Summary:
Secreted effectors play crucial roles during the establishment of the biotrophic interaction between Ustilago maydis and maize. In a previous study (Schipper, 2009) it had been demonstrated that a deletion of the stp1 effector gene resulted in a complete loss of virulence symptoms in maize infection and that such mutants elicited a hypersensitive response. This distinguishes stp1 from most other secreted effectors that are either dispensable for pathogenicity or have only a minor effect on virulence. This study focuses
on the functional analysis of Stp1.
A mutational analysis showed that the conserved N- and C-terminal domains of Stp1 can be separately expressed but are both required for Stp1 protein function. The long central variable domain was demonstrated to be dispensable yet may promote fungal growth in vascular bundles. stp1 homologs from closely related smut fungi of U. maydis could replace stp1 in U. maydis, indicating a conserved function. Stp1Δ136-432 lacking the central domain could be purified to homogeneity and was stable, while the isolated C-terminal domain, Stp1433-515, was unstable after purification. This could suggest that N- and Cterminal domains of Stp1 stabilize each other. Stp1-HA expressed by U. maydis was detected in the nucleus of plant cells by immunolocalization suggesting that Stp1 may suppress plant defense responses by affecting the transcription of respective genes.
Both cytoplasmic and apoplastic maize proteins were identified as interaction partners of Stp1 by yeast two-hybrid assays using Stp1Δ136-432 as bait, suggesting that Stp1 may be an effector with both apoplastic and cytoplasmic functions The C-terminus of Stp1 as well as Stp1Δ136-432 could inhibit the activity of a maize extracellular cysteine protease, Sip3, which was identified as one of the apoplastic interaction partners. The interactions between Stp1
and the cytoplasmic interactors Sip9, a cell number regulator 8, Sip16, a CCR4-NOT transcription complex subunit, Sip19, a serine/threonine-protein kinase and Sip21, a VIP2 protein were verified with full-length cDNA but await to be confirmed by other techniques.
RNA-Seq analysis demonstrated that several early defense response genes are not induced by stp1 mutants expressing the N-terminus of Stp1 while stp1 mutants expressing the Cterminus of Stp1 triggered even stronger plant defense responses than stp1 mutants during colonization. This suggests that N- and C-terminal domains of Stp1 have distinct functions.
Bibliographie / References
- Tian, M., E. Huitema, L. Da Cunha, T. Torto-Alalibo & S. Kamoun, (2004) A Kazal-like extracellular serine protease inhibitor from Phytophthora infestans targets the tomato pathogenesis-related protease P69B. J Biol Chem 279: 26370-26377.
- Rawlings, N. D., A. J. Barrett & A. Bateman, (2012) MEROPS: the database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Research 40: D343-D350.
- Dubery, I. A., N. M. Sanabria & J. C. Huang, (2012) Nonself perception in plant innate immunity. Adv Exp Med Biol 738: 79-107.
- Kamper, J., (2004) A PCR-based system for highly efficient generation of gene replacement mutants in Ustilago maydis. Mol Genet Genomics 271: 103-110.
- Perez-Martin, J., S. Castillo-Lluva, C. Sgarlata, I. Flor-Parra, N. Mielnichuk, J. Torreblanca & N. Carbo, (2006) Pathocycles: Ustilago maydis as a model to study the relationships between cell cycle and virulence in pathogenic fungi. Mol Genet Genomics 276: 211-229.
- Banuett, F., (1992) Ustilago maydis, the delightful blight. Trends Genet 8: 174-180.
- Munis, M. F., L. Tu, F. Deng, J. Tan, L. Xu, S. Xu, L. Long & X. Zhang, (2010) A thaumatin-like protein gene involved in cotton fiber secondary cell wall development enhances resistance against Verticillium dahliae and other stresses in transgenic tobacco. Biochem Biophys Res Commun 393: 38-44.
- Panstruga, R., J. E. Parker & P. Schulze-Lefert, (2009) SnapShot: Plant immune response pathways. Cell 136: 978 e971-973.
- Bhattacharjee, S., R. V. Stahelin, K. D. Speicher, D. W. Speicher & K. Haldar, (2012) Endoplasmic reticulum PI(3)P lipid binding targets malaria proteins to the host cell. Cell 148: 201-212.
- Kloppholz, S., H. Kuhn & N. Requena, (2011) A secreted fungal effector of Glomus intraradices promotes symbiotic biotrophy. Current biology : CB 21: 1204-1209.
- Holliday, R., (2004) Early studies on recombination and DNA repair in Ustilago maydis. DNA Repair (Amst) 3: 671-682.
- Hann, D. R., S. Gimenez-Ibanez & J. P. Rathjen, (2010) Bacterial virulence effectors and their activities. Curr Opin Plant Biol 13: 388-393.
- Stassen, J. H. & G. Van den Ackerveken, (2011) How do oomycete effectors interfere with plant life? Curr Opin Plant Biol 14: 407-414.
- Feng, F. & J. M. Zhou, (2012) Plant-bacterial pathogen interactions mediated by type III effectors. Curr Opin Plant Biol.
- Rafiqi, M., J. G. Ellis, V. A. Ludowici, A. R. Hardham & P. N. Dodds, (2012) Challenges and progress towards understanding the role of effectors in plant-fungal interactions. Curr Opin Plant Biol.
- Collier, S. M. & P. Moffett, (2009) NB-LRRs work a "bait and switch" on pathogens. Trends in plant science 14: 521-529.
- Nunez-Corcuera, B., I. Serafimidis, E. Arias-Palomo, A. Rivera-Calzada & T. Suarez, (2008) A new protein carrying an NmrA-like domain is required for cell differentiation and development in Dictyostelium discoideum. Dev Biol 321: 331- 342.
- Kunkel, B. N. & D. M. Brooks, (2002) Cross talk between signaling pathways in pathogen defense. Curr Opin Plant Biol 5: 325-331.
- Svensson, B., I. Svendsen, P. Hojrup, P. Roepstorff, S. Ludvigsen & F. M. Poulsen, (1992) Primary structure of barwin: a barley seed protein closely related to the C-terminal domain of proteins encoded by wound-induced plant genes. Biochemistry 31: 8767- 8770.
- Galea, C. A., Y. Wang, S. G. Sivakolundu & R. W. Kriwacki, (2008) Regulation of cell division by intrinsically unstructured proteins: intrinsic flexibility, modularity, and signaling conduits. Biochemistry 47: 7598-7609.
- Sauer, M., T. Paciorek, E. Benkova & J. Friml, (2006) Immunocytochemical techniques for whole-mount in situ protein localization in plants. Nat Protoc 1: 98-103.
- Dodds, P. N. & J. P. Rathjen, (2010) Plant immunity: towards an integrated view of plant- pathogen interactions. Nature reviews. Genetics 11: 539-548.
- Loubradou, G., A. Brachmann, M. Feldbrugge & R. Kahmann, (2001) A homologue of the transcriptional repressor Ssn6p antagonizes cAMP signalling in Ustilago maydis. Mol Microbiol 40: 719-730.
- Bozkurt, T. O., S. Schornack, J. Win, T. Shindo, M. Ilyas, R. Oliva, L. M. Cano, A. M. E. Jones, E. Huitema, R. A. L. van der Hoorn & S. Kamoun, (2011) Phytophthora infestans effector AVRblb2 prevents secretion of a plant immune protease at the haustorial interface. Proc Natl Acad Sci U S A 108: 20832-20837.
- van den Burg, H. A., N. Westerink, K. J. Francoijs, R. Roth, E. Woestenenk, S. Boeren, P. J. de Wit, M. H. Joosten & J. Vervoort, (2003) Natural disulfide bond-disrupted mutants of AVR4 of the tomato pathogen Cladosporium fulvum are sensitive to proteolysis, circumvent Cf-4-mediated resistance, but retain their chitin binding ability. J Biol Chem 278: 27340-27346.
- de las Mercedes Dana, M., J. A. Pintor-Toro & B. Cubero, (2006) Transgenic tobacco plants overexpressing chitinases of fungal origin show enhanced resistance to biotic and abiotic stress agents. Plant Physiol 142: 722-730.
- Martinez, M., I. Cambra, L. Carrillo, M. Diaz-Mendoza & I. Diaz, (2009) Characterization of the entire cystatin gene family in barley and their target cathepsin L-like cysteine-proteases, partners in the hordein mobilization during seed germination. Plant Physiol 151: 1531-1545.
- Guo, M., M. A. Rupe, J. A. Dieter, J. Zou, D. Spielbauer, K. E. Duncan, R. J. Howard, Z. Hou & C. R. Simmons, (2010) Cell Number Regulator1 affects plant and organ size in maize: implications for crop yield enhancement and heterosis. Plant Cell 22: 1057-1073.
- Mowbray, S. L., L. T. Elfstrom, K. M. Ahlgren, C. E. Andersson & M. Widersten, (2006) X-ray structure of potato epoxide hydrolase sheds light on substrate specificity in plant enzymes. Protein Sci 15: 1628-1637.
- Basse, C. W. & G. Steinberg, (2004) Ustilago maydis, model system for analysis of the molecular basis of fungal pathogenicity. Mol Plant Pathol 5: 83-92.
- Staskawicz, B., (2009) First insights into the genes that control plant-bacterial interactions. Mol Plant Pathol 10: 719-720.
- Doehlemann, G., S. Reissmann, D. Assmann, M. Fleckenstein & R. Kahmann, (2011) Two linked genes encoding a secreted effector and a membrane protein are essential for Ustilago maydis-induced tumour formation. Mol Microbiol 81: 751-766.
- Sarowar, S., H. W. Oh, H. S. Cho, K. H. Baek, E. S. Seong, Y. H. Joung, G. J. Choi, S. Lee & D. Choi, (2007) Capsicum annuum CCR4-associated factor CaCAF1 is necessary for plant development and defence response. Plant J 51: 792-802.
- Lee, M. W., J. Jelenska & J. T. Greenberg, (2008) Arabidopsis proteins important for modulating defense responses to Pseudomonas syringae that secrete HopW1-1. Plant Journal 54: 452-465.
- Doehlemann, G., R. Wahl, R. J. Horst, L. M. Voll, B. Usadel, F. Poree, M. Stitt, J. Pons- Kuhnemann, U. Sonnewald, R. Kahmann & J. Kamper, (2008) Reprogramming a maize plant: transcriptional and metabolic changes induced by the fungal biotroph Ustilago maydis. Plant J 56: 181-195.
- Shimada, Y., S. Ichinose, A. Sadr, M. F. Burrow & J. Tagami, (2009) Localization of matrix metalloproteinases (MMPs-2, 8, 9 and 20) in normal and carious dentine. Australian dental journal 54: 347-354.
- Hardcastle, T. J. & K. A. Kelly, (2010) baySeq: empirical Bayesian methods for identifying differential expression in sequence count data. BMC bioinformatics 11: 422.
- Chung, C. L., J. M. Longfellow, E. K. Walsh, Z. Kerdieh, G. Van Esbroeck, P. Balint-Kurti & R. J. Nelson, (2010) Resistance loci affecting distinct stages of fungal pathogenesis: use of introgression lines for QTL mapping and characterization in the maize -Setosphaeria turcica pathosystem. Bmc Plant Biol 10.
- Gao, D., J. Kim, H. Kim, T. L. Phang, H. Selby, A. C. Tan & T. Tong, (2010) A survey of statistical software for analysing RNA-seq data. Human genomics 5: 56-60.
- Robinson, M. D. & A. Oshlack, (2010) A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol 11: R25.
- Kvitko, B. H., D. H. Park, A. C. Velasquez, C. F. Wei, A. B. Russell, G. B. Martin, D. J. Schneider & A. Collmer, (2009) Deletions in the repertoire of Pseudomonas syringae pv. tomato DC3000 type III secretion effector genes reveal functional overlap among effectors. PLoS Pathog 5: e1000388.
- Kvam, V. M., P. Liu & Y. Si, (2012) A comparison of statistical methods for detecting differentially expressed genes from RNA-seq data. American journal of botany 99: 248-256.
- Stergiopoulos, I. & P. J. de Wit, (2009) Fungal effector proteins. Annu Rev Phytopathol 47: 233-263.
- de Jonge, R., M. D. Bolton & B. P. Thomma, (2011) How filamentous pathogens co-opt plants: the ins and outs of fungal effectors. Curr Opin Plant Biol 14: 400-406.
- Djamei, A., K. Schipper, F. Rabe, A. Ghosh, V. Vincon, J. Kahnt, S. Osorio, T. Tohge, A. R. Fernie, I. Feussner, K. Feussner, P. Meinicke, Y. D. Stierhof, H. Schwarz, B. Macek, M. Mann & R. Kahmann, (2011) Metabolic priming by a secreted fungal effector. Nature 478: 395-398.
- Tian, M., J. Win, J. Song, R. van der Hoorn, E. van der Knaap & S. Kamoun, (2007) A Phytophthora infestans cystatin-like protein targets a novel tomato papain-like apoplastic protease. Plant Physiol 143: 364-377.
- Koeck, M., A. R. Hardham & P. N. Dodds, (2011) The role of effectors of biotrophic and hemibiotrophic fungi in infection. Cellular microbiology 13: 1849-1857.
- Khang, C. H., R. Berruyer, M. C. Giraldo, P. Kankanala, S. Y. Park, K. Czymmek, S. Kang & B. Valent, (2010) Translocation of Magnaporthe oryzae effectors into rice cells and their subsequent cell-to-cell movement. Plant Cell 22: 1388-1403.
- Sambrook, J., E. F. Fritsch & T. Maniatis., (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York, USA.
- Bennetzen, J. L. H., Sarah C. (Eds.), (2009) Handbook of Maize, Genetics and Genomics. XII, 800 p. 894 illus.
- van der Linde, K., C. Hemetsberger, C. Kastner, F. Kaschani, R. A. van der Hoorn, J. Kumlehn & G. Doehlemann, (2012) A maize cystatin suppresses host immunity by inhibiting apoplastic cysteine proteases. Plant Cell 24: 1285-1300.
- Voinnet, O., S. Rivas, P. Mestre & D. Baulcombe, (2003) An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 33: 949-956.
- Peng, Y. L., Y. Shirano, H. Ohta, T. Hibino, K. Tanaka & D. Shibata, (1994) A Novel Lipoxygenase from Rice -Primary Structure and Specific Expression Upon Incompatible Infection with Rice Blast Fungus. Journal of Biological Chemistry 269: 3755-3761.
- Bradford, M. M., (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
- Cornelis, G. R. & F. Van Gijsegem, (2000) Assembly and function of type III secretory systems. Annual review of microbiology 54: 735-774.
- Pritchard, L. & P. Birch, (2011) A systems biology perspective on plant-microbe interactions: biochemical and structural targets of pathogen effectors. Plant science : an international journal of experimental plant biology 180: 584-603.
- Whisson, S. C., P. C. Boevink, L. Moleleki, A. O. Avrova, J. G. Morales, E. M. Gilroy, M. R. Armstrong, S. Grouffaud, P. van West, S. Chapman, I. Hein, I. K. Toth, L. Pritchard & P. R. Birch, (2007) A translocation signal for delivery of oomycete effector proteins into host plant cells. Nature 450: 115-118.
- References Abramovitch, R. B. & G. B. Martin, (2005) AvrPtoB: a bacterial type III effector that both elicits and suppresses programmed cell death associated with plant immunity. FEMS microbiology letters 245: 1-8.
- Schipper, K., (2009) Charakterisierung eines Ustilago maydis Genclusters, das für drei neuartige sekretierte Effektoren kodiert. PhD thesis.
- Rooney, H. C., J. W. Van't Klooster, R. A. van der Hoorn, M. H. Joosten, J. D. Jones & P. J. de Wit, (2005) Cladosporium Avr2 inhibits tomato Rcr3 protease required for Cf- 2-dependent disease resistance. Science 308: 1783-1786.
- Otto, H. H. & T. Schirmeister, (1997) Cysteine proteases and their inhibitors. Chem Rev 97: 133-171.
- Mauch, F. & R. Dudler, (1993) Differential induction of distinct glutathione-S-transferases of wheat by xenobiotics and by pathogen attack. Plant Physiol 102: 1193-1201.
- Rehmany, A. P., A. Gordon, L. E. Rose, R. L. Allen, M. R. Armstrong, S. C. Whisson, S. Kamoun, B. M. Tyler, P. R. Birch & J. L. Beynon, (2005) Differential recognition of highly divergent downy mildew avirulence gene alleles by RPP1 resistance genes from two Arabidopsis lines. Plant Cell 17: 1839-1850.
- Robinson, M. D., D. J. McCarthy & G. K. Smyth, (2010) edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26: 139-140.
- Kale, S. D., B. Gu, D. G. Capelluto, D. Dou, E. Feldman, A. Rumore, F. D. Arredondo, R. Hanlon, I. Fudal, T. Rouxel, C. B. Lawrence, W. Shan & B. M. Tyler, (2010) External lipid PI3P mediates entry of eukaryotic pathogen effectors into plant and animal host cells. Cell 142: 284-295.
- Vollmeister, E., K. Schipper, S. Baumann, C. Haag, T. Pohlmann, J. Stock & M. Feldbrugge, (2012) Fungal development of the plant pathogen Ustilago maydis. FEMS Microbiol Rev 36: 59-77.
- Dixon, M. S., C. Golstein, C. M. Thomas, E. A. van Der Biezen & J. D. Jones, (2000) Genetic complexity of pathogen perception by plants: the example of Rcr3, a tomato gene required specifically by Cf-2. Proc Natl Acad Sci U S A 97: 8807- 8814.
- Laurie, J. D., S. Ali, R. Linning, G. Mannhaupt, P. Wong, U. Guldener, M. Munsterkotter, R. Moore, R. Kahmann, G. Bakkeren & J. Schirawski, (2012) Genome comparison of barley and maize smut fungi reveals targeted loss of RNA silencing components and species-specific presence of transposable elements. Plant Cell 24: 1733-1745.
- Philipps-Universität Marburg Max-Planck-Institute for Terrestrial Microbiology Marburg/Lahn, Germany PhD thesis: The role of Stp1, a secreted effector, in the biotrophic interaction of Ustilago maydis and its host plant maize Jul 2005–Jan 2008
- Jefferson, R. A., T. A. Kavanagh & M. W. Bevan, (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901- 3907.
- Chisholm, S. T., G. Coaker, B. Day & B. J. Staskawicz, (2006) Host-microbe interactions: shaping the evolution of the plant immune response. Cell 124: 803-814.
- de Wit, P. J., (2007) How plants recognize pathogens and defend themselves. Cellular and molecular life sciences : CMLS 64: 2726-2732.
- Jones, H. D., S. Kurup, N. C. B. Peters & M. J. Holdsworth, (2000) Identification and analysis of proteins that interact with the Avena fatua homologue of the maize transcription factor VIVIPAROUS 1. Plant Journal 21: 133-142.
- Kemen, E., A. C. Kemen, M. Rafiqi, U. Hempel, K. Mendgen, M. Hahn & R. T. Voegele, (2005) Identification of a protein from rust fungi transferred from haustoria into infected plant cells. Molecular plant-microbe interactions : MPMI 18: 1130-1139.
- Hanahan, D., (1985) In DNA Cloning -a Practical Approach. (Glover, D.M., ed.). IRL Press, McLean, Virginia. 1: 109.
- Ortiz-Castellanos, W. Li, P. Sanchez-Alonso, P. H. Schreier, I. Hauser-Hahn, M. Vaupel, E. Koopmann, G. Friedrich, H. Voss, T. Schluter, J. Margolis, D. Platt, C. Swimmer, A. Gnirke, F. Chen, V. Vysotskaia, G. Mannhaupt, U. Guldener, M. Munsterkotter, D. Haase, M. Oesterheld, H. W. Mewes, E. W. Mauceli, D. DeCaprio, C. M. Wade, J. Butler, S. Young, D. B. Jaffe, S. Calvo, C. Nusbaum, J. Galagan & B. W. Birren, (2006) Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis. Nature 444: 97-101.
- Rafiqi, M., P. H. Gan, M. Ravensdale, G. J. Lawrence, J. G. Ellis, D. A. Jones, A. R. Hardham & P. N. Dodds, (2010) Internalization of flax rust avirulence proteins into flax and tobacco cells can occur in the absence of the pathogen. Plant Cell 22: 2017-2032.
- Kunitz, M. & J. H. Northrop, (1936) Isolation from Beef Pancreas of Crystalline Trypsinogen, Trypsin, a Trypsin Inhibitor, and an Inhibitor-Trypsin Compound. The Journal of general physiology 19: 991-1007.
- Burke, D., D. Dawson & T. Stearns, (2000) Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual. Cold Spring Harbor Laboratory Press, New York.
- Rose, J. K., K. S. Ham, A. G. Darvill & P. Albersheim, (2002) Molecular cloning and characterization of glucanase inhibitor proteins: coevolution of a counterdefense mechanism by plant pathogens. Plant Cell 14: 1329-1345.
- Food and Agriculture Organization of the United Nations, S. D., Statistics Division, (2010) Fradin, E. F. & B. P. H. J. Thomma, (2006) Physiology and molecular aspects of Verticillium wilt diseases caused by V-dahliae and V-albo-atrum. Molecular Plant Pathology 7: 71-86.
- Takken, F. L. W. & M. H. A. J. Joosten, (2000) Plant resistance genes: their structure, function and evolution. Eur J Plant Pathol 106: 699-713.
- Fontana, A., P. P. de Laureto, B. Spolaore, E. Frare, P. Picotti & M. Zambonin, (2004) Probing protein structure by limited proteolysis. Acta Biochim Pol 51: 299-321.
- Ellis, J. G., M. Rafiqi, P. Gan, A. Chakrabarti & P. N. Dodds, (2009) Recent progress in discovery and functional analysis of effector proteins of fungal and oomycete plant pathogens. Curr Opin Plant Biol 12: 399-405.
- Bonas, U. & G. Van den Ackervaken, (1997) Recognition of bacterial avirulence proteins occurs inside the plant cell: a general phenomenon in resistance to bacterial diseases? Plant J 12: 1-7.
- Frederick, R. D., R. L. Thilmony, G. Sessa & G. B. Martin, (1998) Recognition specificity for the bacterial avirulence protein AvrPto is determined by Thr-204 in the activation loop of the tomato Pto kinase. Mol Cell 2: 241-245.
- Dou, D., S. D. Kale, X. Wang, R. H. Jiang, N. A. Bruce, F. D. Arredondo, X. Zhang & B. M. Tyler, (2008) RXLR-mediated entry of Phytophthora sojae effector Avr1b into soybean cells does not require pathogen-encoded machinery. Plant Cell 20: 1930- 1947.
- Bull, J., F. Mauch, C. Hertig, G. Rebmann & R. Dudler, (1992) Sequence and Expression of a Wheat Gene That Encodes a Novel Protein Associated with Pathogen Defense. Mol Plant Microbe In 5: 516-519.
- Meshi & M. Nishimura, (2012) Surface alpha-1,3-Glucan Facilitates Fungal Stealth Infection by Interfering with Innate Immunity in Plants. PLoS Pathog 8: e1002882.
- Takken, F. & M. Rep, (2010) The arms race between tomato and Fusarium oxysporum. Mol Plant Pathol 11: 309-314.
- Hann, D. R. & J. P. Rathjen, (2010) The long and winding road: virulence effector proteins of plant pathogenic bacteria. Cellular and molecular life sciences : CMLS 67: 3425- 3434.
- Harris, L. J., A. Saparno, A. Johnston, S. Prisic, M. Xu, S. Allard, A. Kathiresan, T. Ouellet & R. J. Peters, (2005) The maize An2 gene is induced by Fusarium attack and encodes an ent-copalyl diphosphate synthase. Plant Molecular Biology 59: 881-894.
- Jones, J. D. & J. L. Dangl, (2006) The plant immune system. Nature 444: 323-329.
- Schoepfer, R., (1993) The pRSET family of T7 promoter expression vectors for Escherichia coli. Gene 124: 83-85.
- Dean, R., J. A. Van Kan, Z. A. Pretorius, K. E. Hammond-Kosack, A. Di Pietro, P. D. Spanu, J. J. Rudd, M. Dickman, R. Kahmann, J. Ellis & G. D. Foster, (2012) The Top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol 13: 414- 430.
- Leung-Toung, R., W. R. Li, T. F. Tam & K. Karimian, (2002) Thiol-dependent enzymes and their inhibitors: A review. Curr Med Chem 9: 979-1002.
- Schagger, H., (2006) Tricine-SDS-PAGE. Nat Protoc 1: 16-22.
- Brefort, T., G. Doehlemann, A. Mendoza-Mendoza, S. Reissmann, A. Djamei & R. Kahmann, (2009) Ustilago maydis as a Pathogen. Annu Rev Phytopathol 47: 423- 445.
- Bolker, M., (2001) Ustilago maydis--a valuable model system for the study of fungal dimorphism and virulence. Microbiology 147: 1395-1401.
- Kahmann, R. S., G.; Basse, C.; Feldbrügge, M.; Kämper, J. , (2000) Ustilago maydis, the causative agent of corn smut disease. Fungal pathology 347-371
- Munkvold, K. R. & G. B. Martin, (2009) Advances in experimental methods for the elucidation of Pseudomonas syringae effector function with a focus on AvrPtoB. Mol Plant Pathol 10: 777-793.
- Aichinger, C., K. Hansson, H. Eichhorn, F. Lessing, G. Mannhaupt, W. Mewes & R. Kahmann, (2003) Identification of plant-regulated genes in Ustilago maydis by enhancer-trapping mutagenesis. Mol Genet Genomics 270: 303-314.
- Schirawski, J., G. Mannhaupt, K. Munch, T. Brefort, K. Schipper, G. Doehlemann, M. Di Stasio, N. Rossel, A. Mendoza-Mendoza, D. Pester, O. Muller, B. Winterberg, E. Meyer, H. Ghareeb, T. Wollenberg, M. Munsterkotter, P. Wong, M. Walter, E. Stukenbrock, U. Guldener & R. Kahmann, (2010) Pathogenicity determinants in smut fungi revealed by genome comparison. Science 330: 1546-1548.
- Skibbe, D. S., G. Doehlemann, J. Fernandes & V. Walbot, (2010) Maize tumors caused by Ustilago maydis require organ-specific genes in host and pathogen. Science 328: 89- 92.
- Mortazavi, A., B. A. Williams, K. McCue, L. Schaeffer & B. Wold, (2008) Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nature methods 5: 621-628.
- van der Hoorn, R. A. & S. Kamoun, (2008) From Guard to Decoy: a new model for perception of plant pathogen effectors. Plant Cell 20: 2009-2017.
- Doehlemann, G., K. van der Linde, D. Assmann, D. Schwammbach, A. Hof, A. Mohanty, D. Jackson & R. Kahmann, (2009) Pep1, a secreted effector protein of Ustilago maydis, is required for successful invasion of plant cells. PLoS Pathog 5: e1000290.
- Song, J., J. Win, M. Tian, S. Schornack, F. Kaschani, M. Ilyas, R. A. van der Hoorn & S. Kamoun, (2009) Apoplastic effectors secreted by two unrelated eukaryotic plant pathogens target the tomato defense protease Rcr3. Proc Natl Acad Sci U S A 106: 1654-1659.
- Shan, L., P. He, J. Li, A. Heese, S. C. Peck, T. Nurnberger, G. B. Martin & J. Sheen, (2008) Bacterial effectors target the common signaling partner BAK1 to disrupt multiple MAMP receptor-signaling complexes and impede plant immunity. Cell host & microbe 4: 17-27.
- Hemetsberger, C., C. Herrberger, B. Zechmann, M. Hillmer & G. Doehlemann, (2012) The Ustilago maydis effector Pep1 suppresses plant immunity by inhibition of host peroxidase activity. PLoS Pathog 8: e1002684.
- Bölker, M., Genin, S., Lehmler, C. and Kahmann, R, (1995) Genetic regulation of mating and dimorphism in Ustilago maydis. Can. J. Bot 73: 320-325
- Kankanala, P., K. Czymmek & B. Valent, (2007) Roles for rice membrane dynamics and plasmodesmata during biotrophic invasion by the blast fungus. Plant Cell 19: 706- 724.
- Haseloff, J., K. R. Siemering, D. C. Prasher & S. Hodge, (1997) Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc Natl Acad Sci U S A 94: 2122-2127.
- Benz, B. F., (2001) Archaeological evidence of teosinte domestication from Guila Naquitz, Oaxaca. Proc Natl Acad Sci U S A 98: 2104-2106.
- Schulz, B., F. Banuett, M. Dahl, R. Schlesinger, W. Schafer, T. Martin, I. Herskowitz & R. Kahmann, (1990) The b alleles of U. maydis, whose combinations program pathogenic development, code for polypeptides containing a homeodomain-related motif. Cell 60: 295-306.
- Hoffman, C. S. & F. Winston, (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57: 267- 272.
- Kasparovsky, T., J. P. Blein & V. Mikes, (2004) Ergosterol elicits oxidative burst in tobacco cells via phospholipase A2 and protein kinase C signal pathway. Plant physiology and biochemistry : PPB / Societe francaise de physiologie vegetale 42: 429-435.
- Zipfel, C., (2008) Pattern-recognition receptors in plant innate immunity. Current opinion in immunology 20: 10-16. Curriculum Vitae Personal information Name Liang Liang Date of birth Dec 1979