Australasian Plant Pathology

, Volume 43, Issue 5, pp 535–546 | Cite as

Fungal growth, proteinaceous toxins and virulence of Pyrenophora teres f. teres on barley

Article

Abstract

Pyrenophora teres f. teres (Ptt) causes net form net blotch (NFNB), an important disease of barley, but isolates of Ptt vary in their ability to cause symptoms on susceptible cultivars. Ptt isolates with different virulence were used to compare conidial germination and fungal growth on the barley cultivar ‘Sloop’. Whether proteinaceous toxins from culture filtrates of the six isolates or different fractions and sub-fractions of those filtrates induced different symptoms was also investigated. Greater conidial germination and appressorial formation was observed during infection by more virulent isolates but hyphal length was variable. Even though the six isolates varied in virulence from low to high, the proteinaceous toxins extracted from culture filtrates of all isolates were able to induce necrosis when injected into barley leaves. Ptt isolates therefore appear genetically able to produce proteinaceous toxins and the difference in virulence between isolates may reflect the growth of the fungus and the capacity for toxins to be delivered to the plant tissue. Proteins identified in the biologically active fractions included glycoside hydrolase, cysteine hydrolase, CFEM (common in fungal extracellular membrane) domain-containing protein, lactonase and peptidase. These have been previously suggested to have roles in plant cell wall degradation, fungal growth and/or host-pathogen interactions.

Keywords

Barley net blotch Net form net blotch Virulence Proteinaceous toxins Fungal growth 

Notes

Acknowledgments

We thank Dr Hugh Wallwork, SARDI, for providing Ptt isolates and the Grains Research and Development Corporation (GRDC) for supporting this research. IAI was supported by a scholarship from the Iraqi Ministry of Higher Education and Scientific Research.

Supplementary material

13313_2014_295_MOESM1_ESM.docx (576 kb)
ESM 1 (DOCX 575 kb)

References

  1. Able AJ (2003) Role of reactive oxygen species in the response of barley to necrotrophic pathogens. Protoplasma 221:137–143PubMedCrossRefGoogle Scholar
  2. Able AJ, Guest DI, Sutherland MW (1998) Use of a new tetrazolium-based assay to study the production of superoxide radicals by tobacco cell cultures challenged with avirulent zoospores of Phytophthora parasitica var nicotianae. Plant Physiol 117:491–499PubMedCentralPubMedCrossRefGoogle Scholar
  3. Afanasenko OS, Mironenko NV, Filatova OA, Serenius M (2007) Structure of Pyrenophora teres f. teres populations from Leningrad region and Finland by virulence. Mikol Fitopatol 41:261–268Google Scholar
  4. Aizat WM, Able JA, Stangoulis JCR, Able AJ (2013) Proteomic analysis during capsicum ripening reveals differential expression of ACC oxidase isoform 4 and other candidates. Funct Plant Biol 40:1115–1128CrossRefGoogle Scholar
  5. Anguelova-Merhar VS, Van Der Westhuizen AJ, Pretorius ZA (2001) β-1,3-glucanase and chitinase activities and the resistance response of wheat to leaf rust. J Phytopathol 149:381–384CrossRefGoogle Scholar
  6. ApelBirkhold PC, Walton JD (1996) Cloning, disruption, and expression of two endo-beta 1,4-xylanase genes, XYL2 and XYL3, from Cochliobolus carbonum. Appl Environ Microbiol 62:4129–4135Google Scholar
  7. Beliën T, Van Campenhout S, Robben J, Volckaert G (2006) Microbial endoxylanases: effective weapons to breach the plant cell-wall barrier or, rather, triggers of plant defense systems? Mol Plant-Microbe Interact 19:1072–1081PubMedCrossRefGoogle Scholar
  8. Bendtsen JD, Jensen LJ, Blom N, von Heijne G, Brunak S (2004) Feature-based prediction of non-classical and leaderless protein secretion. Protein Eng Des Sel 17:349–356PubMedCrossRefGoogle Scholar
  9. Bent AF, Mackey D (2007) Elicitors, effectors, and R genes: the new paradigm and a lifetime supply of questions. Annu Rev Phytopathol 45:399–436PubMedCrossRefGoogle Scholar
  10. Boonvitthya N, Tanapong P, Kanngan P, Burapatana V, Chulalaksananukul W (2012) Cloning and expression of the Aspergillus oryzae glucan 1,3-beta-glucosidase A (exgA) in Pichia pastoris. Biotechnol Lett 34:1937–1943PubMedCrossRefGoogle Scholar
  11. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  12. Brown MP, Steffenson BJ, Webster RK (1993) Host-range of Pyrenophora teres f. teres isolates from California. Plant Dis 77:942–947CrossRefGoogle Scholar
  13. DeZwaan TM, Carroll AM, Valent B, Sweigard JA (1999) Magnaporthe grisea Pth11p is a novel plasma membrane protein that mediates appressorium differentiation in response to inductive substrate cues. Plant Cell 11:2013–2030PubMedCentralPubMedCrossRefGoogle Scholar
  14. Dushnicky LG, Ballance GM, Sumner MJ, MacGregor AW (1998) The role of lignification as a resistance mechanism in wheat to a toxin-producing isolate of Pyrenophora tritici repentis. Can J Plant Pathol 20:35–47CrossRefGoogle Scholar
  15. El-Bebany AF, Rampitsch C, Daayf F (2010) Proteomic analysis of the phytopathogenic soilborne fungus Verticillium dahliae reveals differential protein expression in isolates that differ in aggressiveness. Proteomics 10:289–303PubMedCrossRefGoogle Scholar
  16. Ellouze OE, Loukil S, Marzouki MN (2011) Cloning and molecular characterization of a new fungal xylanase gene from Sclerotinia sclerotiorum S2. BMB Rep 44:653–658PubMedCrossRefGoogle Scholar
  17. Fernando THPS, Jayasinghe CK, Wijesundera RLC (2000) Factors affecting spore production, germination and viability of Colletotrichum acutatum isolates from Hevea brasiliensis. Mycol Res 104:681–685CrossRefGoogle Scholar
  18. Fiegen M, Knogge W (2002) Amino acid alterations in isoforms of the effector protein NIP1 from Rhynchosporium secalis have similar effects on its avirulence- and virulence-associated activities on barley. Physiol Mol Plant Pathol 61:299–302CrossRefGoogle Scholar
  19. Friesen TL, Stukenbrock EH et al (2006) Emergence of a new disease as a result of interspecific virulence gene transfer. Nat Genet 38:953–956PubMedCrossRefGoogle Scholar
  20. Gowda M, Venu RC et al (2006) Deep and comparative analysis of the mycelium and appressorium transcriptomes of Magnaporthe grisea using MPSS, RL-SAGE, and oligoarray methods. BMC Genomics 7:1–15CrossRefGoogle Scholar
  21. Hartl L, Gastebois A, Aimanianda V, Latgé JP (2011) Characterization of the GPI-anchored endo β-1,3-glucanase Eng2 of Aspergillus fumigatus. Fungal Genet Biol 48:185–191PubMedCentralPubMedCrossRefGoogle Scholar
  22. Jalli M (2011) Sexual reproduction and soil tillage effects on virulence of Pyrenophora teres in Finland. Ann Appl Biol 158:95–105CrossRefGoogle Scholar
  23. Kennedy EJ, Pillus L, Ghosh G (2005) Pho5p and newly identified nucleotide pyrophosphatases/ phosphodiesterases regulate extracellular nucleotide phosphate metabolism in Saccharomyces cerevisiae. Eukaryot Cell 4:1892–1901PubMedCentralPubMedCrossRefGoogle Scholar
  24. Keon JPR, Hargreaves JA (1983) A cytological study of the net blotch disease of barley caused by Pyrenophora teres. Physiol Plant Pathol 22:321–329CrossRefGoogle Scholar
  25. Kokkelink L, Minz A, Al-Masri M, Giesbert S, Barakat R, Sharon A, Tudzynski P (2011) The small GTPase BcCdc42 affects nuclear division, germination and virulence of the gray mold fungus Botrytis cinerea. Fungal Genet Biol 48:1012–1019PubMedCrossRefGoogle Scholar
  26. Kopetz VA, Penno MAS, Hoffmann P, Wilson DP, Beltrame JF (2012) Potential mechanisms of the acute coronary syndrome presentation in patients with the coronary slow flow phenomenon - Insight from a plasma proteomic approach. Int J Acarol 156:84–91Google Scholar
  27. Kulkarni RD, Kelkar HS, Dean RA (2003) An eight-cysteine-containing CFEM domain unique to a group of fungal membrane proteins. Trends Biochem Sci 28:118–121PubMedCrossRefGoogle Scholar
  28. Lehmensiek A, Bester-van der Merwe AE, Sutherland MW, Platz G, Kriel WM, Potgieter GF, Prins R (2010) Population structure of South African and Australian Pyrenophora teres isolates. Plant Pathol 59:504–515CrossRefGoogle Scholar
  29. Lightfoot DJ, Able AJ (2010) Growth of Pyrenophora teres in planta during barley net blotch disease. Australas Plant Pathol 39:499–507CrossRefGoogle Scholar
  30. Liu Z, Ellwood SR, Oliver RP, Friesen TL (2011) Pyrenophora teres: profile of an increasingly damaging barley pathogen. Mol Plant Pathol 12:1–19PubMedCrossRefGoogle Scholar
  31. Liu ZH, Zhong S, Stasko AK, Edwards MC, Friesen TL (2012) Virulence profile and genetic structure of a North Dakota population of Pyrenophora teres f. teres, the causal agent of net form net blotch of barley. Phytopathology 102:539–546PubMedCrossRefGoogle Scholar
  32. Martin-Cuadrado AB, Duenas E, Sipiczki M, de Aldana CRV, del Rey F (2003) The endo-beta-1,3-glucanase eng1p is required for dissolution of the primary septum during cell separation in Schizosaccharomyces pombe. J Cell Sci 116:1689–1698PubMedCrossRefGoogle Scholar
  33. Mouyna I, Fontaine T, Vai M, Monod M, Fonzi WA, Diaquin M, Popolo L, Hartland RP, Latgé J-P (2000) Glycosylphosphatidylinositol-anchored Glucanosyltransferases play an active role in the biosynthesis of the fungal cell wall. J Biol Chem 275:14882–14889PubMedCrossRefGoogle Scholar
  34. Murray GM, Brennan JP (2010) Estimating disease losses to the Australian barley industry. Australas Plant Pathol 39:85–96CrossRefGoogle Scholar
  35. Nguyen QB, Itoh K, Van Vu B, Tosa Y, Nakayashiki H (2011) Simultaneous silencing of endo-β-1,4 xylanase genes reveals their roles in the virulence of Magnaporthe oryzae. Mol Microbiol 81:1008–1019PubMedCrossRefGoogle Scholar
  36. Okinaka Y, Mimori K, Takeo K, Kitamura S, Takeuchi Y, Yamaoka N, Yoshikawa M (1995) A structural model for the mechanisms of elicitor release from fungal cell-walls by plant beta-1,3-endoglucanase. Plant Physiol 109:839–845PubMedCentralPubMedCrossRefGoogle Scholar
  37. Rau D, Brown AHD, Brubaker CL, Attene G, Balmas V, Saba E, Papa R (2003) Population genetic structure of Pyrenophora teres Drechs. the causal agent of net blotch in Sardinian landraces of barley (Hordeum vulgare L.). Theor Appl Genet 106:947–959PubMedGoogle Scholar
  38. Sarpeleh A, Wallwork H, Catcheside DEA, Tate ME, Able AJ (2007) Proteinaceous metabolites from Pyrenophora teres contribute to symptom development of barley net blotch. Phytopathology 97:907–915PubMedCrossRefGoogle Scholar
  39. Sarpeleh A, Wallwork H, Tate ME, Catcheside DEA, Able AJ (2008) Initial characterisation of phytotoxic proteins isolated from Pyrenophora teres. Physiol Mol Plant Pathol 72:73–79CrossRefGoogle Scholar
  40. Serenius M, Manninen O, Wallwork H, Williams K (2007) Genetic differentiation in Pyrenophora teres populations measured with AFLP markers. Mycol Res 111:213–223PubMedCrossRefGoogle Scholar
  41. Soanes DM, Alam I et al (2008) Comparative genome analysis of filamentous fungi reveals gene family expansions associated with fungal pathogenesis. PLoS ONE 3:e2300PubMedCentralPubMedCrossRefGoogle Scholar
  42. Stanford DR, Whitney ML, Hurto RL, Eisaman DM, Shen WC, Hopper AK (2004) Division of labor among the yeast sol proteins implicated in tRNA nuclear export and carbohydrate metabolism. Genetics 168:117–127PubMedCentralPubMedCrossRefGoogle Scholar
  43. Stleger RJ, Bidochka MJ, Roberts DW (1994) Characterization of a novel carboxypeptidase produced by the entomopathogenic fungus Metarhizium anisopliae. Arch Biochem Biophys 314:392–398CrossRefGoogle Scholar
  44. Tan K, Oliver RP, Solomon PS, Moffat CS (2010) Proteinaceous necrotrophic effectors in fungal virulence. Funct Plant Biol 37:907–912CrossRefGoogle Scholar
  45. Tekauz A (1985) A numerical scale to classify reactions of barley to Pyrenophora teres. Can J Plant Pathol 7:181–183CrossRefGoogle Scholar
  46. Vancaeseele L, Grumbles J (1979) Ultrastructure of the interaction between Pyrenophora teres and a susceptible barley host. Can J Bot 57:40–47CrossRefGoogle Scholar
  47. Vincent D, Balesdent MH et al (2009) Hunting down fungal secretomes using liquid-phase IEF prior to high resolution 2-DE. Electrophoresis 30:4118–4136PubMedCrossRefGoogle Scholar
  48. Wallwork H (2000) ‘Cereal leaf and stem diseases’, 2 edn. Grains Research and Development Corporation, CanberraGoogle Scholar
  49. Wang X, Li X, Li Y (2007) A modified Coomassie brilliant blue staining method at nanogram sensitivity compatible with proteomic analysis. Biotechnol Lett 29:1599–1603PubMedCrossRefGoogle Scholar
  50. Zadoks J, Chang CTT, Konzak CF (1974) A decimal code for the growth stages of cereal. Weed Res 14:415–421CrossRefGoogle Scholar

Copyright information

© Australasian Plant Pathology Society Inc. 2014

Authors and Affiliations

  1. 1.School of Agriculture, Food & WineThe University of Adelaide, Waite Research InstituteGlen OsmondAustralia

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