1. Abundance and structure of microorganisms related to methane cycling in five European peatlands: Influence of plant cover and restoration stage (WP1) A. Gattinger et al. Technical University of Munich (at the campus of GSF-Research Center for Environment & Health) Chair of Soil Ecology D- 85764 Neuherberg

2. I. Depth distribution of archaeal (methanogenic) biomass among countries Country x Depth i20:0 I20:1 I40:0 i40:1cyx i40:1cy i40:2cyx i40:2cy Finland France/B France/LScotland Switzerland 0,00 20,00 40,00 60,00 80,00 100,00 PLEL_(nmol/g dm) 329 Depth 3 4 6 8 Finland France/B France/LScotland Switzerland Pooled for „country“

3. I. Depth distribution of methanotrophic biomass (Type I and II) among countries Finland France/B France/L ScotlandSwitzerland PLFA (nmol/g dry matter) Country x Depth 0,00 20,00 40,00 60,00 80,00 100,00 120,00 140,00 Depth 2 3 4 6 8 Finland France/B France/LScotland Switzerland Country 0,00 5,00 10,00 15,00 Type I Type II Finland France/B France/LScotland Switzerland 15,00 Pooled for „country“ Country x Depth

4. II. Methanogens to methanotrophs along gradients Finnland Jura sites

5. …the presented data should be combined with CH4/CO2 flux data for making system-related studies. ………………………………………………………………………………….. PLFA analyses completed (WP1) apart from specific biomarkers, data of other PLFA is avavailable (saturated, mono-unsaturated and polyunsaturated fatty acids) for calculating total biomasses of eg. Bacteria, fungi, etc.

6. Labelled litter 13 C - 15 N 15 N mineralization towards microbes Microbial communities : 13 C PLFA analysis 13 C & 15 N in microbial biomass towards peat 13 C & 15 N (K 2 SO 4 extract without fumigation) Peat column WP3: Microbial transformations of plant litter (TUM-BO, ECOBIO, EPFL/WSL) (TUM-BO, ECOBIO, EPFL/WSL)

7. What do we want to know? 1.Carbon transformation  How much plant C is consumed by the microbial biomass?  How much plant C is used by Bacteria, Archaea, Eukarya and when?  How much plant C is „somehow“ stabilised? 2. N transformation  How much plant N is consumed by the microbial biomass?  How much plant N is mineralised?  How much plant N is somehow stabilised? Labelled litter 13 C - 15 N 15 N mineralization towards microbes Microbial communities : 13 C PLFA analysis 13 C & 15 N in microbial biomass towards peat 13 C & 15 N (K 2 SO 4 extract without fumigation) Peat column

8. Simultaneous identification and quantification of PLFA/PLEL from environmental samples and their corresponding 12 C/ 13 C ratios by GC/MS-c-IRMS MS (DSQ) IRMS (DeltaPlus Advantage ) 20% of the analyte 80% of the analyte Agilent MSD

9. SATFAs (Bacteria) PUFAs (Eukarya)

10. Bacteria: Gram-positive (i15:0) ControlSphagnumEriophorum -30 -20 -10 0 10 20 30 40 50 60 δ 13C (‰) 123123 sampling date (= 15d, 60d, 150d) 41 184 123 81 Horizon 012

11. Bacteria: Gram-positive (a15:0) ControlSphagnumEriophorum -30 -20 -10 0 10 20 30 40 50 60 δ 13C (‰) 123123 sampling date 123 Horizon 012 145 179 186 20

12. Bacteria: Gram-negative (cy17:0) ControlSphagnumEriophorum δ 13C (‰) 123123 sampling date 123 Horizon 012 83 106 56 184 21 175 23 -40 -30 -20 -10 0 10 20 30 40 50 60

13. Bacteria: Gram-negative (cy19:0) ControlSphagnumEriophorum δ 13C (‰) 123123 sampling date 123 Horizon 012 -40 -30 -20 -10 0 10 20 30 40 50 60 107 184

14. Archaea: Euryarchaeota (i20:0) ControlSphagnumEriophorum δ 13C (‰) 123123 sampling date 123 Horizon 012 108 101 186 -40 -30 -20 -10 0 10 20 30 40 50 60 70 80

15. Eukarya: Fungi (18:2d9,12) ControlSphagnumEriophorum δ 13C (‰) 123123 sampling date 123 Horizon 012 -40 -30 -20 -10 0 10 20 30 40 50 60 70 41 35

16. Eukarya: Protozoa (20:4d5,8,11,14) ControlSphagnumEriophorum δ 13C (‰) 123123 sampling date 123 Horizon 012 47 161 142 33 -40 -30 -20 -10 0 10 20 30 40 12 9

17. What needs to be done? 1.Carbon transformations  LC/IRMS of prepared microbial biomass C (CFE) extracts  Calculation of group-specific microbial plant C utilisation  Mass balance for the „whole“ system 2. Nitrogen transformations  EA-IRMS of prepared microbial biomass N (CFE) extracts  EA-IRMS of mineral N extracts (?)  Calculation of microbial plant N utilisation  Mass balance for the whole system

18. LC IsoLink

19. Chromatogramm Standard

20. Reproduzierbarkeit des LC/IRMS-Signals δ 13 C [‰ PDB] n = 3 + SE Neue Methode ist:  reproduzierbar stabil  sensitiv  effizient (ca. 100 Proben/Tag)