1. www.TrophinOak.de Endogenous rhythmic growth, a trait suitable for the study of interplays between multitrophic interactions and tree development Herrmann S. 1, Grams T.E.E. 2, Tarkka M.T. 1, Angay O. 2, Bacht M. 3, Bönn M. 1, Feldhahn L. 1, Graf M. 4, Kurth F. 1, Maboreke H. 5, Mailander S. 6, Recht S. 1, Fleischmann F. 2, Ruess L. 5, Schädler M. 1, Scheu S. 4, Schrey S. 6, Buscot F. 1 1.Helmholtz Centre for Environmental Research (Halle, Saale), 2.Technical University Munich, 3.University of Marburg, 4.University of Göttingen, 5.Humboldt University Berlin, 6.University of Tübingen Herrmann et al. 2016, PPEES 19, 40-48

2. Page 2 TrophinOak platform: DF159, a clone of Q. robur Synthesis of biotic interactions on rhythmic growing trees in Microcosms Herrmann et al. 2016, PPEES 19, 40-48 Microcosms shoots + leaves Lateral roots At 25°C and a 16/8 day/night light regime oaks display endogenous rhythmic growth with alternating flushes in root and shoots Harmer R 1990, New Phytol 115, 23-37

3. SEITE 3 Objectives of TrophinOak Herrmann et al. 2016, PPEES 19, 40-48

4. SEITE 4 mobile labeling chamber Experiment Piloderma und Phytophthora Incubation and labeling with 13 CO 2 Angay et al. (2014) New Phytol. 203:1282-1290 The TrophinOak platform: 13 C-labeling chamber Herrmann et al. 2016, PPEES 19, 40-48

5. SEITE 5 16 normalized Q. robur DF159 cDNA pools 454 pyrosequencing (UFZ) 278,777 trimmed sequences MIRA Conversion to pseudo- Illumina reads 454OakContigDF159 assembly 8 Q. robur DF159 cDNA pools Illumina sequencing 110,276,398 trimmed PE reads Trinity OakContigDF159.1 hybrid assembly 65,712 contigs 1,003 bp mean length Oaks genome resources Merging with INRA Bordeaux RNA Seq library Lesur I et al. (2015) BMC Genomics Oak genome sequencing by INRA Bordeaux/Genoscope (France) DF159 genome re-sequencing by Genoscope Tarkka et al. (2013) New Phytol. 199: 529-540 The TrophinOak platform: Oak Contig library DF159.1

6. Page 6 Stage B Stage C Stage D Stage A Shoot elongation Shoot growth cessation Leaf expansion Swelling bud Root flush RF EM Piloderma croceum Interplay between rhythmic growth & biotic interactions C & N Allocations, RNA Seq Herrmann et al. 2016, PPEES 19, 40-48 Shoot flush SF EM / Interactor X 6 different partners

7. SEITE 7 13 C Excess and 15 N Excess enhanced by P. croceum, but no impact on rhyhmic growth Herrmann et al. (2015) J Exp Bot 66, 7113-7127 Sink Leaf (■) Source Leaf-1 (■) Source Leaf-2 (■) Stem (■) Principal Roots (■) Lateral Roots (■) Endogenous rhythmic growth impacts on C & N allocations during SF (A-B) to RF (C-D) transitions Control P. croceum Rhythmic growth not resource driven Interplays between rhythmic growth, resource shifts and mycorrhizal effects

8. SEITE 8 Angay et al. (2014) New Phytol. 203:1282-1290 Biotic interactions stimulated by resource shifts during ERG SF Pilo RF Pilo SF Pilo RF Pilo SF RF Root infection measured by DNA qPCR 7 D after inoculation Phytophthora q. vs. control Control oaks vs. EM oaks. X Similar patterns with nematodes and mildew Caravaca et al. (2015) Soil Biol. Biochem. 82, 65-73 Mailander et al. unpublished

9. SEITE 9 Piloderma croceum (ectomycorrhiza) Kurth et al. (2015) BMC Genomics 16 ( art. 658) AcH 505 AcH 505 + P. croceum Interplays between growth flushing and biotic interactions differential gene expression Streptomyces sp. AcH 505 (mycorrhization helper bacteria) Poster S3.5 by Tarkka et al.

10. SEITE 10 Stage B Stage C Stage D Stage A Interplay rhythmic growth, mycorrhizal interaction Transition to bud rest Transition to root rest Piloderma croceum (Pi) Control Plants (Co) Herrmann et al. J. Exp Bot (2015) 66, 7113-7127

11. SEITE 11 Differential expression of photoperiod and circadian clock related genes Up-regulated contigs Down-regulated contigs Herrmann et al. J. Exp Bot (2015) Core genes regulated during rhythmic growth

12. SEITE 12 Herrmann et al. 2016, PPEES 19, 40-48 Interplay rhythmic growth # biotic interactions, looking for gene regulation patterns Transcriptome change compared to non-inoculated control plants

13. SEITE 13 Summary 1.Endogenous rhythmic growth impacts on biotic interactions a.Shifts in below- aboveground resource allocations b.Resource shifts impact on virulence of the interactors c.Rhythmic growth not driven by resources but by internal clock processes 2.Endogenous rhythmic growth interplays with biotic interactions at gene regulation level a.Each biotic partner interplays in both shoot and roots at a specific development phase (RS vs. SF) b.No relation to targeted plant part or interaction type (beneficial vs. detrimental) c.ERG a tool to tackle key (core) genes for both development and interactions

14. SEITE 14 Thank you for your attention! www.TrophinOak.de DF159 enjoying freedom The TrophinOak-PhytOmeter platform