1. Transcriptome and proteome studies of New Zealand alpine Pachycladon NZPRN 2009 Claudia Voelckel, Mehdi Mirzaei, Michael Reichelt, Luke Luo, Dana Pascovici, Peter B Heenan, Sylvia S Schmidt, Bart Janssen, Paul A Haynes, Peter J Lockhart

2. Q: Adaptive diversification? Q: Mediated by which genes? A: Comparative gene expression profiling in common gardens FA ST EN LA CH EX NZ WA Questions & Approach P. cheesemanii (CH) P. exile (EX) P. novae-zelandiae (NZ) CH EX NZ Pachycladon super net work, S. Joly, unpubl.

3. Lincoln Plant growth Peter Heenan Murray Dawson Auckland Microarray analysis Bart Janssen Luke Luo Silvia Schmidt Jena Glucosinolate analysis Michael Reichelt Palmy Link all data Claudia Voelckel Pete Lockhart Sydney Protein analysis Paul A. Haynes Mehdi Mirzai Dana Pascovici People who helped: almost submitted

4. 1.Protein profiling as a validation of transcript profiling? How much overlap? Specific questions: 3.To what extend will transcript and protein profiling make different predictions regarding downstream physiological differences? 4.Will phylogenetic relationships be reflected in transcript, protein and metabolite profiles? 2.Can we predict physiological differences between CH, EX and NZ and what are they?

5. CH EX+NZEXNZ T37169936 P12460141 EX CH+NZCHNZ T13445946 P109161110 NZ CH+EXCHEX T305221297 P321359211 9601 loci 1489 loci Results 85274151074 TPTP CHEXNZ CH0.520.430.30 EX0.470.450.32 NZ0.400.360.34 T P Overall correlation:Specifically up-regulated genes: T = transcript profiling, P = protein profiling

6. 976129* TPTP EX+NZ 23% 32% CH 18814 142288 TPTP TP CH+NZ CH+EX 18% 4% 36% 3% 2204456 * 1836327 * 131439 * TPTP TP TP NZ EX 20% 56% 13% 30% 40% 6% 20414 * 51251 112479 * TPTP TP TP EX CH 16% 9% 16% <1% 45% 4% EX NZ Specific Genes Found by T AND P

7. Interconversion of carbon dioxide and bicarbonate (carbonic anhydrase) Draught response Serine racemase Vegetative storage proteins 976129* 18814 142288 TPTP TP TP EX+NZ CH+NZ CH+EX 23% 32% 18% 4% 36% 3% CH EX NZ Specific Genes Found by T AND P

8. TEX+NZEXNZ CHAL up C4 up CH+NZCHNZ EX--- CH+EXCHEX NZ--- PEX+NZEXNZ CH-C4 up- CH+NZCHNZ EX--- CH+EXCHEX NZC4 up Predictions from T and P: Glucosinolate Biosynthesis CHEXNZCHEXNZ

9. EX+NZEXNZ CH--- CH+NZCHNZ EXiso? ? CH+EXCHEX NZ??- EX+NZEXNZ CH--- CH+NZCHNZ EX?-- CH+EXCHEX NZnitriles ?? nitriles ?? nitriles ?? Predictions from T and P: Glucosinolate Hydrolysis P. cheesemaniiP. novae-zelandiae

10. TCHEXNZ CH10.910.74 EX10.83 NZ1 PCHEXNZ CH10.750.59 EX10.72 NZ1 == Profiling Patterns Through the Phylogenetic Lens: CH EX NZ ≠ Glucosinolates

11. 1.Protein profiling as a validation of transcript profiling? How much overlap? – overlap small but significant Some Answers: 3.To what extend will transcript and protein profiling make different predictions regarding downstream physiological differences? glucosinolate phenotypes better predicted by protein profiling 4.Will phylogenetic relationships be reflected in transcript, protein and metabolite profiles? – not in metabolite profiles, selection? 2.Can we predict physiological differences between CH, EX and NZ and what are they? – yes (C02 enzymes, draught response, glucosinolate metabolism)

12. Thanks to: YOU! Funding Marsden & Humboldt Foundation New Zealand Landcare: Peter Heenan, Murray Dawson, Kat Trought Plant and Food: Bart Janssen, Luke Luo, Silvia Schmidt AWC: Pete Lockhart, Christine Reinsch, Hanna Daniel Germany MPICE: Michael Reichelt, Jonathan Gershenzon Australia Macquarie University: Mehdi Mirzai, Dana Pascovici, Paul Haynes, Mark Westoby