laboratory of microbial ecology cees/rug
DESCRIPTION
Work package 3 Analysis of the species diversity, community structure and phylogeny of microorganisms. Laboratory of Microbial Ecology CEES/RUG. Objectives. Evaluate microbial communities in 4 deep sea hypersaline anoxic basins (DHABs) - PowerPoint PPT PresentationTRANSCRIPT
Work package 3Analysis of the species diversity,
community structure and phylogeny of microorganisms.
Laboratory of Microbial Ecology
CEES/RUG
Objectives• Evaluate microbial communities in 4 deep sea
hypersaline anoxic basins (DHABs)
• Evaluate similarities between DHABs and between different positions in one DHAB
• Link previously published geochemical data with these results
Brine interface waterFilter
DNA-isolation
Bacterial 16S rDNA
PCR
Archaeal 16S rDNA
PCR
Cloning PCR-product
Sequencing 700 bases
Sequencing 700 bases
Cloning PCR-product
Bacteria
Brine Interface Seawater
Atalante Bannock Discovery Urania Atalante Bannock Discovery Urania Bannock Discovery
Sequences 90 90 90 90 90 90 90 90 90 90
OTUa 40 41 48 16 28 27 36 25 39 57
Shannon 3.26 3.20 3.51 2.17 2.90 2.72 3.27 2.78 3.42 3.88
Archaea
Brine Interface Seawater
Atalante Bannock Discovery Urania Atalante Bannock Discovery Urania Bannock Discovery
Sequences 90 90 90 90 90 90 90 90 90 90
OTU 11 10 10 5 15 14 15 16 17 12
Shannon 1.60 1.49 1.55 0.61 2.01 1.83 1.77 1.44 2.30 1.87
PCoA of Morisita’s Index for Bacterial OTU-level
17.9
% o
f V
aria
nce
1 9 .9% o f Var ia n ce
C o ord in ate 1
Coo
rdin
ate
2
U IB I
D I
A IB B
A B
D B
D 3 30 0
B 3 00 0
U B
0 .30 .20 .10-0.1-0.2-0.3 0 .60 .50 .4
-0.3
0 .1
0 .2
0 .6
-0.2
0-0.1
0 .4
0 .5
0 .7
0 .3
PCoA of Morisita’s Index for Archaeal OTU-level
U B
18.2
% o
f V
aria
nce
5 1 .5% o f Var ia n ce
C o ord in ate 1
Coo
rdin
ate
2
-0.4 0.40 .30 .20 .10-0.1-0.2-0.3
-0.4
0 .4
0 .8
0 .2
0 .6
-0.2
0
U I
B I
D I
A I B BA B
D B
D 3 30 0
B 3 00 0
Cluster Analysis (UPMGA) of Morisita’s Index for OTU-level
Bacterial Brine+Interface
Archaeal Brine
Archaeal Interface
BB AB DBUB
0.2
0.6
0.4
0.8
1.0
PCoA of Morisita’s Index for Bacterial Phylogenetic Group-level
23.3
% o
f V
aria
nce
4 5 .3% o f Var ia n ce
C o ord in ate 1
Coo
rdin
ate
2
U I
B I
D I
A I
B B
A BD B
D 3 30 0
B 3 00 0
U B
0 .30 .20 .10-0.1-0.2-0.3 0 .50 .4
-0.3
0 .1
0 .2
-0.2
0
-0.1
0 .4
0 .3
-0.4
-0.4
-0.5
0
7.6%
of
Var
ianc
e
8 8 .0% o f Var ia n ce
C o ord in ate 1
Coo
rdin
ate
2
U I
B I
D I
A IB B
A B
D B
D 3 30 0B 3 00 0 U B
0 .30 .20 .10-0.1-0.2-0.3-0.4
0 .2
0 .6
-0.2
0 .4
0 .8
PCoA of Morisita’s Index for Archaeal Phylogenetic Group-level
Principal Component Analysis
• The two seawater samples always group together
• Discovery brine and Discovery Interface always group together
• L’Atalante, Urania and Bannock brines group together except for Bannock Bacterial
• L’Atalante, Urania and Bannock interface group together except for L’Atalante Archaea
• In most cases, Discovery samples group together with other brine samples
Cluster analysis• Discovery is most different from the
other DHABs for the bacterial and archaeal brine and the bacterial interface samples.
• Next to Discovery, Urania is most different from the other DHABs
• Two clusters were observed for the archaeal interface samples
0.0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
40.0
45.0
50.0
delt
a-pr
oteo
Bac
tero
idet
es
epsi
lon
prot
eo
new
/unk
now
n5 ga
mm
a-pr
oteo
KB
1
AB
BB
DB
UB
AI
BI
DI
UI
B3000
D3300
Percentage (%)
Relative Distribution of Phylogenetic Bacterial Groups
0
10
20
30
40
50
60
70
80
90
100
Marine Group I MSBL1 Halobacteriales
AB
BB
DB
UB
AI
BI
DI
UI
B3000
D3300
Percentage (%)
Relative Distribution of Phylogenetic Archaeal Groups
Metabolic Routes
- High number of δ-proteobacteria Sulfate reduction Important
H2, organic matter or anaerobic CH4 oxid.
No archaea related to anaerobic CH4 oxid.
δ-13C CO2 values are not extremely low
H2 and organic matter most important for SO4-reduction
Metabolic Routes
• ε- and γ-proteobacteria were mainly related to sulfide oxidizing bacteria.
• These sequences were most numerous in Urania
• Sulfide oxidation plays an important role in the interface samples
• High number of Bacteroidetes shows that heterotrophic bacteria play important role
Archaea
• Archaea related to Marine group I and a new cluster MSBL1
• At present unknown what the function is of these archaea
• Discovery high number of Halorhabdus utahensis
• This bacterium adapted to high concentrations of Mg2+
• Interface samples different from brine samples. The sharp increase in salinity, sulfide and decrease in O2 are very important factors for the establishment of a microbial community.
• Discovery different. MgCl2 is thus important factor in establishment of microbial community
• Next to Discovery, Urania differs. Sulfide concentration is an important factor as well.
Conclusions
Conclusions• Microbial routes present in the sulfur cycle
have an important function in the DHABs
• Most of the archaeal sequences form a new branch in the archaea tree. This cluster is named MSBL1
• The function of almost all archaea as well for a great number of bacteria is at present unknown
2 A2 B
1 A1 B
3 A
3 B
4 B4 A
33.3
% o
f V
aria
nce
4 5 .5% o f Var ia n ce
Bannock interface samples cruise II
Future Plans• A complete phylogenetic analysis with
GenBank database of the Bannock interface samples
• Study temporal variation by comparing sequences from brines of cruise I, II (and III?)
• Study diversity of several functional genes or specific groups with specific 16S rDNA primers