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E ucalyptus P ine P athogen I nteractions

E ucalyptus P ine P athogen I nteractions. Introduction. Forest trees Long – lived Exposed to array of pathogens Do not posses adaptive immunity Innate immunity Resistant genotypes required. What do we know?. Arabidopsis thaliana. Pieterse et al 2009. P. P. Plant Defence.

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E ucalyptus P ine P athogen I nteractions

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  1. EucalyptusPine Pathogen Interactions

  2. Introduction • Forest trees • Long – lived • Exposed to array of pathogens • Do not posses adaptive immunity • Innate immunity • Resistant genotypes required

  3. What do we know? Arabidopsis thaliana Pieterse et al 2009

  4. P P Plant Defence CROSSLINKING OF CELL WALL PROTEINS O2 O2 K+ Cl- H2O2 Insect Pest Oomycete NADPH OXIDASE Volatiles Bacterial PATHOGEN KINASES PHOSPHATASES ROS H+ ELICITORS Fungal Ca2+ PAPILLAE MAPK CASCADES H2O2 R genes PHYTOALEXIN ACCUMULATION ROS JA ET SA Herbivory Cell wall fragments Secretions in oviposition fluid HR RESPONSE NPR1 2ndry metabolites e.g. alkaloids TRANSCRITION FACTORS ACTIVATED DEFENCE GENE ACTIVATION ACCUMULATION OF DEFENCE PROTEINS Proteinase inhibitors PPO LOX LOX, PAL, HEL, PDF 1.2 PR 1, 2, & 5

  5. Strategy • Information based on model systems • Arabidopsis thaliana • Develop pathosystems • Use genomic and transcriptomic resources • Dissect defence responses • Understand host responses • Identify key targets to improve resistance.

  6. P P CROSSLINKING OF CELL WALL PROTEINS O2 O2 K+ Cl- H2O2 Insect Pest Oomycete NADPH OXIDASE Volatiles Bacterial PATHOGEN KINASES PHOSPHATASES ROS H+ ELICITORS Fungal Ca2+ PAPILLAE MAPK CASCADES H2O2 R genes PHYTOALEXIN ACCUMULATION ROS JA ET SA Herbivory Cell wall fragments Secretions in oviposition fluid HR RESPONSE NPR1 2ndry metabolites e.g. alkaloids TRANSCRITION FACTORS ACTIVATED DEFENCE GENE ACTIVATION ACCUMULATION OF DEFENCE PROTEINS Proteinase inhibitors PPO LOX LOX, PAL, HEL, PDF 1.2 PR 1, 2, & 5

  7. A steptowardsunderstanding the SA and JA signalingpathways in E. grandis R. Naidoo, Dr. S. Naidoo, Prof AA. Myburg and Prof DK. Berger

  8. Eucalyptus Eucalyptus rust Paper Production Essential Oils Insect Invasion Eucalyptus Stem Cankers Timber http://scheererbearing.com/uploads/images/feature_pulppaper.jpg; http://topnews.in/health/files/Eucalyptus11.jpg; http://www.scielo.br/img/revistas/gmb/2010nahead/223fig01.jpg http://www.fs.fed.us/r5/spf/fhp/hawaii/eucalyptus%20rust%20on%20ohia.JPG; http://wiki.trin.org.au/pub/Wasps/LeptocybeInvasa/gallformer.jpg; http://www.wood-report.de/bilder/clonal_eucalyptus.jpg; http://2.bp.blogspot.com/_3uQiDL3c53s/Rx5nNpTvf0I/AAAAAAAAAWo/5ubDi6mK9ZQ/s400/Eucalyptus-In-Brazil-Celso-Foelkel-01.jpg;

  9. Plant Defence • Model Organism • Arabidopsis thaliana http://www.dreamstime.com/puzzle-with-missing-pieces-thumb5179822.jpg http://www-ijpb.versailles.inra.fr/en/sgap/equipes/cyto/images/Planche.gif

  10. P P Plant Defence CROSSLINKING OF CELL WALL PROTEINS O2 O2 K+ Cl- H2O2 Insect Pest Eucalyptus plant defence Oomycete NADPH OXIDASE Volatiles Bacterial PATHOGEN KINASES PHOSPHATASES ROS H+ ELICITORS Fungal Ca2+ PAPILLAE MAPK CASCADES H2O2 R genes PHYTOALEXIN ACCUMULATION ROS JA ET SA Herbivory Cell wall fragments Secretions in oviposition fluid HR RESPONSE NPR1 2ndry metabolites e.g. alkaloids TRANSCRITION FACTORS ACTIVATED DEFENCE GENE ACTIVATION ACCUMULATION OF DEFENCE PROTEINS Proteinase inhibitors PPO LOX LOX, PAL, HEL, PDF 1.2 PR 1, 2, & 5

  11. AIM Identify and characterize putative orthologs for defence marker genes (SA and JA) in E. grandis

  12. 1. Discovery of marker genes Gene Discovery of defence marker genes Phylogenetic trees Primer Design & PCR www.phytozome.net Verification of product

  13. Bioinformatics Pipeline

  14. EgrPR5 WAG + I + G I = 0.105 G = 1.195

  15. Verification of Genes

  16. 2. Dose Response

  17. SA - EgrPR2 * * * Students T-test P < 0.05

  18. MeJA - EgrPR4 *** *** Students T-test P < 0.001

  19. SA and JA antagonism SA JA JA SA Does this antagonistic relationship exist in Eucalyptus?

  20. Specificity of marker genes SA Marker + MeJA treated tissue * * * [MeJA] MeJA Marker + SA treated tissue * * [SA] * Students T-test P < 0.05

  21. 3. Time Course

  22. SA - EgrPR2 ** ** ** ** Students T-test P < 0.01

  23. MeJA - EgrPR4 * * ** *** Students T-test * P < 0.05, ** P < 0.01, *** P < 0.001

  24. 4. Infection trial

  25. TAG5 ZG14 Infection trial results TAG 5 lesion ZG14 lesion EgrPR2 ** *** * EgrPR4 * * * * Students T-test * P < 0.05; ** P < 0.01, *** P < 0.001

  26. How do the pieces fit together?

  27. Gene ortholog pipeline • Candidate defence genes • Identified specific concentrations • Elucidated window periods for improving defence • Diagnostic marker genes

  28. Conclusion • Eucalyptus “puzzle” • Possible diagnostic markers for plant defence • Elucidated the possible role of SA in defence against C. austroafricana • Stepping Stone in understandingEucalyptus – pathogen interactions http://www.pureskinjunkie.com/sitebuildercontent/sitebuilderpictures/greenLeafPuzzle.jpg

  29. Future Vision

  30. Acknowledgments • Supervisors and Fellow students • Prof Jolanda Roux • Adri Veale and GrietaMahlangu EPPI @

  31. Thank You

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