1. Febé Meyer Dr. Sanushka Naidoo Prof. Zander Myburg Dr. Noelani van den Berg

2. Eucalyptus + Phytophthora Phytophthora destructive genus of hemi-biotrophs Cold-tolerant eucalypts: susceptible Root and collar rot, die-back Jolanda Roux

4. P. cinnamomi, 1922 Google earth ©2009 t2.gstatic.com

5. Phytophthora

6. E. nitens + P. cinnamomi InfectedControl 1 cm Compatible system: 1.Different stages of interaction 2.Snapshot of successful infection 3.Attempted defence by host Compatible system: 1.Different stages of interaction 2.Snapshot of successful infection 3.Attempted defence by host

8. Aim: To profile attempted defence mechanisms in E. nitens against P. cinnamomi Aim: To profile attempted defence mechanisms in E. nitens against P. cinnamomi

9. RNA quality Control 1 2 3 Inoculated 1 2 3 28S 18S

10. Sample name Total reads Mapped (%) Expressed genes Average FPKM Control 13744480976.6829024493598 Control 23611167878.1829250492972 Control 33706025177.6629135467006 Inoculated 13723437177.7829429471923 Inoculated 23662243479.7829407497171 Inoculated 33602297878.3729576473466 Mapping results

11. Differentially expressed

12. regulation of DNA- dependant transcripti on in response to stress detoxificati on of nitrogen compound defence response to bacterium, incompatib le interaction response to toxic substanc e defence response by callose depositio n phosphorel ay signal transductio n system plant-type cell wall loosening plant- type cell wall modif icatio n Syncytiu m formatio n heter ocycle metab olism monosacc haride biosynthes is alcohol biosynthesi s sulfur amino acid metabolism Overrepresentation: up-regulated Reactive oxygen species JA and SA signalling Flavonoid biosynthesis Water deprivation

13. Overrepresentation: down-regulated Photosynthesis Secondary cell wall Auxin & giberellin Lignin biosynthesis

14. PR-5Eucgr.D01888 Eucgr.E01381 Eucgr.E01382 Eucgr.E01384 Eucgr.E01389 Eucgr.E01385 Eucgr.D01887 Eucgr.D01892 Eucgr.E00557 Eucgr.E00560 Eucgr.H03863 Eucgr.H03864 Eucgr.H03865 Eucgr.L01962 Eucgr.B00944 Eucgr.G01772 Eucgr.H04405 Eucgr.J02061 PR-1 PR-3 PR-5 Up Down PR-1BEucgr.D01560 Eucgr.D01552 Eucgr.I02509 Eucgr.G01140 Eucgr.G01134 Eucgr.G01148 PR-1AEucgr.G01137 Eucgr.L01707 Eucgr.G01171 Eucgr.L02505 PR-1 likeEucgr.C02418 Eucgr.D01550 Eucgr.D01554 PR-3 Eucgr.I01495 Eucgr.I02271 Eucgr.J02519 Eucgr.L00938 Eucgr.L00937 Eucgr.J02518 Eucgr.L00941 Eucgr.L00939 PR-9Eucgr.A01385 Eucgr.F04198 Eucgr.I00717 Eucgr.D01857 Eucgr.F04195 Eucgr.J02352 Eucgr.L02740 Eucgr.E04056 Eucgr.F03724 Eucgr.J02173 PR-9 Papaya Soybean Potato Eucalyptus

15. Conclusion Growth down, defence up Attempted defence: ▫JA/ET signalling ▫PR genes ▫Flavonoids Possible manipulation/cause of susceptibility ▫Peroxidase putative ortholog ▫Lignin (avoid strengthening cell walls) ▫Flavonoids? – chalcone synthase down ▫Water stress

16. Future work Mapping back to P. cinnamomi to determine what pathogen genes are expressed Compare to responses in resistant/highly tolerant Eucalyptus spp.

17. Acknowledgements Supervisors Research groups: EPPI; FMG; MPPI; FTBP Sitha Ramsuchit

18. PR genes Eucalyptus ID log2 (fold change) TAIR IDTAIR description A. thaliana (Col-0) A. thaliana (Ler) PR-1 Eucgr.D015524.559 AT2G14580.1Basic pathogenesis-related protein 1 Eucgr.D015604.355 Eucgr.G011343.338 Eucgr.G011404.396 Eucgr.G011484.365 Eucgr.G011373.243 AT2G14610.1Pathogenesis-related gene 1++ Eucgr.G011714.307 Eucgr.L017073.131 Eucgr.L025053.142

19. PR genes Eucalyptus ID log2 (fold change) TAIR IDTAIR description Persea america na Zea mays PR-3 - Chitinase class I, II, IV, VI, VII Eucgr.L009414.016 AT3G12500.1Basic chitinase++ Eucgr.J025193.936 Eucgr.L009383.930 Eucgr.L009393.734 Eucgr.L009373.465 Eucgr.I014953.110 Eucgr.H04034-1.329 AT3G16920.1Chitinase-like protein 2+ Eucgr.H00455-0.754 AT1G05850.1Chitinase family protein (TAIR 9) Eucgr.K021662.527 AT3G54420.1Homolog of carrot EP3-3 chitinase++ Eucgr.K021662.208 Eucgr.H003263.754 Eucgr.H003213.751 Eucgr.H003283.569 Eucgr.A000201.434

20. PR genes Eucalyptus ID log2 (fold change) TAIR IDTAIR description Persea america na Zea mays PR-5 - Thaumatin-like and osmotin Eucgr.H038635.086 AT4G11650.1Osmotin 34 + Eucgr.H038654.931 Eucgr.H038644.775 Eucgr.L019624.693 Eucgr.E005574.433 Eucgr.D018883.574 Eucgr.D018923.342 Eucgr.D018873.306 Eucgr.E005602.968 Eucgr.A014741.031 AT5G38280.1PR5-like receptor kinase Eucgr.A014700.964 Eucgr.A014780.776 Eucgr.E013825.424 AT1G20030.2Pathogenesis-related thaumatin superfamily protein + Eucgr.E013845.310 Eucgr.E013895.225 Eucgr.E013855.129 Eucgr.E013814.955 Eucgr.B00944-1.358 AT1G73620.1 Pathogenesis-related thaumatin superfamily protein Eucgr.J02061-1.182 AT2G28790.1 ++ Eucgr.G01772-1.330 AT4G38660.1

21. PR genes Eucalyptus ID log2 (fold change) TAIR IDTAIR description Persea america na PR-9 - Peroxidase Eucgr.D018573.077 AT4G11600.1Glutathione peroxidase 6 (TAIR 9) + Eucgr.F041982.343 AT1G71695.1Peroxidase superfamily protein + Eucgr.F041951.462 Eucgr.L027401.389 Eucgr.J023523.654 AT4G37530.1 Peroxidase superfamily protein + Eucgr.A013851.834 AT1G05260.1 + Eucgr.J02173-1.229 AT5G40150.1 Eucgr.F03724-1.760 AT5G42180.1 Eucgr.E04056-1.850 AT4G21960.1 +