1. Unravelling host plant resistance in chrysanthemum using NMR Suzanne Kos, MSc

2. Introduction Plant pests Integrated Pest Management Host plant resistance

3. Ecometabolomic approach –Bioassays to determine resistance –Metabolomics to identify metabolites –In vitro bioassays for confirmation Western flower thrips –Polyphagous pest –Silver and growth damage –Transmission of plant viruses Introduction

4. Ecometabolomic approach Senecio (Leiss et al. 2009) –PAs jacobine, jaconine N-oxide –The flavanoid kaempferol glucoside Tomato (Mirnezad et al. 2009) –Acylsugars Carrot (Leiss et al. 2013) –The flavanoid luteolin –The phenylpropanoid sinapic acid –The amino acid beta-alanine

5. Chrysanthemum Important Dutch greenhouse ornamental Pest problem: western flower thrips, celery leafminer and two-spotted spider mite Leiss et al. (2009) identified chlorogenic acid as resistance factor for thrips

6. 1. In-vivo bioassay with 73 cultivars in greenhouse 2. In-vivo bioassays with 12 cultivars in climate room 3. Identification and quantification of metabolites with NMR 4. Cross reference of resistance by in-vitro bioassay Ecometabolomic approach

7. 1. In-vivo bioassay with 73 cultivars in greenhouse 73 cultivars, 5 replicates each Five adult thrips per plant released Scored thrips silver damage after 3 ½ weeks

8. 1. In-vivo bioassay with 73 cultivars in greenhouse 5 replicates per cultivar 5 adult thrips per plant released Scored thrips silver damage after 3 ½ weeks F=7.571, df=72, p<0.001

9. 1. In-vivo bioassay with 73 cultivars in greenhouse 5 replicates per cultivar 5 adult thrips per plant released Scored thrips silver damage after 3 ½ weeks F=7.571, df=72, p<0.001

10. 2. In-vivo bioassays with 12 cultivars in climate room ANOVA: F=9.848, df=11, p<0.001

11. 2. In-vivo bioassays with 12 cultivars in climate room Twenty unsexed adult thrips per leaf in Petri dish for 24h. All larvae that emerged from eggs were counted.

12. ANOVA: F=3.496, df=8, P=0.005 2. In-vivo bioassays with 12 cultivars in climate room

13. Pearson correlation: R=0.772, p=0.007

14. Different protons in a molecule resonate at slightly different frequencies when placed in a magnetic field depending upon the details of the electron motion in the nearby atoms. Structural information about molecules Easy sample preparation and highly reproducable Quantification Nuclear Magnetic Resonance 14 PhenolicsCarbohydrates, PAs Amino acids, terpenoids

15. 3. Identification and quantification of metabolites with NMR

16. Resistant Susceptible R2=91,4% Q2=68,5%

17. 3. Identification and quantification of metabolites with NMR Resistant Susceptible

18. 3. Identification and quantification of metabolites with NMR

19. ANOVA: F=4.028, df=11, P=0.001

20. 3. Identification and quantification of metabolites with NMR Spearman: ρ=-0.538, N=12, P=0.035

21. 1. In-vivo bioassay with 73 cultivars in greenhouse 2. In-vivo bioassays with 12 cultivars in climate room 3. Identification and quantification of metabolites with NMR 4. Cross reference of resistance by in-vitro bioassay Ecometabolomic approach

22. 4. Cross reference of resistance by in-vitro bioassay

23. Acknowledgements C. Hermans Dr. K.A. LeissProf. P. Klinkhamer Dr. Y.H. Choi Plant Ecology and Phytochemistry group: Funding provided by: In cooperation with: