Abstract
Peroxisomes are indispensable organelles that play critical roles in various biological processes in eukaryotic cells. PEX4, one of the peroxins, is the ubiquitin-conjugating enzyme. To functionally characterize roles of FgPEX4 in the phytopathogenic fungus, Fusarium graminearum, we constructed a deletion mutant of FgPEX4 (ΔPEX4) through homologous recombination. ΔPEX4 displayed reduced mycelial growth, conidiation, and the production of perithecia. ΔPEX4 was defective in pathogenicity and production of the mycotoxin deoxynivalenol (DON). In addition, FgPEX4 was involved in cell wall integrity, lipid droplet accumulation, and the elimination of reactive oxygen species. Western blot analysis revealed reduced phosphorylation of Mgv1 in the ∆PEX4 mutant. Importantly, proteomics analysis indicated that protein expression levels related to protein biosynthesis, fatty acid metabolism, cell wall synthesis, and oxidation–reduction reactions were downregulated in ΔPEX4 compared with the wild type. Taken together, these results demonstrate that FgPEX4 is important for development, pathogenicity, and cell wall integrity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bai G, Shaner G (2004) Management and resistance in wheat and barley to Fusarium head blight. Annu Rev Phytopathol 42:135–161. https://doi.org/10.1146/annurev.phyto.42.040803.140340
Beller M, Thiel K, Thul PJ, Jäckle H (2010) Lipid droplets: a dynamic organelle moves into focus. FEBS Lett 584:2176–2182. https://doi.org/10.1016/j.febslet.2010.03.022
Binns D, Januszewski T, Chen Y, Hill J, Markin VS, Zhao Y (2006) An intimate collaboration between peroxisomes and lipid bodies. J Cell Biol 173:719–731. https://doi.org/10.1083/jcb200511125
Bowden RL, Leslie JF (1999) Sexual recombination in Gibberella zeae. Phytopathology 89:182–188. https://doi.org/10.1094/PHYTO.1999.89.2.182
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. https://doi.org/10.1128/EC.3.6.1525-1532.2004
Carballeira NM (2008) New advances in fatty acids as antimalarial, antimycobacterial and antifungal agents. Prog Lipid Res 47:50–61. https://doi.org/10.1016/j.plipres.2007.10.002
Catlett NL, Lee BN, Yoder OC, Turgeon BG (2003) Split-marker recombination for efficient targeted deletion of fungal genes. Fungal Genet Newsl 50:9–11. https://doi.org/10.4148/1941-4765.1150
Chayakulkeeree M, Sorrell TC, Siafakas AR, Wilson CF, Pantarat N, Gerik KJ, Boadle RA, Djordjevic JT (2008) Role and mechanism of phosphatidylinositol-specific phospholipase C in survival and virulence of Cryptococcus neoformans. Mol Microbiol 69:809–826. https://doi.org/10.1111/j.1365-2958.2008.06310.x
Chen XL, Wang Z, Liu C (2016) Roles of peroxisomes in the rice blast fungus. Biomed Res Int ID 9343417. https://doi.org/10.1155/2016/9343417
Chen Y, Zheng S, Ju Z, Zhang C, Tang G, Wang J, Wen Z, Chen W, Ma Z (2018) Contribution of peroxisomal docking machinery to mycotoxin biosynthesis, pathogenicity and pexophagy in the plant pathogenic fungus Fusarium graminearum. Environ Microbiol 20:3224–3245. https://doi.org/10.1111/1462-2920.14291
Dammai V, Subramani S (2001) The human peroxisomal targeting signal receptor, Pex5p, is translocated into the peroxisomal matrix and recycled to the cytosol. Cell 105:187–196. https://doi.org/10.1016/S0092-8674(01)00310-5
Dean R, Jan AL, Pretorius KANV, Hammond ZA, Pietro KE, Spanu AD, Rudd PD, Dickman JJ, Kahmann M, Ellis R, Foster J GD (2012) The Top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol 13:414–430. https://doi.org/10.1111/J.1364-3703.2011.00783.X
Deng SZ, Gu ZK, Yang NY, Li L, Yue XF, Que YW, Sun GC, Wang ZY, Wang JY (2016) Identification and characterization of the peroxin 1 gene MoPEX1 required for infection-related morphogenesis and pathogenicity in Magnaporthe oryzae. Sci Rep 6:36292. https://doi.org/10.1038/srep36292
Der Klei IJV, Hilbrands RE, Kiel JAKW, Rasmussen SW, Cregg JM, Veenhuis M (1998) The ubiquitin-conjugating enzyme Pex4p of Hansenula polymorpha is required for efficient functioning of the PTS1 import machinery. EMBO J 17:3608–3618. https://doi.org/10.1093/emboj/17.13.3608
Desjardins AE, Hohn TM, Mccormick SP (1993) Trichothecene biosynthesis in Fusarium species: chemistry, genetics, and significance. Microbiol Rev 57:595–604
Ding MY, Li J, Fan X, He F, Yu XY, Liang YC, Yu JF (2018) Aquaporin1 regulates development, secondary metabolism and stress responses in Fusarium graminearum. Curr Genet 64:1057–1069. https://doi.org/10.1007/s00294-018-0818-8
Distel B, Erdmann R, Gould AJ, Blobel G, Crane DI, Cregg JM, Dodt G, Fujiki Y, Goodman JM, Just WW, Kiel JAKW, Kunau WH, Lazarow PB, Mannaerts GP, Moser HW, Osumi T (1996) A unified nomenclature for peroxisome biogenesis factors. J Cell Biol 135:1–3
Farese RV, Walther TC (2009) Lipid droplets finally get a little R-E-S-P-E-C-T. Cell 139:855–860. https://doi.org/10.1016/j.cell.2009.11.005
Faust PL, Banka D, Siriratsivawong R, Ng VG, Wikander TM (2005) Peroxisome biogenesis disorders: the role of peroxisomes and metabolic dysfunction in developing brain. J Inherit Metab Dis 28:369–383. https://doi.org/10.1007/s10545-005-7059-y
Gale LR, Ward TJ, Balmas V, Kistler HC (2007) Population subdivision of Fusarium graminearum sensu stricto in the upper midwestern united states. Phytopathology 97:1434–1439. https://doi.org/10.1094/PHYTO-97-11-1434
Goh J, Jeon J, Kim KS, Park J, Park SY, Lee YH (2011) The PEX7-mediated peroxisomal import system is required for fungal development and pathogenicity in Magnaporthe oryzae. Plos ONE 6:e28220. https://doi.org/10.1371/journal.pone.0028220
Goswami RS, Kistler HC (2010) Heading for disaster: Fusarium graminearum on cereal crops. Mol Plant Pathol 5:515–525. https://doi.org/10.1111/j.1364-3703.2004.00252.x
Gould SJ, Mccollum D, Spong AP, Heyman JA, Subramani S (2010) Development of the yeast Pichia pastoris as a model organism for a genetic and molecular analysis of peroxisome assembly. Yeast 8:613–628. https://doi.org/10.1002/yea.320080805
Hamel LP, Nicole MC, Duplessis S, Ellis BE (2012) Mitogen-activated protein kinase signaling in plant-interacting fungi: distinct messages from conserved messengers. Plant Cell 24:1327–1351
Heinisch JJ, Lorberg A, Schmitz HP, Jacoby JJ (1999) The protein kinase C-mediated MAP kinase pathway involved in the maintenance of cellular integrity in Saccharomyces cerevisiae. Mol Microbiol 32:671–680
Hiltunen JK, Mursula AM, Rottensteiner H, Wierenga RK, Kastaniotis AJ, Gurvitz A (2003) The biochemistry of peroxisomal β-oxidation in the yeast Saccharomyces cerevisiae. FEMS Microbiol Rev 27:35–64. https://doi.org/10.1016/S0168-6445(03)00017-2
Hou Z, Xue C, Peng Y, Katan T, Kistler HC, Xu JR (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. https://doi.org/10.1094/MPMI.2002.15.11.1119
Hu J, Baker A, Bartel B, Linka N, Mullen RT, Reumann S, Zolman BK (2012) Plant peroxisomes: biogenesis and function. Plant Cell 24:2279–2303. https://doi.org/10.1105/tpc.112.096586
Imazaki A, Tanaka A, Harimoto Y, Yamamoto M, Akimitsu K, Park P, Tsuge T (2010) Contribution of peroxisomes to secondary metabolism and pathogenicity in the fungal plant pathogen Alternaria alternata. Eukaryot Cell 9:682–694. https://doi.org/10.1128/EC.00369-09
Jenczmionka NJ, Maier FJ, Lösch AP, Schäfer 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. https://doi.org/10.1007/s00294-003-0379-2
Jiang C, Zhang X, Liu HQ, Xu JR (2018) Mitogen-activated protein kinase signaling in plant pathogenic fungi. Plos Pathog 14(3):e1006875
Leslie JF, Summerell BA (2007) The Fusarium laboratory manual. Willy, Hoboken
Li L, Wang J, Zhang Z, Wang Y, Liu M, Jiang H, Chai R, Mao XQ, Qiu HP, Liu FQ, Sun GC (2014) MoPex19, which is essential for maintenance of peroxisomal structure and woronin bodies, is required for metabolism and development in the rice blast fungus. Plos ONE 9:e85252. https://doi.org/10.1371/journal.pone.0085252
Li L, Wang J, Chen H, Chai R, Zhang Z, Mao X, Qiu HP, Jiang H, Wang Y, Sun GC (2016) Pex14/17, a filamentous fungus specific peroxin, is required for the import of peroxisomal matrix proteins and full virulence of Magnaporthe oryzae. Mol Plant Pathol 18:1238–1252. https://doi.org/10.1111/mpp.12487
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT Method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
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. https://doi.org/10.1007/s00294-009-0259-5
Matsuzono Y, Kinoshita N, Tamura S, Shimozawa N, Hamasaki M, Ghaedi K, Wanders RJA, Suzuki Y, Kondo N, Fujiki Y (1999) Human PEX19: cDNA cloning by functional complementation, mutation analysis in a patient with Zellweger syndrome, and potential role in peroxisomal membrane assembly. Proc Natl Acad Sci USA 96:2116–2121. https://doi.org/10.1073/pnas.96.5.2116
McMullen M, Jones R, Gallenberg D (2007) Scab of wheat and barley: a re-emerging disease of devastating impact. Plant Dis 81:1340–1348
Meijer WH, Ij van der Klei IJ, Veenhuis M, Kiel JAKW (2007) ATG genes involved in non-selective autophagy are conserved from yeast to man, but the selective Cvt and pexophagy pathways also require organism-specific genes. Autophagy 3:106–116. https://doi.org/10.4161/auto.3595
Min K, Son H, Lee J, Choi GJ, Kim JC, Lee YW (2012) Peroxisome function is required for virulence and survival of Fusarium graminearum. Mol Plant Microbe Interact 25:1617–1627. https://doi.org/10.1094/MPMI-06-12-0149-R
Mirocha CJ, Kolaczkowski E, Xie W, Yu H, Jelen H (1998) Analysis of deoxynivalenol and its derivatives (batch and single kernel) using gas chromatography/mass spectrometry. J Agric Food Chem 46:1414–1418
Pestka JJ, Smolinski AT (2005) Deoxynivalenol: toxicology and potential effects on humans. J Toxicol Environ Heal B 8:39–69. https://doi.org/10.1080/10937400590889458
Petter M, Johan S, Wagner EGH (2003) Characterization of Phia, a gene essential for phialide development in Aspergillus Nidulans. Fungal Genet Biol 40:234–241. https://doi.org/10.1016/S1087-1845(03)00108-7
Pieuchot L, Jedd G (2012) Peroxisome assembly and functional diversity in eukaryotic microorganisms. Annu Rev Microbiol 66:237–263. https://doi.org/10.1146/annurev-micro-092611-150126
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
Qin J, Wang G, Jiang C, Xu JR, Wang C (2015) Fgk3 glycogen synthase kinase is important for development, pathogenesis, and stress responses in Fusarium graminearum. Sci Rep 5:8504. https://doi.org/10.1038/srep08504
Ramospamplona M, Naqvi NI (2006) Host invasion during rice-blast disease requires carnitine-dependent transport of peroxisomal acetyl-CoA. Mol Microbiol 61:61–75. https://doi.org/10.1111/j.1365-2958.2006.05194.x
Seong K, Hou Z, Tracy M, Kistler HC, Xu JR (2005) Random insertional mutagenesis identifies genes associated with virulence in the wheat scab fungus Fusarium graminearum. Phytopathology 95:744–750. https://doi.org/10.1094/PHYTO-95-0744
Titorenko VI, Rachubinski RA (2001) The life cycle of the peroxisome. Nat Rev Mol Cell Bio 2:357–368. https://doi.org/10.1038/35073063
Vizeacoumar FJ, Torres-Guzman JC, Tam YYC, Aitchison JD, Rachubinski RA (2003) YHR150w and YDR479c encode peroxisomal integral membrane proteins involved in the regulation of peroxisome number, size, and distribution in Saccharomyces cerevisiae. J Cell Biol 161:321–332. https://doi.org/10.1083/jcb.200210130
Wanders RJ (2004) Peroxisomes, lipid metabolism, and peroxisomal disorders. Mol Genet Metab 83:16–27. https://doi.org/10.1016/j.bbamcr.2018.01.003
Wanders RJ, Waterham HR (2010) Peroxisomal disorders I: biochemistry and genetics of peroxisome biogenesis disorders. Clin Genet 67:107–133. https://doi.org/10.1111/j.1399-0004.2004.00329.x
Wang ZY, Soanes DM, Kershaw MJ, Talbot NJ (2007) Functional analysis of lipid metabolism in Magnaporthe grisea reveals a requirement for peroxisomal fatty acid beta-oxidation during appressorium-mediated plant infection. Mol Plant Microbe Interact 20:475–491. https://doi.org/10.1094/MPMI-20-5-0475
Wiebel FF, Kunau WH (1992) The Pas2 protein essential for peroxisome biogenesis is related to ubiquitin-conjugating enzymes. Nature 359:73–79
Williams C, Marlene B, Santosh P, Will AS, Ben D, Matthias W (2012) Insights into ubiquitin-conjugating enzyme/co-activator interactions from the structure of the Pex4p: Pex22p complex. EMBO J 31:391–402. https://doi.org/10.1038/emboj.2011.411
Xu JR, Staiger CJ, Hamer JE (1998) Inactivation of the mitogen activated protein kinase MPS1 in the rice blast fungus prevents penetration of host cells but allows activation of plant defense responses. Proc Natl Acad Sci USA 95:12713–12718
Zhang F, Geng LP, Huang LH, Deng JL, Opemipo EF, Guangshan Y, Wang SH (2018) Contribution of peroxisomal protein importer AflPex5 to development and pathogenesis in the fungus Aspergillus flavus. Curr Genet 64:1335–1348. https://doi.org/10.1007/s00294-018-0851-7
Zhou X, Li G, Xu JR (2011) Efficient approaches for generating GFP fusion and epitope-tagging constructs in filamentous fungi. Methods Mol Biol 722:199–212. https://doi.org/10.1007/978-1-61779-040-9-15
Acknowledgements
We thank Larry Dunkle (Emeritus Professor, Purdue University, USA) for improving this manuscript. This work was supported by Agro-Industry R & D Special fund of China (2017YFD0201705): integration and demonstration of Chemical Fertilizer reduction Technology for Winter Wheat in East Shandong, the Wheat Innovation Team of Shandong Province Modern Agricultural Industry Technology System (SDAIT-01-09), and Funds of Shandong “Double Tops” Program (SYL2017XTTD11).
Author information
Authors and Affiliations
Corresponding authors
Additional information
Communicated by M. Kupiec.
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Zhang, L., Wang, L., Liang, Y. et al. FgPEX4 is involved in development, pathogenicity, and cell wall integrity in Fusarium graminearum. Curr Genet 65, 747–758 (2019). https://doi.org/10.1007/s00294-018-0925-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00294-018-0925-6