Abundance and structure of microorganisms related to methane cycling in five European peatlands: Influence of plant cover and restoration stage (WP1) A.

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 EcologyD- 85764 Neuherberg

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/BFrance/L ScotlandSwitzerland

0,00

20,00

40,00

60,00

80,00

100,00

PL

EL

_(n

mo

l/g d

m)

329

Depth3

4

6

8

Finland France/BFrance/L ScotlandSwitzerland

Pooled for „country“

I. Depth distribution of methanotrophic biomass (Type I and II) among countries

Finland France/BFrance/L ScotlandSwitzerland

PL

FA

(n

mo

l/g d

ry m

atte

r)

Country x Depth

0,00

20,00

40,00

60,00

80,00

100,00

120,00

140,00 Depth2

3

4

6

8

Finland France/BFrance/L ScotlandSwitzerland

Country

0,00

5,00

10,00

15,00

Type I

Type II

Finland France/BFrance/L ScotlandSwitzerland

15,00

Pooled for „country“Country x Depth

II. Methanogens to methanotrophs along gradients

0 = bare peat 1 = Evag_dry 2 = Evag_wet 3 = Carex 4 = Sphag

Plants

0

100

200

300

400

500

600

700

Met

hano

gens

to m

etha

notr

ophs

FI- E-8-1

FI- A-4-2

FI- A-6-1

FI- C-3-2

FI- D-6-2Finnland

CH France/L

Country

0

20

40

60

80

100

Met

hano

gens

to m

etha

notr

ophs

FR- B-6-1

CH- C-6-2

CH- A-3-1

CH- B-8-1

FR- C-6-2

FR- C-8-2

FR- D-8-2

FR- D-8-1

Plants0 = patchy

5 = early

6 = advanced

7 = intact

Jura sites

…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.

Labelled litter13C - 15N

15N mineralization

towards microbes

Microbial communities :13C PLFA analysis

13C & 15N in microbial biomass towards peat

13C & 15N (K2SO4 extract without fumigation)

Peat column

WP3: Microbial transformations of plant litterWP3: Microbial transformations of plant litter

(TUM-BO, ECOBIO, EPFL/WSL)(TUM-BO, ECOBIO, EPFL/WSL)

What do we want to know?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 litter13C - 15N

15N mineralization

towards microbes

Microbial communities :13C PLFA analysis

13C & 15N in microbial biomass towards peat

13C & 15N (K2SO4 extract without fumigation)

Peat column

Simultaneous identification and quantification of PLFA/PLEL from environmental samples and their corresponding 12C/13C ratios by

GC/MS-c-IRMS

MS(DSQ)

IRMS(DeltaPlusAdvantage)

20% of the analyte

80% of the analyte

AgilentMSD

SATFAs (Bacteria)

PUFAs (Eukarya)

Bacteria: Gram-positive (i15:0)

Control Sphagnum Eriophorum

-30

-20

-10

0

10

20

30

40

50

60

δ

13C

(‰

)

1 2 3 1 2 3

sampling date (= 15d, 60d, 150d)

41 184

1 2 3

81

Horizon

0 1 2

Bacteria: Gram-positive (a15:0)

Control Sphagnum Eriophorum

-30

-20

-10

0

10

20

30

40

50

60

δ

13C

(‰

)

1 2 3 1 2 3

sampling date

1 2 3

Horizon

0 1 2

145

179

18620

Bacteria: Gram-negative (cy17:0)

Control Sphagnum Eriophorum

δ

13C

(‰

)

1 2 3 1 2 3

sampling date

1 2 3

Horizon

0 1 2

83 10656184

21

17523

-40

-30

-20

-10

0

10

20

30

40

50

60

Bacteria: Gram-negative (cy19:0)

Control Sphagnum Eriophorum

δ

13C

(‰

)

1 2 3 1 2 3

sampling date

1 2 3

Horizon

0 1 2

-40

-30

-20

-10

0

10

20

30

40

50

60

107 184

Archaea: Euryarchaeota (i20:0)

Control Sphagnum Eriophorum

δ

13C

(‰

)

1 2 3 1 2 3

sampling date

1 2 3

Horizon

0 1 2

108

101

186

-40

-30

-20

-10

0

10

20

30

40

50

60

70

80

Eukarya: Fungi (18:2d9,12)

Control Sphagnum Eriophorum

δ

13C

(‰

)

1 2 3 1 2 3

sampling date

1 2 3

Horizon

0 1 2

-40

-30

-20

-10

0

10

20

30

40

50

60

70

41

35

Eukarya: Protozoa (20:4d5,8,11,14)

Control Sphagnum Eriophorum

δ

13C

(‰

)

1 2 3 1 2 3

sampling date

1 2 3

Horizon

0 1 2

47

161

142

33

-40

-30

-20

-10

0

10

20

30

40

12

9

What needs to be done?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

LC IsoLink

Chromatogramm

Standard

Reproduzierbarkeit des LC/IRMS-Signals

δ13C

[‰

PD

B]

Zeit [h]

0 10 20 30 40 50 60-27.10-27.05-27.00-26.95-26.90-26.85-26.80-26.75-26.70-26.65

20 mg Benzoesäure-C l-1 in 0,5 M K2SO4

Zeit [h]

0 10 20 30 40 50 60-27.10-27.05-27.00-26.95-26.90-26.85-26.80-26.75-26.70-26.65

20 mg Benzoesäure-C l-1 in 0,5 M K2SO4

10 mg Benzoesäure-C l-1 in 0,01 M CaCl2

Zeit [h]

0 10 20 30 40 50 60-27.10-27.05-27.00-26.95-26.90-26.85-26.80-26.75-26.70-26.65

20 mg Benzoesäure-C l-1 in 0,5 M K2SO4

10 mg Benzoesäure-C l-1 in 0,01 M CaCl2

2 mg Benzoesäure-C l-1 in 0,01 M CaCl2

n = 3 + SE

Neue Methode ist:

reproduzierbar stabil

sensitiv

effizient (ca. 100 Proben/Tag)