Transcript Andreas Gattinger - The Macaulay Land Use Research Institute

Summary of the current status of the work
of TUM-BO
Andreas Gattinger, Matthias Weiss, Michael Schloter
GSF-Research Center for Environment and Health
Technical University of Munich
Institute of Soil Ecology
D- 85764 Neuherberg
Summary of the current status of the work of
TUM-BO
1. Adaptation of protocols for lipid biomarker analyses to peat
(bog) samples
2. Adaptation of protocols for DNA analyses to peat (bog)
samples
3. Growing of keystone plant species in the greenhouse for plant
litter labelling
4. Installation of the new analytical device GC/MS-C-IRMS for
compound specific isotope analysis
1. Adaptation of protocols for lipid biomarker
analyses to peat (bog) samples
Determination of microbial communites
•Problem: less than 5% of the estimated total soil microflora can be cultivated
•Solution: cultivation-independent approaches enable the characterization of
microbial communiteis in situ:
genotypic approach:
(example archaeal 16S rDNA,
separation with SSCP )
phenotypic approach:
(side chain analysis of polar membrane lipids)
1
PLEL derived hydrocarbons
8
6
3
10
5
9
12
11
2
4
0
7
13
14 15
The biosphere of soil
methanogens
Archaeal kingdoms (Reiche):
- Euryarchaeota
- Crenarchaeota
- (Korarchaeota)
- (Nanoarchaeota)
(mod. Version, M. Schmid, GSF)
Side chain analysis of polar membrane lipids of Bacteria
Eucarya and Archaea in soil
O
O
P
Bacteria & Eucarya
NH3+
O
O
CH2
esterlinked fatty acids (PLFA):
- saturated (SATFA)
- monounsaturated (MUFA)
- polyunsaturated (PUFA)
- hydroxysubstituted (PLOH)
O
C
O
H
O
O
C
O
O
CH2
H
O
O
P
O
O
NH3+
Archaea
etherlinked isoprenoids (PLEL):
- saturated short chain: i20:0
- saturated long chain: i40:0
- cyclic long chain: i40:0-cy
- unsaturated short chain: i20:1
OH
OH
NH
O
Bacteria & Eucarya
non-esterlinked fatty acids (NEL-PLFA):
- hydroxysubstituted (UNOH)
- unsubstituted fatty acids (UNSFA)
Extended phospholipid analysis
Sample
Bligh-Dyer-Extraction
Lipids
SI-column
Neutral lipids
Phospholipids
Glycolipids
Archaea
Bacteria & Eucarya
Ether lipids (PLEL)
Fatty acids (PLFA)
Mild alkaline methanolysis
Acidic hydrolysis
NH2-column
Hydroxy-substituted
(EL-PLFA)
Non-substituted
(EL-PLFA)
Unsaponifiable lipids
Acidic hydrolysis
NH2-column
SCX-column
Alkyl iodides
Reduktive dehalogenation
(NEL-PLFA)
SATFA
MUFA
+
DMDS
PUFA
PLOH
+
TMSI
GC-MS
Bodenökologie
UNSFA
UNOH
+
TMSI
Isoprenoid hydrocarbons
GC-MS
Microbial biomarker composition in a Scottish “test” peat core
SATFA (mainly bacteria)
MUFA (mainly aerobic bacteria)
PUFA (eukaryotes: fungi, protozoa)
PLOH (mainly Gram-negative bacteria)
UNOH (anaerobic bacteria and some fungi)
UNSFA (mainly anaerobic bacteria)
PLEL (archaea)
percentage of biomarker fraction
Soil depth [cm]
8
63
0-6
6-12 10 48 8
63
18-24
69
0
1
13
50
13 2
179
1
3
10 1
175
5
14 1
100
156
191
2
12-18
24-30
Number
200
183
300
400
Lipid biomarker [nMol g-1 dry matter]
500
Lipid biomarker ratios in a Scottish “test” peat core
Bacteria : Fungi
MOB : Archaea
Soil depth [cm]
0-6
6-12
2
12-18
63
18-24
69
13 2
10 1
0
1
3
5
14 1
24-30
1
2
3
20
25
Biomarker ratio [mol mol-1]
30
35
MOB and archaeal biomarker in a Scottish “test” peat core
MOB I
MOB II
Archaea
Soil depth [cm]
0-6
6-12
12-18
18-24
24-30
0
2
4
6
8
-1
10
Lipid biomarker [nmol g dry matter]
12
14
2. Adaptation of protocols for DNA analyses to
peat (bog) samples
together with Rebekka Artz, Steve Chapman/MLURI
3. Growing of keystone plant species in the
greenhouse for plant litter labelling
Plants were taken by A. Siegenthaler et al. at the French/Suisse
site:
4 plant species: 2 mosses (Sphagnum, Polytrichum), 2 grasses
(Eriophorum vaginatum, Eriophorum angustifolium) were
grown at the GSF greenhouse for estimating growth conditions
and biomass production
climatic conditions:
4. Installation of the new analytical device GC/MS-CIRMS for compound specific isotope analysis
Simultaneous identification and quantification of PLFA/PLEL from
environmental samples and their corresponding 12C/13C ratios by
GC/MS-C-IRMS
20% of the analyte
MS
(DSQ)
IRMS
(DeltaPlusAdvantage)
80% of the analyte
delta 13C values in PLFA/PLEL from test core samples varied
between -26 and -32 %0 (natural abundance):
due to dilution effects with the peat soil and 1-2%
measurement inaccuracy, an enrichment with 13C in the plant
litter of at least 70 %0 PDB is needed:
Which quality of CO2 gas for plant labelling?