Trees

, Volume 26, Issue 1, pp 67–73 | Cite as

Real time qPCR expression analysis of some stress related genes in leaf tissue of Pyrus communis cv. Conférence after infection with Erwinia amylovora

Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

Hydrogen peroxide plays a critical role in the expression of disease resistance in several plant/pathogen interactions. It serves as a substrate for oxidative cross-linking of various plant cell wall components leading to the reinforcement of the cell structure, as a direct toxin against the pathogen and as a signal molecule for the induction of defence-related genes in the adjacent, still healthy tissues. In plant cells, enzymes and redox metabolites act in synergy to carry out the detoxification of hydrogen peroxide and other reactive oxygen species (ROS). Superoxide dismutase (SOD) catalyses the dismutation of superoxide to hydrogen peroxide, catalase (CAT) dismutates hydrogen peroxide to oxygen and water, and ascorbate peroxidase (APX) reduces hydrogen peroxide to water by utilising ascorbate as specific electron donor. These are considered some of the main enzymatic systems for protecting cells against oxidative damage. These enzymes are present in various isozyme forms in several cell compartments and their expression is genetically controlled and regulated both by developmental and environmental stimuli, according to the necessity to remove ROS produced in cells. The aim of this study was to determine the possible role of these antioxidants in the defence mechanism of Pyrus communis cv. Conférence leaf tissue after an infection with Erwinia amylovora. Shoots of 2-year-old pear trees cv. Conférence were infected with E. amylovora strain SGB 225/12, were mock infected or left untreated. To account for structural changes, not only a difference was made between control, infected and mock-infected leaves, but we also included a distinction between young and old leaves, because it is known that older leaves are less susceptible for fire blight infections. Leaf samples were taken at specific time points after infection and the expression pattern of not necrotic tissue close to the infection site was analysed for their diverse isoforms of SOD, APX and CAT by using real time qPCR. In this study, no striking differences in transcription patterns of these enzymes between control, mock infected and E. amylovora infected leaves were observed. However, a significant difference between the expression levels of some genes in young and old leaves was noticed. These differences could partially explain the different progression rate by which E. amylovora infects, invades and causes necrosis in young and old leaves.

Keywords

Erwinia amylovora Pear Oxidative stress Plant defence Real time qPCR Leaf ontogenesis 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

This work was conducted within the framework of the European Research Network COST864.

References

  1. Alscher RG, Donahue JL, Cramer CL (1997) Reactive oxygen species and antioxidants: relationships in green cells. Physiol Plant 100:224–233CrossRefGoogle Scholar
  2. Andreotti C, Costa G, Treutter D (2006) Composition of phenolic compounds in pear leaves as affected by genetics, ontogenesis and the environment. Sci Hortic 109:130–137CrossRefGoogle Scholar
  3. Arora A, Sairam RK, Srivastava GC (2002) Oxidative stress and antioxidative system in plants. Curr Sci 82:1227–1238Google Scholar
  4. Baker CJ, Orlandi EW (1995) Active oxygen in plant pathogenesis. Annu Rev Phytopathol 33:299–321CrossRefPubMedGoogle Scholar
  5. Bent AF, Innes RW, Ecker JR, Staskawicz BJ (1992) Disease development in ethylene-insensitive Arabidopsis-thaliana infected with virulent and avirulent Pseudomonas and Xanthomonas pathogens. Mol Plant Microbe Interact 5:372–378CrossRefPubMedGoogle Scholar
  6. Blokhina O, Virolainen E, Fagerstedt KV (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot 91:179–194CrossRefPubMedGoogle Scholar
  7. Bolwell GP, Wojtaszek P (1997) Mechanisms for the generation of reactive oxygen species in plant defence—a broad perspective. Physiol Mol Plant Pathol 51:347–366CrossRefGoogle Scholar
  8. Bringe K, Schumacher CFA, Schmitz-Eiberger M, Steiner U, Oerke EC (2006) Ontogenetic variation in chemical and physical characteristics of adaxial apple leaf surfaces. Phytochemistry 67:161–170CrossRefPubMedGoogle Scholar
  9. Buchanan BB, Balmer Y (2005) Redox regulation: a broadening horizon. Ann Rev Plant Biol 56:187–220CrossRefGoogle Scholar
  10. Chamnongpol S, Willekens H, Moeder W, Langebartels C, Sandermann H, Van Montagu A et al (1998) Defense activation and enhanced pathogen tolerance induced by H2O2 in transgenic tobacco. Proc Natl Acad Sci USA 95:5818–5823CrossRefPubMedGoogle Scholar
  11. Dat J, Vandenabeele S, Vranova E, Van Montagu M, Inze D, Van Breusegem F (2000) Dual action of the active oxygen species during plant stress responses. Cell Mol Life Sci 57:779–795CrossRefPubMedGoogle Scholar
  12. Deckers T, Schoofs H (2008) Status of the pear production in Europe. Acta Hortic 800:95–105Google Scholar
  13. Develey-Riviere MP, Galiana E (2007) Resistance to pathogens and host developmental stage: a multifaceted relationship within the plant kingdom. New Phytol 175:405–416CrossRefPubMedGoogle Scholar
  14. Fischer TC, Gosch C, Pfeiffer J, Halbwirth H, Halle C, Stich K et al (2007) Flavonoid genes of pear (Pyrus communis). Trees—Struct Funct 21:521–529CrossRefGoogle Scholar
  15. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875CrossRefPubMedGoogle Scholar
  16. Gasic K, Hernandez A, Korban SS (2004) RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction. Plant Mol Biol Report 22:437a–437gCrossRefGoogle Scholar
  17. Gayler S, Leser C, Priesack E, Treutter D (2004) Modelling the effect of environmental factors on the “trade-off” between growth and defensive compounds in young apple trees. Trees—Struct Funct 18:363–371CrossRefGoogle Scholar
  18. Heyens K, Valcke R, Dumont D, Robben J, Noben JP (2006) Differential expression of proteins in apple following inoculation with Erwinia amylovora. Acta Hortic 704:69–74Google Scholar
  19. Huttner C, Beuerle T, Scharnhop H, Ernst L, Beerhues L (2010) Differential effect of elicitors on biphenyl and dibenzofuran formation in Sorbus aucuparia cell cultures. J Agric Food Chem 58:11977–11984CrossRefPubMedGoogle Scholar
  20. Kus JV, Zaton K, Sarkar R, Cameron RK (2002) Age-related resistance in Arabidopsis is a developmentally regulated defense response to Pseudomonas syringae. Plant Cell 14:479–490CrossRefPubMedGoogle Scholar
  21. Li B, Xu X (2002) Infection and development of apple scab (Venturia inaequalis) on old leaves. J Phytopathol-Phytopatholog Zeitschrift 150:687–691CrossRefGoogle Scholar
  22. Lim PO, Kim HJ, Nam HG (2007) Leaf senescence. Ann Rev Plant Biol 58:115–136CrossRefGoogle Scholar
  23. Mayr U, Treutter D, Santosbuelga C, Bauer H, Feucht W (1995) Developmental-changes in the phenol concentrations of golden delicious apple fruits and leaves. Phytochemistry 38:1151–1155CrossRefPubMedGoogle Scholar
  24. Milcevicova R, Gosch C, Halbwirth H, Stich K, Hanke MV, Peil A et al (2010) Erwinia amylovora-induced defense mechanisms of two apple species that differ in susceptibility to fire blight. Plant Sci 179:60–67CrossRefGoogle Scholar
  25. Navrot N, Rouhier N, Gelhaye E, Jacquot JP (2007) Reactive oxygen species generation and antioxidant systems in plant mitochondria. Physiol Plant 129:185–195CrossRefGoogle Scholar
  26. Nicaise V, Roux M, Zipfel C (2009) Recent advances in PAMP-triggered immunity against bacteria: pattern recognition receptors watch over and raise the alarm. Plant Physiol 150:1638–1647CrossRefPubMedGoogle Scholar
  27. Robert-Seilaniantz A, Navarro L, Bari R, Jones JD (2007) Pathological hormone imbalances. Curr Opin Plant Biol 10:372–379CrossRefPubMedGoogle Scholar
  28. ThordalChristensen H, Zhang ZG, Wei YD, Collinge DB (1997) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley–powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  29. Venisse JS, Gullner G, Brisset MN (2001) Evidence for the involvement of an oxidative stress in the initiation of infection of pear by Erwinia amylovora. Plant Physiol 125:2164–2172CrossRefPubMedGoogle Scholar
  30. Venisse JS, Barny MA, Paulin JP, Brisset MN (2003) Involvement of three pathogenicity factors of Erwinia amylovora in the oxidative stress associated with compatible interaction in pear. FEBS Lett 537:198–202CrossRefPubMedGoogle Scholar
  31. Whalen MC (2005) Host defence in a developmental context. Mol Plant Pathol 6:347–360CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • K. Vrancken
    • 1
  • H. Schoofs
    • 2
  • T. Deckers
    • 2
  • R. Valcke
    • 1
  1. 1.Molecular and Physical Plant PhysiologyHasselt UniversityDiepenbeekBelgium
  2. 2.Pomology DepartmentPC Fruit Research StationSint-TruidenBelgium

Personalised recommendations