1. Rabea Schweiger 1,2, Anna-Maria Heise 1, Marcus Persicke 2, Caroline Müller 1,2 1 Department of Chemical Ecology, 1,2 Center for Biotechnology (CeBiTec), Bielefeld University Specificity of metabolic plant responses to salicylic and jasmonic acid and impacts on chewing and sucking herbivores

2. Herbivores Piercing- sucking Chewing- biting Pathway interference  metabolic outcome ? JA-mediated defences Jasmonic acid (JA) Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Feedback on herbivores ? SA-mediated defences Salicylic acid (SA)

3. Theoretical effects of phytohormones on plant chemistry JA-mediated pathwaySA-mediated pathway JA-specific targets SA-specific targets Interaction targets Common targets - SA targets inhibited by JA JA targets inhibited by SA Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany

4. Theoretical effects of phytohormones on plant chemistry JA-mediated pathwaySA-mediated pathway JA-specific targets SA-specific targets Interaction s Common targets - SA targets inhibited by JA JA targets inhibited by SA Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Analyses of leaf AND phloem exudate metabolites Plantago lanceolata Daily application of phytohormones  Control (0.01% Triton, pH 3.1)  JA (0.5 mM)  SA (0.5 mM)  JA + SA (0.5 mM each)  JA + SA (0.25 mM each)

5. Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany JA (0.5 mM) Effects of phytohormones on leaf / phloem metabolome after 72 h (2 treatments) significance (U test, P < 0.05) Volcano plots fold change ( 2) Leaves: 1,414 features, phloem: 36 metabolites N = 10; Mann-Whitney U-tests

6. Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany JA (0.5 mM) SA (0.5 mM) Phytohormone-specific modulation of metabolic pool sizes Effects of phytohormones on leaf / phloem metabolome after 72 h (2 treatments) Some overlap in responses Leaves: 1,414 features, phloem: 36 metabolites N = 10; Mann-Whitney U-tests

7. Effects of phytohormones on leaf / phloem metabolome after 72 h (2 treatments) Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany JA + SA ( 0.25 mM) JA + SA (0.5 mM) Changes in metabolite pool sizes depend on applied conc. Leaves: 1,414 features, phloem: 36 metabolites N = 10; Mann-Whitney U-tests No effects on typical targets (iridoid glycosides)

8. Effects of phytohormones on leaf / phloem metabolome after 72 h (2 treatments with 0.5 mM concentrations) Increased Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany

9. Effects of phytohormones on leaf / phloem metabolome after 72 h (2 treatments with 0.5 mM concentrations) Increased Decreased Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Induction pattern fits to theoretical framework In phloem exudates mainly down-regulation -

10. Effects of phytohormones on leaf / phloem metabolome after 72 h (2 treatments with 0.5 mM concentrations) Down-regulated Effects on chewing / sucking herbivores Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Daily application of phytohormones 0.5 mM (150 pmol)

11. Heliothis virescens Effects of phytohormone-treatments on survival of chewing herbivore Kaplan-Meier, log-rank tests + Bonferroni, N = 10 Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany X - censoring Negative impact of both JA and SA Attenuation of negative effects when hormones applied simultaneously

12. Myzus persicae Negative impact of JA and SA on herbivores of both feeding types Intermediate survival on plants treated with both phytohormones Effects of phytohormone-treatments on survival of sucking herbivore Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany X - censoring Kaplan-Meier, log-rank tests + Bonferroni, N = 10

13. Summary Responses of Plantago lanceolata are highly treatment-specific Complementary, synergistic and antagonistic phytohormone effects fine-tune the plant metabolome - Partial attenuation of effects when hormones applied together → strong interactions between both phytohormone pathways Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Herbivores of both feeding types negatively affected by both phytohormones when applied individually → complementary effects of the hormones Schweiger et al. (2014) Plant Cell Environ 37: 1574-1585

14. Thanks Chemical Ecology group, Bielefeld University Karsten Niehaus, CeBiTec, Bielefeld University

15. Effects of phytohormone-treatments on population dynamics of aphids Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Start: N = 10 x 10 adults negative impact of JA and SA on population growth

16. Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Start: N = 10 x 10 adults Effects of phytohormone-treatments on population dynamics of aphids partial attenuation of negative effects when hormones applied simultaneously

17. → No induction of iridoid glycosides Schweiger, Heise, Persicke, Müller - Chemical Ecology - Bielefeld University, Germany Kruskal-Wallis test, X² = 8.254, df = 4, P = 0.935, N = 10 Effects of phytohormones on target compounds after 72 h (2 treatments)

18. JA/SA pathway crosstalk Adapted from Pieterse et al. (2009) Nat Chem Biol