Functional & Integrative Genomics

, Volume 10, Issue 4, pp 619–627 | Cite as

Lipid transfer proteins and protease inhibitors as key factors in the priming of barley responses to Fusarium head blight disease by a biocontrol strain of Pseudomonas fluorescens

Short Communication


Strains of non-pathogenic pseudomonad bacteria, can elicit host defence responses against pathogenic microorganisms. Pseudomonas fluorescens strain MKB158 can protect cereals from pathogenesis by Fusarium fungi, including Fusarium head blight which is an economically important disease due to its association with both yield loss and mycotoxin contamination of grain. Using the 22 K barley Affymetrix chip, trancriptome studies were undertaken to determine the local effect of P. fluorescens strain MKB158 on the transcriptome of barley head tissue, and to discriminate transcripts primed by the bacterium to respond to challenge by Fusarium culmorum, a causal agent of the economically important Fusarium head blight disease of cereals. The bacterium significantly affected the accumulation of 1203 transcripts and primed 74 to positively, and 14 to negatively, respond to the pathogen (P = 0.05). This is the first study to give insights into bacterium priming in the Triticeae tribe of grasses and associated transcripts were classified into 13 functional classes, associated with diverse functions, including detoxification, cell wall biosynthesis and the amplification of host defence responses. In silico analysis of Arabidopsis homologs of bacterium-primed barley genes indicated that, as is the case in dicots, jasmonic acid plays a role in pseudomonad priming of host responses. Additionally, the transcriptome studies described herein also reveal new insights into bacterium-mediated priming of host defences against necrotrophs, including the positive effects on grain filling, lignin deposition, oxidative stress responses, and the inhibition of protease inhibitors and proteins that play a key role in programmed cell death.


Aquaporins Cell wall Chymotrypsin Gibberellins Induced resistance Jasmonic acid nsLTP Serpins Seed storage proteins 



The authors wish to thank Mr. Gerard Leonard for his help with plant propagation, Mr. Damian Egan for technical assistance and Dr. Kathrin Reiber for reading the manuscript. This research was funded by the Irish Department of Agriculture and Fisheries Research Stimulus Fund 2006 (RSF 06 377).

Supplementary material

10142_2010_177_MOESM1_ESM.doc (20 kb)
ESM 1 (DOC 32 kb)
10142_2010_177_MOESM2_ESM.doc (50 kb)
Table S1 Sequence of transcript-specific primers used for real time RT-PCR analyses. (DOC 49 kb)
10142_2010_177_MOESM3_ESM.doc (19 kb)
Table S2 The effect of Pseudomonas fluorescens (strain MKB158), as compared to Fusarium culmorum (strain FCF200), on transcript accumulation in barley cultivar Lux head tissue at either 24 or 48 h post-pathogen treatment, as determined by microarray analysis. (DOC 18 kb)
10142_2010_177_MOESM4_ESM.doc (222 kb)
Table S3 Description of the complementary potentiated transcripts not included in Table 1. Functional classification of barley (cultivar Lux) transcripts that were differentially regulated in head tissue in response to treatment with the biocontrol bacterium Pseudomonas fluorescens strain MKB158 and/or the pathogenic fungus Fusarium culmorum strain FCF200 at 24 or 48 h post-pathogen treatment a . (DOC 221 kb)
10142_2010_177_MOESM5_ESM.doc (55 kb)
Fig. S1 Venn diagrams highlighting the influence of the treatment with the bacterium Pseudomonas fluorescens (strain MKB158) on transcript accumulation in flowering heads of barley cultivar Lux in the absence and presence of the pathogen Fusarium culmorum (strain FCF200), as determined by microarray analysis. Diagrams represent the numbers of contigs (A) up-regulated and (B) down-regulated at 24 and/or 48 h post-pathogen treatment (pathogen was applied 24 h post-bacterium treatment). Numbers indicate the amount of barley contigs with significantly altered expression (P = 0.05) that are either shared or exclusive to bacterium or pathogen treatment. Codes: B, bacterium (P. fluorescens); P, pathogen (F. culmorum). (DOC 55 kb)
10142_2010_177_MOESM6_ESM.xls (818 kb)
Supplemental Expression Data File Expression values (signals) detected for select probes of interesta on the barley affymetrix chip in barley head tissue in response to treatment with bacterium Pseudomonas fluorescens strain MKB158 (B), the fungal pathogen Fusarium culmorum strain FCF200 (P) or both agents in three experiments (Expt 1, Expt 2 and Expt 3) at either 24 or 48 h post-pathogen treatment. For each probe, detection signal and associated statistical significance (P values) are given. (XLS 817 kb)


  1. Almagro L, Gomez Ros LV, Belchi-Navarro S, Bru R, Ros Barcelo A, Pedreno MA (2009) Class III peroxidase in plant defence reactions. J Exp Bot 60(2):377–390CrossRefPubMedGoogle Scholar
  2. Bakker PA, Pieterse CM, Van Loon L (2007) Induced systemic resistance by fluorescent Pseudomonas spp. Phytopathology 97:239–243CrossRefPubMedGoogle Scholar
  3. Bertani G (1951) Studies on lysogenesis. I. The mode of phage liberation by lysogenic Echerichia coli. J Bacteriol 62(3):292–300Google Scholar
  4. Cartieaux F, Thibaud MC, Zimmerli L, Lessard P, Sarrobert C, David P, Gerbaud A, Robaglia C, Somerville S, Nussaume L (2003) Transcriptome analysis of Arabidopsis colonized by a plant-growth promoting rhizobacterium reveals a general effect on disease resistance. Plant J 36(2):177–188CrossRefPubMedGoogle Scholar
  5. Cartieaux F, Contesto C, Gallou A, Desbrosses G, Kopta J, Taconnat L, Renou JP, Touraine B (2008) Simultaneous interaction of Arabidopsis thaliana with Bradyrhizobium sp. ORS278 and Pseudomonas syringae pv. tomato DC3000 leads to complex transcriptome changes. Mol Plant-Microbe Interact 21(2):244–259CrossRefPubMedGoogle Scholar
  6. Chang S, Puryer J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116CrossRefGoogle Scholar
  7. Compant S, Duffy B, Nowak J, Clement C, Ait Barka E (2005) Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action and future prospects. Appl Environ Microbiol 71(9):4951–4959CrossRefPubMedGoogle Scholar
  8. Compier M (2005) Hemicellulose biosynthesis and degradation in tobacco cell walls. Dissertation, Wageningen University, The NetherlandsGoogle Scholar
  9. Daniels MJ, Yeager M (2005) Phosphorylation of aquaporin PvTIP3;1 defined by mass spectrometry and molecular modeling. Biochemistry 44:14443–14454CrossRefPubMedGoogle Scholar
  10. Davin LB, Wang HB, Crowell AL, Bedgar DL, Martin DM, Sarkanen, Lewis NG (1997) Stereoselective bimolecular phenoxy radical coupling by an auxillary (dirigent) protein without an active center. Science 275:362–366CrossRefPubMedGoogle Scholar
  11. Desmond OJ, Manners JM, Stephens AE, Maclean DJ, Schenk PM, Gardiner DM, Munn AL, Kazan K (2008) The Fusarium mycotoxin deoxynivalenol elicts hydrogen peroxide production, programmed cell death and defence responses in wheat. Mol Plant Pathol 9(4):435–445CrossRefPubMedGoogle Scholar
  12. Fleming JM, Long EL, Ginsburg E, Gerscovich D, Metzer PS, Vonderhaar BK (2008) Interlobular and intralobular mammay stroma: genotype may not reflect phenotype. BMC Cell Biol. doi: 10.1186/1471-2121-9-46 PubMedGoogle Scholar
  13. Jiang C, Fu X (2007) GA action: turning on de-DELLA repressing signalling. Curr Opin Plant Biol 10(5):461–465CrossRefPubMedGoogle Scholar
  14. Jiang F, Zheng X, Chen J (2009a) Microarray analysis of gene expression profile induced by the biocontrol yeast Cryptococcus laurentii in cherry tomato fruit. Gene 430(1–2):12–16CrossRefGoogle Scholar
  15. Jiang F, Chen JS, Miao Y, Krupinska K, Zheng XD (2009b) Identification of differentially expressed genes from cherry tomato fruit (Lycopersicon esculentum) after application of the biological control yeast Cryptococcus laurentii. Postharvest Biol Technol 53(3):131–137CrossRefGoogle Scholar
  16. Johansson A, Rasmussen SK, Harthill JE, Welinder KG (1992) cDNA, amino acid and carbohydrate sequence of a barley seed-specific peroxidase BP1. Plant Mol Biol 18(6):1151–1161CrossRefPubMedGoogle Scholar
  17. Johnson KD, Hofte H, Chrispeels MJ (1990) An intrinsic tonoplast protein of protein storage vacuoles in seeds is structurally related to a bacterial solute transporter (GlpF). Plant Cell 2:525–532CrossRefPubMedGoogle Scholar
  18. Keates SE, Kostman TA, Anderson JD, Bailey BA (2003) Altered gene expression in three plant species in response to treatment with Nep1, a fungal protein that causes necrosis. Plant Physiol 132(3):1610–1622CrossRefPubMedGoogle Scholar
  19. Khan MR, Doohan FM (2009) Bacterium-mediated control of Fusarium head blight disease of wheat and barley and associated mycotoxin contamination of grain. Biol Control 48(1):42–47CrossRefGoogle Scholar
  20. Khan MR, Fischer S, Egan D, Doohan FM (2006) Biological control of fusarium seedling blight disease of wheat and barley. Phytopathology 96(4):386–394CrossRefPubMedGoogle Scholar
  21. La Camera S, Geoffroy P, Samaha H, Ndiaye A, Rahim G, Legrand M, Heitz T (2005) A pathogen-inducible patatin-like lipid acyl hydrolase facilitates fungal and bacterial host colonization in Arabidopsis. Plant J 44:810–825CrossRefPubMedGoogle Scholar
  22. La Camera S, Balague C, Gobel C, Geoffroy P, Legrand M, Feussner I, Roby D, Heitz T (2009) The Arabidopsis patatin-like protein 2 (PLP2) plays an essential role in cell death execution and differentially affects biosynthesis of oxylipins and resistance to pathogens. Mol Plant-Microbe Interact 22:469–481CrossRefPubMedGoogle Scholar
  23. Liu G, Sheng X, Greenshields DL, Ogieglo A, Kaminskyj S, Selvaraj G, Wei Y (2005) Profiling of wheat class III peroxidase genes derived from mildew-attacked epidermis reveals distinct sequence-associated expression patterns. Mol Plant Microbe Interact 18(7):730–741CrossRefPubMedGoogle Scholar
  24. Livak KJ, Schmittegen TD (2001) Analysis of relative gene expression data using real-time quantitave PCR and the 2-ΔΔCt method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  25. Maldonado AM, Doerner P, Dixon RA, Lamb CJ, Cameron RK (2002) A putative lipid transfer protein involved in systemic resistance signalling in Arabidopsis. Nature 419(6905):399–403CrossRefPubMedGoogle Scholar
  26. Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9(10):490–498CrossRefPubMedGoogle Scholar
  27. Morey JS, Ryan JC, Van Dolah FM (2006) Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR. Biol Proced Online 8(1):175–193CrossRefPubMedGoogle Scholar
  28. Navarro L, Bari R, Achard P, Lison P, Nemri A, Harberd NP, Jones JDG JDG (2008) DELLAs control plant immune responses by modulating the balance of jasmonic acid and salicylic acid signaling. Curr Biol 18(9):650–655CrossRefPubMedGoogle Scholar
  29. Nehme A, Lobenhofer EK, Stamer WD, Edelman JL (2009) Glucocorticoids with different chemical structures but similar glucocorticoid receptor potency regulate subsets of common and unique genes in human trabecular meshwork cells. BMC Med Genomics. doi: 10.1186/1755-8794/2/58 PubMedGoogle Scholar
  30. Onodera Y, Suzuki A, Wu CY, Washida H, Takaiwa F (2001) A rice functional transcriptional activator, RISBZ1, responsible for endosperm-specific expression of storage protein genes through GCN4 motif. J Biol Chem 276(17):14139–14152PubMedGoogle Scholar
  31. Parry DW, Jenkinson P, McLeod L (1995) Fusarium ear blight (scab) in small grain cereals—a review. Plant Pathol 44:207–238CrossRefGoogle Scholar
  32. Pekkarinen AI, Jones BL (2002) Trypsin-like proteinase produced by Fusarium culmorum grown on grain proteins. J Agric Food Chem 50:3849–3855CrossRefPubMedGoogle Scholar
  33. Pekkarinen AI, Longstaff C, Jones BL (2007) Kinetics of the inhibition of Fusarium serine proteinases by barley (Hordeum vulgare L.) inhibitors. J Agric Food Chem 55(7):2736–2742CrossRefPubMedGoogle Scholar
  34. Petersen ML, Hejgaard J, Thompson GA, Schulz A (2005) Cucurbit phloem serpins are graft-transmissible and appear to be resistant to turnover in the sieve element-companion cell complex. J Exp Bot 56(422):3111–3120CrossRefGoogle Scholar
  35. Pieterse CMJ, van Wees SCM, van Pelt JA, Knoester M, Laan R, Gerrits H, Weisbeek PJ, van Loon LC (1998) A novel signaling pathway controlling induced systemic resistance in arabidopsis. Plant Cell 10:1571–1580CrossRefPubMedGoogle Scholar
  36. Pieterse CM, Leon-Reyes A, Van der Ent S, Van Wees SCM (2009) Networking by small-molecule hormones in plant immunity. Nat Chem Biol 5(5):308–316CrossRefPubMedGoogle Scholar
  37. Roberts TH, Hejgaard J (2008) Serpins in plants and green algae. Funct Integ Genomics 8(1):1–27CrossRefGoogle Scholar
  38. Salcedo G, Sanchez-Monge R, Barber D, Diaz-Perales A (2007) Plant non-specific lipid transfer proteins: an interface between plant defence and human allergy. Biochim Biophys Acta-Mol Cell Biol Lipids 1771(6):781–791Google Scholar
  39. Sarosh BR, Danielsson J, Meijer J (2009) Transcript profiling of oilseed rape (Brassica napus) primed for biocontrol differentiate genes involved in microbial interactions with beneficial Bacillus amyloliquefaciens from pathogenic Botrytis cinerea. Plant Mol Biol 70(1–2):31–45CrossRefPubMedGoogle Scholar
  40. Schwechheimer C (2008) Understanding gibberellic acid signaling—are we there yet? Curr Opin Plant Biol 11(1):9–15CrossRefPubMedGoogle Scholar
  41. Simonetti EP, Veronico M, Melillo T, Delibes A, Fe Andres M, Lopez-Brana I (2009) Analysis of class III peroxidase genes expressed in roots of resistant and susceptible wheat lines infected by Heterodera avenae. Mol Plant Microbe Interact 22(9):1081–1092CrossRefPubMedGoogle Scholar
  42. Smirnoff N, Grant M (2008) Do DELLAs do defence? Curr Biol 18(14):617–619CrossRefGoogle Scholar
  43. Sreenivasulu N, Graner A, Wobus U (2008) Barley Genomics: an overview. Int J Plant Genomics 2008:1–13CrossRefGoogle Scholar
  44. Van der Ent S, Van Hulten M, Pozo MJ, Czechowski T, Udvardi MK, Pieterse CMJ, Ton J (2009) Priming of plant innate immunity by rhizobacteria and β-aminobutyric acid: differences and similarities in regulation. New Phytol 183:419–431CrossRefPubMedGoogle Scholar
  45. Van Hulten M, Pelser M, Van Loon L, Pieterse CM, Ton J (2006) Costs and benefits of priming for defence in Arabidopsis. Proc Natl Acad Sci USA 103(14):5602–5607CrossRefPubMedGoogle Scholar
  46. Van Wees SCM, Luijendijk M, Smoorenburg I, Van Loon L, Pieterse CMJ (1999) Rhizobacteria-mediated induced systemic resistance (ISR) in Arabidopsis is not associated with a direct effect on gene expression of known defence-related genes but stimulates the expression of jasmonate-inducible gene atvsp upon challenge. Plant Mol Biol 41:537–549CrossRefPubMedGoogle Scholar
  47. Van Wees SCM, Van der Ent S, Pieterse CMJ (2008) Plant immune responses triggered by beneficial microbes. Curr Opin Plant Biol 11(4):443–448CrossRefPubMedGoogle Scholar
  48. Verhagen BWM, Glazebrook J, Zhu T, Chang HS, Van Loon L, Pieterse CMJ (2004) The transcriptome of rhizobacteria-induced systemic resistance in Arabidopsis. Mol Plant-Microbe Interact 17(8):895–908CrossRefPubMedGoogle Scholar
  49. Walter S, Brennan JM, Arunachalan C, Ansari KI, Hu X, Trognitz F, Trognitz B, Leonard G, Egan D, Doohan FM (2008) Components of the gene network associated with genotype-dependent response of wheat to the Fusarium mycotoxin deoxynivalenol. Funct Integ Genomics 8:421–427CrossRefGoogle Scholar
  50. Wang YQ, Ohara Y, Nakayashiki H, Tosa Y, Mayama S (2005) Microarray analysis of the gene expression profile induced by the endophytic plant growth-promoting rhizobacteria, Pseudomonas fluorescens FPT9601-T5 in Arabidopsis. Mol Plant-Microbe Interact 18(5):385–396CrossRefPubMedGoogle Scholar
  51. Willigen CV, Postaire O, Tournaire-Roux C, Bursiac Y, Maurel C (2006) Expression and inhibition of aquaporins in germinating Arabidopsis seeds. Plant Cell Physiol 47(9):1241–1250CrossRefGoogle Scholar
  52. Xue AG, Voldeng HD, Savard ME, Fedak G, Tian X, Hsiang T (2009) Biological control of fusarium head blight of wheat with Clonostachys rosea strain ACM941. Can J Plant Pathol 31(2):169–179CrossRefGoogle Scholar
  53. Zimmermann R, Werr R (2005) Pattern formation in the monocot embryo as revealed by NAM and CUC3 orthologues from Zea mais L. Plant Mol Biol 58:669–685CrossRefPubMedGoogle Scholar
  54. Zimmermann P, Hirsh-Hoffmann M, Henning L, Gruissem W (2004) GENEVESTIGATOR. Arabidopsis microarray database and analysis toolbox. Plant Physiol 136(1):2621–2632CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Carloalberto Petti
    • 1
  • Mojibur Khan
    • 1
  • Fiona Doohan
    • 1
  1. 1.School of Biology and Environmental Science, Molecular Plant-Pathogen Interaction Group, Science WestUniversity College DublinDublinIreland

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