1. Going home…

2. Three out of many good reasons to go home... Max-Planck Institute for Chemical Ecology Jena „Coevolution of plants and insects“ • Get feedback • Learn glucosinolate analysis • Sow some Pachycladon seeds Poster at the Annual Meeting of the International Society of Chemical Ecology 10th Anniversary of the MPICE, 1997-2007 „ The raison d‘etre of plant secondary substances“ - Fraenkel (1959), Science • Ask for a more money...

3. Some broccoli for lunch? – Research at the Jena MPICE Glucosinolate diversity – The mustard oil bomb of brassicaceous plants Specialist insects disarm the mustard oil bomb: • Pieris rapae (cabbage white butterfly) makes NSP • Plutella xylostella (diamondback moth) desulfates glucosinolates rendering them invisible to myrosinase + desulfatase (P. xylostella) +ESP +NSP (P. rapae)

4. Sampling – 3 Populations per Species P. fastigiata P. enysii F1 1700m E1 1640m E2 1900m F2 1200m E3 1780m F3 1400m

5. How many chemotypes? • Distribution across species & sites? Glucosinolates - Within & Between Species Variation 3MSOP 4MSOB Allyl 5MSOP 3Butenyl 6MSOH 3MTP 7MSOH 5Pentenyl 4MTB 8MSOO 4MOI3M P. enysii P. fastigiata

6. Glucosinolates – Clustering Identifies 5 Chemotypes Distribution (Site, # Individuals) Profile # Found in... 1 P. enysii • 3 chemotypes in P. enysii, 2 in P. fastigiata,no overlap! 2 P. fastigiata • Each site dominated by one chemotype, except F3 3 P. enysii 4 P. enysii • Local adaptation or drift? 5 P. fastigiata

7. 4 Arabidopsis ecoypes to play with: Ler, Col, Ws, Cvi WS Cvi WS Cvi Ler Col Ler Col

8. Predicting genotypes from phenotypes? MAM1 C3 gls C4 gls MT gls Alkenyl glucosinolate Hydroxyalkenyl glucosinolate AL gls AOP2 MS gls HAL gls AOP3

9. Predicting genotypes from phenotypes? MAM1+AOP2+ Cvi WS MAM1- AOP- MAM1+ AOP- Col MAM1- AOP3+ Ler

10. GLS loci likely to segregate in P. enysii and P. fastigiata Pachycladon chemotypes Arabidopsis ecotypes Segregating loci MAM1+ AOP2+ E1 Cvi MAM1- AOP- F1/3 Ws MAM1+ AOP2+ E3 Cvi MAM1- AOP2+ E2 F2/3 n/a

11. Gene Regulation Process Predicted Biochemical Profiles Quercetin & sinapates up in P. enysii TT7, FAH1 up in P. enysii Response to UVB 1. Response to Herbivory AOP2, MAM1 up in P. enysii Glucosinolate biosynthesis C4 & Alkenyl glucosinolates up in P. enysii 2. ESP up in P. enysii ESM1 up in P. fastigiata Glucosinolate hydrolysis P. enysii makes nitriles P. fastigiata makes isothiocyanates 3. Do TT7, FAH1, ESP, ESM1 segregate in P. enysii vs P. fastigiata? Flavonoid analysis at Lincoln University (field samples) Analysis of glucosinolate hydrolysis products at MPICE (greenhouse samples)

12. Ideas for year 2 Comparative transcriptomics, proteomics, metabolomics across Pachycladon radiation Transcriptomics: Microarrays (HortResearch) vs. Solexa (AWC) Proteomics: Paul Haynes (Maquarie University) Metabolomics: GLS and GLS hydrolysis products (Jena) Flavonoids (Lincoln) (super-network by S. Joly, unpublished data) Candidate gene studies (expression and allele frequencies studies) Ecological significance of GLS production? (id herbivores, feeding trials)

13. Premliminary GLS survey across Pachycladon radiation P. wallii P. exilis P. fastigiata P. novae-zealandia P. cheesemanii P. enysii (super-network by S. Joly, unpublished data) GLS patterns don’t follow phylogeny

14. Workshop on multivariate analysis of –omics data • Principal Components Analysis (Ordination) • Multidimensional Scaling (Ordination) • Hierachical Clustering (Classification) • Linear Discriminant Analysis (Classification) (implemented in R) Examples • Volatile bouquets of Medicago leaves challenged with different elicitors • Glucosinolate profiles across Pachycladon populations

15. Going home…