilroy S, Palme K (2001) Arabidopsis thaliana Rop GTPases are localized to tips of root hairs and control polar growth. EMBO J 20:2779–2788

    Article  PubMed  CAS  Google Scholar 

  • Morel J, Fromentin J, Blein JP, Simon-Plas F, Elmayan T (2004) Rac regulation of NtrbohD, the oxidase responsible for the oxidative burst in elicited tobacco cell. Plant J 37:282–293

    PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nibau C, Wu HM, Cheung AY (2006) RAC/ROP GTPases: ‘hubs’ for signal integration and diversification in plants. Trends Plant Sci 11:309–315

    Article  PubMed  CAS  Google Scholar 

  • Nishimura MT, Stein M, Hou BH, Vogel JP, Edwards H, Somerville SC (2003) Loss of a callose synthase results in salicylic acid-dependent disease resistance. Science 301:969–972

    Article  PubMed  CAS  Google Scholar 

  • Ono E, Wong HL, Kawasaki T, Hasegawa M, Kodama O, Shimamoto K (2001) Essential role of the small GTPase Rac in disease resistance of rice. Proc Natl Adad Sci USA 98:759–764

    Article  CAS  Google Scholar 

  • Opalski KS, Schultheiss H, Kogel KH, Hückelhoven R (2005) The receptor-like MLO protein and the RAC/ROP family G-protein RACB modulate actin reorganization in barley attacked by the biotrophic powdery mildew fungus Blumeria graminis f.sp. hordei. Plant J 41:291–303

    Article  PubMed  CAS  Google Scholar 

  • Routledge APM, Shelley G, Smith JV, Talbot NJ, Draper J, Mur LAJ (2004) Magnaporthe grisea interactions with the model grass Brachypodium distachyon closely resemble those with rice (Oryza sativa). Mol Plant Pathol 5:253–265

    Article  CAS  Google Scholar 

  • Schiene K, Puhler A, Niehaus K (2000) Transgenic tobacco plants that express an antisense construct derived from a Medicago sativa cDNA encoding a Rac-related small GTP-binding protein fail to develop necrotic lesions upon elicitor infiltration. Mol Gen Genet 263:761–770

    Article  PubMed  CAS  Google Scholar 

  • Schmelzer E (2002) Cell polarization, a crucial process in fungal defence. Trends Plant Sci 7:411–415

    Article  PubMed  CAS  Google Scholar 

  • Schultheiss H, Dechert C, Kogel KH, Hückelhoven R (2002) A small GTP-binding host protein is required for entry of powdery mildew fungus into epidermal cells of barley. Plant Physiol 128:1447–1454

    Article  PubMed  CAS  Google Scholar 

  • Schultheiss H, Dechert C, Kogel KH, Hückelhoven R (2003) Functional analysis of barley RAC/ROP G-protein family members in susceptibility to the powdery mildew fungus. Plant J 36:589–601

    Article  PubMed  CAS  Google Scholar 

  • Schultheiss H, Hensel G, Imani J, Broeders S, Kumlehn J, Kogel KH, Sonnewald U, Hückelhoven R (2005) Ectopic expression of constitutively activated RACB in barley enhances susceptibility to powdery mildew and abiotic stress. Plant Physiol 139:353–362

    Article  PubMed  CAS  Google Scholar 

  • Schultheiss H, Preuss J, Pircher T, Eichmann R, Hückelhoven R (2008) Barley RIC171 interacts with RACB in planta and supports entry of the powdery mildew fungus. Cell Microbiol 10:1815–1826

    Article  PubMed  CAS  Google Scholar 

  • Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localisation of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J 11:1187–1194

    Article  CAS  Google Scholar 

  • Trujillo M, Altschmied L, Schweizer P, Kogel KH, Hückelhoven R (2006) Respiratory burst oxidase homologue A of barley contributes to penetration by the powdery mildew fungus Blumeria graminis f.sp. hordei. J Exp Bot 57:3781–3791

    Article  PubMed  CAS  Google Scholar 

  • Truernit E, Bauby H, Dubreucq B, Grandjean O, Runions J, Barthélémy J, Palauqui JC (2008) High-resolution whole-mount imaging of three-dimensional tissue organization and gene expression enables the study of phloem development and structure in Arabidopsis. Plant Cell 20:1494–1503

    Article  PubMed  CAS  Google Scholar 

  • Vernoud V, Horton AC, Yang Z, Nielsen E (2003) Analysis of the small GTPase gene superfamily of Arabidopsis. Plant Physiol 131:1191–1208

    Article  PubMed  CAS  Google Scholar 

  • Wiberg A (1974) Genetical studies of spontaneous sources of resistance to powdery mildew in barley. Hereditas 77:89–148

    Article  PubMed  CAS  Google Scholar 

  • Winge P, Brembu T, Kristensen R, Bones AM (2000) Genetic structure and evolution of RAC-GTPases in Arabidopsis thaliana. Genetics 156:1959–1971

    PubMed  CAS  Google Scholar 

  • Wong HL, Pinontoan R, Hayashi K, Tabata R, Yaeno T, Hasegawa K, Kojima C, Yoshioka H, Iba K, Kawasaki T, Shimamoto K (2007) Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension. Plant Cell 19:4022–4034

    Article  PubMed  CAS  Google Scholar 

  • Wong HL, Sakamoto T, Kawasaki T, Umemura K, Shimamoto K (2004) Down-regulation of metallothionein, a reactive oxygen scavenger, by the small GTPase OsRac1 in rice. Plant Physiol 135:1447–1456

    Article  PubMed  CAS  Google Scholar 

  • Yang G, Gao P, Zhang H, Huang S, Zheng ZL (2007) A mutation in MRH2 kinesin enhances the root hair tip growth defect caused by constitutively activated ROP2 small GTPase in Arabidopsis. PLoS ONE 2:e1074

    Article  PubMed  CAS  Google Scholar 

  • Zellerhoff N, Jarosch B, Groenewald JZ, Crous PW, Schaffrath U (2006) Nonhost resistance of barley is successfully manifested against Magnaporthe grisea and a closely related Pennisetum-infecting lineage but is overcome by Magnaporthe oryzae. Mol Plant Microbe Interact 19:1014–1022

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Cornelia Marthe (IPK Gatersleben) for technical assistance. This work was funded by the German Research Foundation (HU886/1-3), by BMBF-GABI-Agrotec, by the German Academic Exchange Service, and by the BASF AG.

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Authors and Affiliations

  1. Lehrstuhl für Phytopathologie, Technische Universität München, Am Hochanger 2, 85350, Freising, Germany

    Indira Priyadarshini Pathuri, Ruth Eichmann & Ralph Hückelhoven

  2. Institute for Biology III (Plant Physiology), RWTH Aachen, 52056, Aachen, Germany

    Nina Zellerhoff & Ulrich Schaffrath

  3. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstraße 3, 06466, Gatersleben, Germany

    Götz Hensel & Jochen Kumlehn

  4. Institute of Phytopathology and Applied Zoology, University of Giessen, Heinrich-Buff-Ring 26-32, 35392, Giessen, Germany

    Karl-Heinz Kogel

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  1. Indira Priyadarshini Pathuri
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  2. Nina Zellerhoff
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  3. Ulrich Schaffrath
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  4. Götz Hensel
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  6. Karl-Heinz Kogel
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  7. Ruth Eichmann
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  8. Ralph Hückelhoven
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Correspondence to Ralph Hückelhoven.

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Communicated by A. Feher.

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Pathuri, I.P., Zellerhoff, N., Schaffrath, U. et al. Constitutively activated barley ROPs modulate epidermal cell size, defense reactions and interactions with fungal leaf pathogens. Plant Cell Rep 27, 1877–1887 (2008). https://doi.org/10.1007/s00299-008-0607-9

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  • DOI: https://doi.org/10.1007/s00299-008-0607-9

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