Magnaporthe oryzae effectors MoHEG13 and MoHEG16 interfere with host infection and MoHEG13 counteracts cell death caused by Magnaporthe-NLPs in tobacco
- 763 Downloads
Adapted pathogens are able to modulate cell responses of their hosts most likely due to the activity of secreted effector molecules thereby enabling colonisation by ostensible nonhost pathogens.
It is postulated that host and nonhost pathogens of a given plant species differ in their repertoire of secreted effector molecules that are able to suppress plant resistance. We pursued the strategy of identifying novel effectors of Magnaporthe oryzae, the causal agent of blast disease, by comparing the infection process of closely related host vs. nonhost Magnaporthe species on barley (Hordeum vulgare L.). When both types of pathogen simultaneously attacked the same cell, the nonhost isolate became a successful pathogen possibly due to potent effectors secreted by the host isolate. Microarray studies led to a set of M. oryzae Hypothetical Effector Genes (MoHEGs) which were classified as Early- and LateMoHEGs according to the maximal transcript abundance during colonization of barley. Interestingly, orthologs of these MoHEGs from a nonhost pathogen were similarly regulated when investigated in a host situation, suggesting evolutionary conserved functions. Knockout mutants of MoHEG16 from the group of EarlyMoHEGs were less virulent on barley and microscopic studies revealed an attenuated transition from epidermal to mesophyll colonization. MoHEG13, a LateMoHEG, was shown to antagonize cell death induced by M. oryzae Necrosis-and ethylene-inducing-protein-1 (Nep1)-like proteins in Nicotiana benthamiana. MoHEG13 has a virulence function as a knockout mutant showed attenuated disease progression when inoculated on barley.
KeywordsMagnaporthe oryzae Barley Effector proteins Necrosis
The authors are grateful to Nick Talbot, Lauren Ryder (both University of Exeter, UK) and Holger Deising, Ralf Horbach and Jan-Jorrit Krijger (all Martin-Luther-University Halle-Wittenberg, Germany) for introducing Valerie Mogga into Magnaporthe transformation and for sharing material. Didier Tharreau (CIRAD, Montpellier, France) is kindly acknowledged for providing different Magnaporthe isolates. Jochen Kleemann und Richard O’Connell (both MPIPZ, Cologne, Germany) helped with the N. benthamiana infiltration assay and shared material. We are also grateful to Andrew J. Foster (IBWF, Kaiserslautern, Germany) for providing the pCAMB-HPT vector and to Hiromasa Saitoh (Iwate Biotechnology Research Center, Kitakami, Japan) for sharing Δmoheg13 and wild type isolate INA72. Denise Weidenbach was funded in the framework of the Federal Ministry of Education and Research funding activity ‘Plant Biotechnology for the future, PLANT 2030’ within the project ‘BarleyFortress’.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B (Methodol) 57:289–300Google Scholar
- Catlett NL, Lee DN, Yoder OC, Turgeon BG (2003) Split-marker recombination for efficient targeted deletion of fungal genes. Fungal Genet Newsl 50:9–11Google Scholar
- Cesari S, Thilliez G, Ribot C, Chalvon V, Michel C, Jauneau A, Rivas S, Alaux L, Kanzaki H, Okuyama Y, Morel J-B, Fournier E, Tharreau D, Terauchi R, Kroj T (2013) The rice resistance protein pair RGA4/RGA5 recognizes the Magnaporthe oryzae effectors AVR-Pia and AVR1-CO39 by direct binding. Plant Cell 25:1463–1481CrossRefPubMedPubMedCentralGoogle Scholar
- Chen S, Songkumarn P, Venu RC, Gowda M, Bellizzi M, Hu J, Liu W, Ebbole D, Meyers B, Mitchell T, Wang G-L (2013) Identification and characterization of in planta-expressed secreted effector proteins from Magnaporthe oryzae that induce cell death in rice. Mol Plant Microbe Interact 26:191–202CrossRefPubMedGoogle Scholar
- Dean RA, Talbot NJ, Ebbole DJ, Farman ML, Mitchell TK, Orbach MJ, Thon M, Kulkarni R, Xu J-R, Pan H, Read ND, Lee Y-H, Carbone I, Brown D, Oh YY, Donofrio N, Jeong JS, Soanes DM, Djonovic S, Kolomiets E, Rehmeyer C, Li W, Harding M, Kim S, Lebrun M-H, Bohnert H, Coughlan S, Butler J, Calvo S, Ma L-J, Nicol R, Purcell S, Nusbaum C, Galagan JE, Birren BW (2005) The genome sequence of the rice blast fungus Magnaporthe grisea. Nature 434:980–986CrossRefPubMedGoogle Scholar
- Gentleman R, Carey V, Bates D, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini A, Sawitzki G, Smith C, Smyth G, Tierney L, Yang J, Zhang J (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5:R80CrossRefPubMedPubMedCentralGoogle Scholar
- Gowda M, Venu R, Raghupathy M, Nobuta K, Li H, Wing R, Stahlberg E, Couglan S, Haudenschild C, Dean R, Nahm B-H, Meyers B, Wang G-L (2006) Deep and comparative analysis of the mycelium and appressorium transcriptomes of Magnaporthe grisea using MPSS, RL-SAGE, and oligoarray methods. BMC Genom 7:310CrossRefGoogle Scholar
- Horton P, Park KJ, Obayashi T, Nakai K (2006) Protein subcellular localization prediction with WOLF PSORT. Ser Adv Bioinform 3:39–48Google Scholar
- Kleemann J, Rincon-Rivera LJ, Takahara H, Neumann U, van Themaat EVL, van der Does HC, Hacquard S, Stüber K, Will I, Schmalenbach W, Schmelzer E, O’Connell RJ (2012) Sequential delivery of host-induced virulence effectors by appressoria and intracellular hyphae of the phytopathogen Colletotrichum higginsianum. PLoS Pathog 8:e1002643CrossRefPubMedPubMedCentralGoogle Scholar
- Kunoh H, Kuroda K, Hayashimoto A, Ishizaki H (1986) Induced susceptibility and enhanced resistance at the cellular level in barley coleoptiles. II. Timing and localization of induced susceptibility in a single coleoptile cell and its transfer to an adjacent cell. Can J Bot 64:889–895CrossRefGoogle Scholar
- Loehrer M, Botterweck J, Jahnke J, Mahlmann DM, Gaetgens J, Oldiges M, Horbach R, Deising H, Schaffrath U (2014) In vivo assessment by Mach-Zehnder double-beam interferometry of the invasive force exerted by the Asian soybean rust fungus (Phakopsora pachyrhizi). New Phytol 203:620–631CrossRefPubMedGoogle Scholar
- Mathioni SM, Patel N, Riddick B, Sweigard JA, Czymmek KJ, Caplan JL, Kunjeti SG, Kunjeti S, Raman V, Hillman BI, Kobayashi DY, Donofrio NM (2013) Transcriptomics of the rice blast fungus Magnaporthe oryzae in response to the bacterial antagonist Lysobacter enzymogenes reveals candidate fungal defense response genes. PLoS One 8:e76487CrossRefPubMedPubMedCentralGoogle Scholar
- Motteram J, Küfner I, Deller S, Brunner F, Hammond-Kosack KE, Nürnberger T, Rudd JJ (2009) Molecular characterization and functional analysis of MgNLP, the sole NPP1 domain-containing protein, from the fungal wheat leaf pathogen Mycosphaerella graminicola. Mol Plant Microbe Interact 22:790–799CrossRefPubMedGoogle Scholar
- Qutob D, Kemmerling B, Brunner F, Küfner I, Engelhardt S, Gust AA, Luberacki B, Seitz HU, Stahl D, Rauhut T, Glawischnig E, Schween G, Lacombe B, Watanabe N, Lam E, Schlichting R, Scheel D, Nau K, Dodt G, Hubert D, Gijzen M, Nürnberger T (2006) Phytotoxicity and innate immune responses induced by Nep1-like proteins. Plant Cell 18:3721–3744CrossRefPubMedPubMedCentralGoogle Scholar
- R Development Core Team (2009). http://www.R-project.org. Accessed 15 July 2009
- Saitoh H, Fujisawa S, Mitsuoka C, Ito A, Hirabuchi A, Ikeda K, Irieda H, Yoshino K, Yoshida K, Matsumura H, Tosa Y, Win J, Kamoun S, Takano Y, Terauchi R (2012) Large-scale gene disruption in Magnaporthe oryzae identifies MC69, a secreted protein required for infection by monocot and dicot fungal pathogens. PLoS Pathog 8:e1002711CrossRefPubMedPubMedCentralGoogle Scholar
- Smyth GK (2004) Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 3:1–25Google Scholar
- Tucker S, Orbach M (2007) Agrobacterium-mediated transformation to create an insertion library in Magnaporthe grisea. In: Ronald P (ed) Plant-pathogen interactions. Humana Press, New York, pp 57–68Google Scholar
- Whisson SC, Boevink PC, Moleleki L, Avrova AO, Morales JG, Gilroy EM, Armstrong MR, Grouffaud S, van West P, Chapman S, Hein I, Toth IK, Pritchard L, Birch PRJ (2007) A translocation signal for delivery of oomycete effector proteins into host plant cells. Nature 450:115–118CrossRefPubMedGoogle Scholar
- Wu J, Kou Y, Bao J, Li Y, Tang M, Zhu X, Ponaya A, Xiao G, Li J, Li C, Song M-Y, Cumagun CJR, Deng Q, Lu G, Jeon JS, Naqvi NI, Zhou B (2015) Comparative genomics identifies the Magnaporthe oryzae avirulence effector AvrPi9 that triggers Pi9-mediated blast resistance in rice. New Phytologist 206:1463–1475CrossRefPubMedGoogle 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 19:1014–1022CrossRefPubMedGoogle Scholar
- Zhang H, Li D, Wang M, Liu J, Teng W, Cheng B, Huang Q, Wang M, Song W, Dong S, Zheng X, Zhang Z (2012) The Nicotiana benthamiana mitogen-activated protein kinase cascade and WRKY transcription factor participate in Nep1Mo-triggered plant responses. Mol Plant Microbe Interact 25:1639–1653CrossRefPubMedGoogle Scholar