abundance and structure of microorganisms related to methane cycling in five european peatlands:...
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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
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
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)