summary of the current status of the work of tum-bo
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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. - PowerPoint PPT PresentationTRANSCRIPT
Summary of the current status of the work of TUM-BO
Andreas Gattinger, Matthias Weiss, Michael Schloter
GSF-Research Center for Environment and HealthTechnical University of MunichInstitute of Soil EcologyD- 85764 Neuherberg
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
Summary of the current status of the work of TUM-BO
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:
0
1
2
3
4 7
5
6
8
910
11
12
1314 15
PLEL derived hydrocarbons
genotypic approach:(example archaeal 16S rDNA,
separation with SSCP )
phenotypic approach:(side chain analysis of polar membrane lipids)
methanogens
The biosphere of soil
(mod. Version, M. Schmid, GSF)
Archaeal kingdoms (Reiche):Archaeal kingdoms (Reiche):- Euryarchaeota- Euryarchaeota- Crenarchaeota- Crenarchaeota- - (Korarchaeota)(Korarchaeota) - - (Nanoarchaeota)(Nanoarchaeota)
HO
O
CH2
C
O
C
O
O P O
O
O
NH3+
H O
CH2
O
O P O
O
O
NH3+ ArchaeaArchaea
etherlinked isoprenoids (PLEL):- saturated short chain: i20:0- saturated long chain: i40:0- cyclic long chain: i40:0-cy- unsaturated short chain: i20:1
Side chain analysis of polar membrane lipids of BacteriaEucarya and Archaea in soil
Bacteria & Bacteria & EucaryaEucaryaesterlinked fatty acids (PLFA):- saturated (SATFA)- monounsaturated (MUFA)- polyunsaturated (PUFA)- hydroxysubstituted (PLOH)
O
OH OH
NH
Bacteria & Bacteria & EucaryaEucaryanon-esterlinked fatty acids (NEL-PLFA):- hydroxysubstituted (UNOH)- unsubstituted fatty acids (UNSFA)
Extended phospholipid analysis
xBodenökologie
Sam ple
G C-M S
Neutral lipids Glyco lipids
B ligh-D yer-E xtrac tion
L ipids
G C-M S
NH -column2
G C-M S
Ether lipids (PLEL)
Hydroxy--
(EL-PLFA)substituted
SATFA MUFA+DMDS
PUFA PLOH+TMSI
UNSFA UNOH+TMSI
Reduktive dehalogenation
A lkyl iodides
Isoprenoid hydrocarbons
SCX-column NH -column2
SI-column
Phospholipids
Acid ic hydrolysis
Non-substituted (EL-PLFA)
Unsaponifiable lipids
Mild alkaline methanolysis
(NEL-PLFA)
Fatty acids (PLFA)Acid ic hydrolysis
ArchaeaBacteria & Eucarya
Lipid biomarker [nMol g-1 dry matter]
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)
100 200 300 400 5000
0-6
6-12
12-18
18-24
24-30
So
il d
ep
th [
cm
]
63
63
69
50
10
8
48
13
10
14
13
8
2
1
1
1
1
2
3
5
156
191
179
175
183
Numberpercentage of biomarker fraction
Microbial biomarker composition in a Scottish “test” peat core
63
69
13
10
14
2
1
1
1
2
3
5
Biomarker ratio [mol mol-1]
0 1 2 3 20 25 30 35
Bacteria : FungiMOB : Archaea
So
il d
ep
th [
cm
]
0-6
6-12
12-18
18-24
24-30
Lipid biomarker ratios in a Scottish “test” peat core
MOB IMOB IIArchaea
2 4 6 8 10 12 140
So
il d
epth
[cm
]
0-6
6-12
12-18
18-24
24-30
Lipid biomarker [nmol g-1dry matter]
MOB and archaeal biomarker in a Scottish “test” peat core
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-C-IRMS 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
MS(DSQ)
IRMS(DeltaPlusAdvantage)
20% of the analyte
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?