Characterization of the Largest Effector Gene Cluster of

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In this study, we provide the first step to the functional analysis of the largest gene cluster in the Ustilago maydis genome encoding 24 secreted effectors. While the deletion of the entire cluster dramatically affected tumor formation and abolished anthocyanin induction, only one of the genes had a large contribution to tumor formation, while another effector gene was primarily responsible for the anthocyanin induction. Unexpectedly, the cluster mutant could still colonize plants and complete the life cycle, i.e. behaves like an endophyte. Despite only small contributions to tumor formation, individual effector mutants caused distinct plant responses, suggesting that they affect discrete plant processes. On these grounds we are proposing to use plant responses as a general readout to assess and compare effector gene function.

Vyšlo v časopise: Characterization of the Largest Effector Gene Cluster of. PLoS Pathog 10(7): e32767. doi:10.1371/journal.ppat.1003866
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003866

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Zdroje

1. BrefortT, DoehlemannG, Mendoza-MendozaA, ReissmannS, DjameiA, et al. (2009) Ustilago maydis as a pathogen. Annu Rev Phytopathol 47: 423–445.

2. VollmeisterE, SchipperK, BaumannS, HaagC, PohlmannT, et al. (2012) Fungal development of the plant pathogen Ustilago maydis. FEMS Microbiol Rev 36: 59–77.

3. Mendoza-MendozaA, BerndtP, DjameiA, WeiseC, LinneU, et al. (2009) Physical-chemical plant-derived signals induce differentiation in Ustilago maydis. Mol Microbiol 71: 895–911.

4. SchirawskiJ, BohnertHU, SteinbergG, SnetselaarK, AdamikowaL, et al. (2005) Endoplasmic reticulum glucosidase II is required for pathogenicity of Ustilago maydis. Plant Cell 17: 3532–3543.

5. BauerR, OberwinklerF, VankyK (1997) Ultrastructural markers and systematics in smut fungi and allied taxa. Can J Bot 75: 1273–1314.

6. DoehlemannG, WahlR, VranesM, de VriesRP, KaemperJ, et al. (2008) Establishment of compatibility in the Ustilago maydis/maize pathosystem. J Plant Physiol 165: 29–40.

7. SchirawskiJ, MannhauptG, MuenchK, BrefortT, SchipperK, et al. (2010) Pathogenicity determinants in smut fungi revealed by genome comparison. Science 330: 1546–1548.

8. LaurieJD, AliS, LinningR, MannhauptG, WongP, et al. (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.

9. DoehlemannG, WahlR, HorstRJ, VollLM, UsadelB, et al. (2008) Reprogramming a maize plant: transcriptional and metabolic changes induced by the fungal biotroph Ustilago maydis. Plant J 56: 181–195.

10. van der LindeK, HemetsbergerC, KastnerC, KaschaniF, van der HoornRAL, et al. (2012) A maize cystatin suppresses host immunity by inhibiting apoplastic cysteine proteases. Plant Cell 24: 1285–1300.

11. WenzlerH, MeinsF (1987) Persistent changes in the proliferative capacity of maize leaf tissues induced by Ustilago infection. Physiol Mol Plant Pathol 30: 309–319.

12. KaemperJ, KahmannR, BoelkerM, MaL-J, BrefortT, et al. (2006) Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis. Nature 444: 97–101.

13. MuellerO, KahmannR, AguilarG, Trejo-AguilarB, WuA, et al. (2008) The secretome of the maize pathogen Ustilago maydis. Fungal Genet Biol 45: S63–S70.

14. SkibbeDS, DoehlemannG, FernandesJ, WalbotV (2010) Maize tumors caused by Ustilago maydis require organ-specific genes in host and pathogen. Science 328: 89–92.

15. DoehlemannG, van der LindeK, AssmannD, SchwammbachD, HofA, et al. (2009) Pep1, a secreted effector protein of Ustilago maydis, is required for successful invasion of plant cells. PLoS pathogens 5: e1000290–e1000290.

16. HemetsbergerC, HerrbergerC, ZechmannB, HillmerM, DoehlemannG (2012) The Ustilago maydis effector Pep1 suppresses plant immunity by inhibition of host peroxidase activity. PLoS Pathog 8: e1002684 doi:10.1371/journal.ppat.1002684

17. DoehlemannG, ReissmannS, AssmannD, FleckensteinM, KahmannR (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.

18. MuellerAN, ZiemannS, TreitschkeS, AssmannD, DoehlemannG (2013) Compatibility in the Ustilago maydis-maize interaction requires inhibition of host cysteine proteases by the fungal effector Pit2. PLoS Pathog 9: e1003177 doi:10.1371/journal.ppat.1003177

19. DjameiA, SchipperK, RabeF, GhoshA, VinconV, et al. (2011) Metabolic priming by a secreted fungal effector. Nature 478: 395–398.

20. WahlR, ZahiriA, KaemperJ (2010) The Ustilago maydis b mating type locus controls hyphal proliferation and expression of secreted virulence factors in planta. Mol Microbiol 75: 208–220.

21. van der LindeK, KastnerC, KumlehnJ, KahmannR, DoehlemannG (2011) Systemic virus-induced gene silencing allows functional characterization of maize genes during biotrophic interaction with Ustilago maydis. New Phytol 189: 471–483.

22. LiJ, BraderG, HeleniusE, KariolaT, PalvaET (2012) Biotin deficiency causes spontaneous cell death and activation of defense signaling. Plant J 70: 315–326.

23. Chalker-ScottL (1999) Environmental significance of anthocyanins in plant stress responses. Photochem Photobiol 70: 1–9.

24. SteynWJ, WandSJE, HolcroftDM, JacobsG (2002) Anthocyanins in vegetative tissues: a proposed unified function in photoprotection. New Phytol 155: 349–361.

25. VignaisPV (2002) The superoxide-generating NADPH oxidase: structural aspects and activation mechanism. Cell Mol Life Sci 59: 1428–1459.

26. SegondD, DellagiA, LanquarV, RigaultM, PatritO, et al. (2009) NRAMP genes function in Arabidopsis thaliana resistance to Erwinia chrysanthemi infection. Plant J 58: 195–207.

27. LiuG, GreenshieldsDL, SammynaikenR, HirjiRN, SelvarajG, et al. (2007) Targeted alterations in iron homeostasis underlie plant defense responses. J Cell Sci 120: 596–605.

28. JooY-C, OhD-K (2012) Lipoxygenases: Potential starting biocatalysts for the synthesis of signaling compounds. Biotechnol Adv 30: 1524–1532.

29. FengY, XueQ (2006) The serine carboxypeptidase like gene family of rice (Oryza sativa L. ssp japonica). Funct Integr Genomic 6: 14–24.

30. LiuH, WangX, ZhangH, YangY, GeX, et al. (2008) A rice serine carboxypeptidase-like gene OsBISCPL1 is involved in regulation of defense responses against biotic and oxidative stress. Gene 420: 57–65.

31. AliS, LaurieJD, LinningR, Cervantes-ChávezJA, GaudetD, et al. (2014) An immunity-triggering effector from the barley smut fungus Ustilago hordei resides in an Ustilaginaceae-specific cluster bearing signs of transposable element-assisted evolution. PLoS Pathog 10: e1004223.

32. StokesME, ChattopadhyayA, WilkinsO, NambaraE, CampbellMM (2013) Interplay between sucrose and folate modulates auxin signaling in arabidopsis. Plant Physiol 162: 1552–1565.

33. MillerG, ShulaevV, MittlerR (2008) Reactive oxygen signaling and abiotic stress. Physiol Plantarum 133: 481–489.

34. MassonneauA, Houba-HérinN, PetheC, MadzakC, FalqueM, et al. (2004) Maize cytokinin oxidase genes: differential expression and cloning of two new cDNAs. J Exp Bot 55: 2549–2557.

35. RafiqiM, EllisJG, LudowiciVA, HardhamAR, DoddsPN (2012) Challenges and progress towards understanding the role of effectors in plant-fungal interactions. Curr Opin Plant Biol 15: 477–482.

36. WinJ, KrasilevaKV, KamounS, ShirasuK, StaskawiczBJ, et al. (2012) Sequence divergent RXLR effectors share a structural fold conserved across plant pathogenic oomycete species. PLoS Pathog 8: e1002400 doi:10.1371/journal.ppat.1002400

37. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

38. LoubradouG, BrachmannA, FeldbruggeM, KahmannR (2001) A homologue of the transcriptional repressor Ssn6p antagonizes cAMP signalling in Ustilago maydis. Mol Microbiol 40: 719–730.

39. HoffmanCS, WinstonF (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of E. coli. Gene 57: 267–272.

40. LivakKJ, SchmittgenTD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2((−ΔΔC(T)) Method. Methods 25: 402–408.

41. EisenhartC (1947) The assumptions underlying the analysis of variance. Biometrics 3: 1–21.

42. BrachmannA, SchirawskiJ, MullerP, KahmannR (2003) An unusual MAP kinase is required for efficient penetration of the plant surface by Ustilago maydis. EMBO J 22: 2199–2210.

43. ZhengQ, WangX-J (2008) GOEAST: a web-based software toolkit for Gene Ontology enrichment analysis. Nucleic Acids Res 36: W358–W363.