A. M. Yáñez-Serrano, L. Fasbender, J. Kreuzwieser, D. Dubbert, S. Haberstroh, R. Lobo-do-Vale, M. C. Caldeira, C. Werner

Diterpene and other isoprenoid emissions by Mediterranean shrubs 1 Photo by C. Werner Isoprenoids • Isoprene Biosynthesis Number C Volatility - Isoprenoid metabolic pathway • Monoterpenes - MEP (plastids) - MVA (cytosol) • Sesquiterpenes Roles - Oxidation protection at cell level • Diterpenes - Signalling compounds - Atmospheric impact • Sesterterpenes - Reactivity - SOA formation • Triterpenes • Tetraterpenes

• Polyterpenes 2 Choice of family: Cistaceae shrubs

Mediterranean region • High temperature and radiation • favor BVOC emissions

• Climate change preditions • Increased abiotic stresses • Shrub expansion (Caldeira et al., 2015)

• Strong influence in atmospheric chemistry (O3 formation)

Expansion and contraction of Mediterranean biome Klausmeyer and Shaw, 2009 PLoSONE 3 halimifolium Cistus ladanifer Objective

To investigate the isoprenoid emissions of two characteristic Mediterranean shrub

• BVOC emission measurements (controlled and natural conditions) • Plant material extraction Information on biosynthesis of the 13 • C-pyruvate labelling via transpiration stream emitted compounds

4 Methodological set up Controlled conditions: Natural conditions: Uni Freiburg Alentejo, Portugal Halimium halimifolium Cistus ladanifer PTR-TOF-MS and GC-MS GC-MS

5 We found the diterpene Kaurene !!!

PTR-TOF-MS GC-MS

273.2 274.2

6 Diurnal courses of kaurene for H. Halimfolium

7 Diurnal courses of isoprene, monoterpenes and sesquiterpenes for H. Halimfolium

Isoprene Monoterpenes Sesquiterpenes

8 Plant material extraction

Relation between emissions and pools suggests diterpene emissions come from storage pools

9 Labelling experiment

Indication of freshly produced BVOC 13C-pyruvate labelling experiment suggests diterpenes emissions do not come from de novo biosynthesis

10 Natural conditions – temperature dependency

11 Conclusions from this study

• Two Cistaceae species emit considerable amount of diterpenes

• Strong diversity in emissions magnitude

• Diterpene emissions derive from storage pools • Plant extraction • Labelling

• Possible temperature dependency – as seen with the natural conditions data

12 Implications

• High reactivity of kaurene • Towards ozone 1.2x10-17 cm3 molec-1 s-1 Similar to a-pinene (EPI) • Towards OH 7.2x10-13 cm3 molec-1 s-1

• Unknown role on aerosol formation potential

• Missing reactivity

• No consideration of this type of compound in any atmospheric chemistry model

13 Acknowledgement

• We thank Boris Bonn and Efstratios Bourtsoukidis for fruitful discussions. • ERC grant number: #647008

Thank you for listening Any questions? 14 15 Direct comparison GC-MS vs PTR-ToF-MS

16 Comparison

• Diterpenes are emitted by Mediterranean shrub species in similar magnitudes to monoterpeneoids and sesquiterpenoids • H. Halimifloium is not an isoprene emitter consistent with other Cistaceae species Unit This study Matsunaga et al., Jardine et al., Bracho-Nunes et -1 -1 2012 2014 al., 2013 µg gdw h H.Halimifolium Mediterranean C. Ladanifer Japanese ecosystems H.Halimifolium species 0.15 to 5.38 0.01 to 7.1 - - Diterpenes average of 0.53 average of 0.61 Monoterpenoids 0.62 - 0.7 16.18-0.13

Sesquiterpenoids 2.1 - 2.43 0.03-0.63 17