Puccinia striiformis f. sp. tritici (Pst) is an obligate biotrophic fungus that causes the destructive wheat stripe rust disease worldwide. Due to the lack of reliable transformation and gene disruption method, knowledge about the function of Pst genes involved in pathogenesis is limited. Mitogen-activated protein kinase (MAPK) genes have been shown in a number of plant pathogenic fungi to play critical roles in regulating various infection processes. In the present study, we identified and characterized the first MAPK gene PsMAPK1 in Pst. Phylogenetic analysis indicated that PsMAPK1 is a YERK1 MAP kinase belonging to the Fus3/Kss1 class. Single nucleotide polymerphisms (SNPs) and insertion/deletion were detected in the coding region of PsMAPK1 among six Pst isolates. Real-time RT-PCR analyses revealed that PsMAPK1 expression was induced at early infection stages and peaked during haustorium formation. When expressed in Fusarium graminearum, PsMAPK1 partially rescued the map1 mutant in vegetative growth and pathogenicity. It also partially complemented the defects of the Magnaporthe oryzae pmk1 mutant in appressorium formation and plant infection. These results suggest that F. graminearum and M. oryzae can be used as surrogate systems for functional analysis of well-conserved Pst genes and PsMAPK1 may play a role in the regulation of plant penetration and infectious growth in Pst.
References
[1]
Zhao XH, Mehrabi R, Xu JR (2007) Mitogen-activated protein kinase pathways and fungal pathogenesis. Eukaryot Cell 6: 1701–1714.
[2]
Kultz D (1998) Phylogenetic and functional classification of mitogen- and stress-activated protein kinases. J Mol Evol 46: 571–588.
[3]
Saito H, Tatebayashi K (2004) Regulation of the osmoregulatory HOG MAPK cascade in yeast. J Biochem 136: 267–272.
[4]
Xu JR (2000) MAP kinases in fungal pathogens. Fungal Genet Biol 31: 137–152.
[5]
Liu WD, Zhou XY, Li GT, Li L, Kong LG, et al. (2011) Multiple plant surface signals are sensed by different mechanisms in the rice blast fungus for appressorium formation. PLoS Pathog 7: e1001261.
[6]
Good M, Tang G, Singleton J, Remenyi A, Lim WA (2009) The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. Cell 136: 1085–1097.
[7]
Rispail N, Soanes DM, Ant C, Czajkowski R, Grunler A, et al. (2009) Comparative genomics of MAP kinase and calcium-calcineurin signalling components in plant and human pathogenic fungi. Fungal Genet Biol 46: 287–298.
[8]
Xu JR, Hamer JE (1996) MAP kinase and cAMP signaling regulate infection structure formation and pathogenic growth in the rice blast fungus Magnaporthe grisea. Genes Dev 10: 2696–2706.
[9]
Jenczmionka NJ, Maier FJ, Losch AP, Schafer W (2003) Mating, conidiation and pathogenicity of Fusarium graminearum, the main causal agent of the head-blight disease of wheat, are regulated by the MAP kinase GPMK1. Curr Genet 43: 87–95.
[10]
Urban M, Mott E, Farley T, Hammond-Kosack K (2003) The Fusarium graminearum MAP1 gene is essential for pathogenicity and development of perithecia. Mol Plant Path 4: 347–359.
[11]
Basse CW, Steinberg G (2004) Ustilago maydis, model system for analysis of the molecular basis of fungal pathogenicity. Mol Plant Patho 5: 83–92.
[12]
Kahmann R, Kamper J (2004) Ustilago maydis: how its biology relates to pathogenic development. New Phytol 164: 31–42.
[13]
Brachmann A, Schirawski J, Muller P, Kahmann R (2003) An unusual MAP kinase is required for efficient penetration of the plant surface by Ustilago maydis. EMBO J 22: 2199–2210.
[14]
Monge RA, Roman E, Nombela C, Pla J (2006) The MAP kinase signal transduction network in Candida albicans. Microbiology 152: 905–912.
[15]
Davidson RC, Nicholls CB, Cox GM, Perfect JR, Heitman J (2003) A MAP kinase cascade composed of cell type specific and non-specific elements controls mating and differentiation of the fungal pathogen Cryptococcus neoformans. Mol Microbio 49: 469–485.
[16]
Petersen RH (1974) The rust fungus life cycle. Bot Rev 40: 453–513.
[17]
Szabo LJ, Bushnell WR (2001) Hidden robbers: the role of fungal haustoria in parasitism of plants. Proc Natl Acad Sci U S A 98: 7654–7655.
[18]
Voegele RT, Mendgen K (2003) Rust haustoria: nutrient uptake and beyond. New Phytol 159: 93–100.
[19]
Voegele RT, Struck C, Hahn M, Mendgen K (2001) The role of haustoria in sugar supply during infection of broad bean by the rust fungus Uromyces fabae. Proc Natl Acad Sci U S A 98: 8133–8138.
[20]
Wang CF, Huang LL, Buchenauer H, Han QM, Zhang HC, et al. (2007) Histochemical studies on the accumulation of reactive oxygen species (O2？ and H2O2) in the incompatible and compatible interaction of wheat-Puccinia striiformis f. sp. tritici. Physiol Mol Plant Pathol 71: 230–239.
[21]
Hu GG, Kamp A, Linning R, Naik S, Bakkeren G (2007) Complementation of Ustilago maydis MAPK mutants by a wheat leaf rust, Puccinia triticina homolog: potential for functional analyses of rust genes. Mol Plant Microbe Interact 20: 637–647.
[22]
Ling P, Wang MN, Chen XM, Campbell KG (2007) Construction and characterization of a full-length cDNA library for the wheat stripe rust pathogen (Puccinia striiformis f. sp. tritici). BMC Genomics 8: 145.
[23]
Chasman D, Adams RM (2001) Predicting the functional consequences of non-synonymous single nucleotide polymorphisms: structure-based assessment of amino acid variation. J Mol Biol 307: 683–706.
[24]
Koch R, van Luenen HGAM, van der Horst M, Thijssen KL, Plasterk RHA (2000) Single nucleotide polymorphisms in wild isolates of Caenorhabditis elegans. Genome Res 10: 1690–1696.
[25]
Sakaeda T, Nakamura T, Okumura K (2004) Pharmacogenetics of drug transporters and its impact on the pharmacotherapy. Curr Top Med Chem 4: 1385–1398.
[26]
Su ZG, Zhang SZ, Hou YP, Li T, Nebert DW, et al. (2002) Single-nucleotide polymorphisms in the lipoprotein lipase gene associated with coronary heart disease in Chinese. Eur J Clin Pharmacol 454: 9–18.
[27]
Forche A, Magee PT, Magee BB, May G (2004) Genome-wide single-nucleotide polymorphism map for Candida albicans. Eukaryot Cell 3: 705–714.
[28]
Mboup M, Leconte M, Gautier A, Wan AM, Chen W, et al. (2009) Evidence of genetic recombination in wheat yellow rust populations of a Chinese oversummering area. Fungal Genet Biol 46: 299–307.
[29]
Takano Y, Kikuchi T, Kubo Y, Hamer JE, Mise K, et al. (2000) The Colletotrichum lagenarium MAP kinase gene CMK1 regulates diverse aspects of fungal pathogenesis. Mol Plant Microbe Interact 13: 374–383.
[30]
Lev S, Sharon A, Hadar R, Ma H, Horwitz BA (1999) A mitogen-activated protein kinase of the corn leaf pathogen Cochliobolus heterostrophus is involved in conidiation, appressorium formation, and pathogenicity: diverse roles for mitogen-activated protein kinase homologs in foliar pathogens. Proc Natl Acad Sci U S A 96: 13542–13547.
[31]
Kinane J, Oliver RP (2003) Evidence that the appressorial development in barley powdery mildew is controlled by MAP kinase activity in conjunction with the cAMP pathway. Fungal Genet Biol 39: 94–102.
[32]
Yang J, Zhao XY, Sun J, Kang ZS, Ding SL, et al. (2010) A novel protein Com1 is required for normal conidium morphology and full virulence in Magnaporthe oryzae. Mol Plant Microbe Interact 23: 112–123.
[33]
Moldenhauer J, Moerschbacher BM, Van der Westhuizen AJ (2006) Histological investigation of stripe rust (Puccinia striiformis f. sp. tritici) development in resistant and susceptible wheat cultivars. Plant Pathol 55: 469–474.
[34]
Menotta M, Pierleoni R, Amicucci A, Sisti D, Cerasi A, et al. (2006) Characterization and complementation of a Fus3/Kss1 type MAPK from Tuber borchii, TBMK. Mol Gen Genet 276: 126–134.
[35]
Zhang YH, Qu ZP, Zheng WM, Liu B, Wang XJ, et al. (2008) Stage-specific gene expression during urediniospore germination in Puccinia striiformis f. sp. tritici. BMC Genomics 9: 203.
[36]
Hou ZM, Xue CY, Peng YL, Katan T, Kistler HC, et al. (2002) A mitogen-activated protein kinase gene (MGV1) in Fusarium graminearum is required for female fertility, heterokaryon formation, and plant infection. Mol Plant Microbe Interact 15: 1119–1127.
[37]
Wang Y, Liu WD, Hou ZM, Wang CF, Zhou XY, et al. (2011) A novel transcriptional factor important for pathogenesis and ascosporogenesis in Fusarium graminearum. Mol Plant Microbe Interact 24: 118–128.
[38]
Zhao XH, Xu JR (2007) A highly conserved MAPK-docking site in Mst7 is essential for Pmk1 activation in Magnaporthe grisea. Mol Microbio 63: 881–894.
[39]
Bruno KS, Tenjo F, Li L, Hamer JE, Xu JR (2004) Cellular localization and role of kinase activity of PMK1 in Magnaporthe grisea. Eukaryot Cell 3: 1525–1532.
[40]
Mehrabi R, Ding S, Xu JR (2008) MADS-box transcription factor Mig1 is required for infectious growth in Magnaporthe grisea. Eukaryot Cell 7: 791–799.
[41]
Sambrook J, Fritsh EF, Maniatis T (1989) Molecular cloning: A laboratory Manual. Plainview, NY: Cold Spring Harbor Laoratory Press.
[42]
Wang BT, Hu XP, Li Q, Hao BJ, Zhang B, et al. (2010) Development of race-specific SCAR markers for detection of Chinese races CYR32 and CYR33 of Puccinia striiformis f. sp. tritici. Plant Dis 94: 221–228.
[43]
Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, et al. (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31: 3497–3500.
[44]
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24: 1596–1599.
[45]
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(？Delta Delta C) method. Methods 25: 402–408.
[46]
Gong XY, Fu YP, Jiang DH, Li GQ, Yi XH, et al. (2007) L-arginine is essential for conidiation in the filamentous fungus Coniothyrium minitans. Fungal Genet Biol 44: 1368–1379.
[47]
Bluhm BH, Zhao X, Flaherty JE, Xu JR, Dunkle LD (2007) RAS2 regulates growth and pathogenesis in Fusarium graminearum. Mol Plant Microbe Interact 20: 627–636.
[48]
Proctor RH, Hohn TM, Mccormick SP (1995) Reduced virulence of Gibberella zeae caused by disruption of a trichothecene toxin biosynthetic gene. Mol Plant Microbe Interact 8: 593–601.
[49]
Gale LR, Chen LF, Hernick CA, Takamura K, Kistler HC (2002) Population analysis of Fusarium graminearum from wheat fields in eastern China. Phytopathology 92: 1315–1322.
[50]
Seong K, Li L, Hou ZM, Tracy M, Kistler HC, et al. (2006) Cryptic promoter activity in the coding region of the HMG-CoA rediactase gene in Fusarium graminearum. Fungal Genet Biol 43: 34–41.
[51]
Ding SL, Liu WD, Iliuk A, Ribot C, Vallet J, et al. (2010) The Tig1 histone deacetylase complex regulates infectious growth in the rice blast fungus Magnaporthe oryzae. Plant Cell 22: 2495–2508.
[52]
Xu JR, Urban M, Sweigard JA, Hamer JE (1997) The CPKA gene of Magnaporthe grisea is essential for appressorial penetration. Mol Plant Microbe Interact 10: 187–194.
[53]
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 U S A 95: 12713–12718.
