Archaea DLO-like N-fixers Metal-reducers Methanotroph NH 3 + oxidizer Acetogens S-oxidizer High...
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Archaea DLO-like N-fixers Metal-reducers Methanotroph NH 3 + oxidizer Acetogens S-oxidizer S-oxidizer High Density Microarray analysis of prokaryotes in planktonic and High Density Microarray analysis of prokaryotes in planktonic and biofilm gold mine communities. biofilm gold mine communities. Comparison of planktonic and biofilm microbial communities in Comparison of planktonic and biofilm microbial communities in fracture water from the deep subsurface of the Witwatersrand fracture water from the deep subsurface of the Witwatersrand Basin, South Africa. Basin, South Africa. Terry Hazen Terry Hazen 1 1 , , Eoin Brodie Eoin Brodie 1 1 , Fred Brockman , Fred Brockman 2 2 , Duane Moser , Duane Moser 2 2 , Tom Gihring , Tom Gihring 2 2 , David Culley , David Culley 2 2 , Li-Hung Lin , Li-Hung Lin 3 3 , , Thomas Pray Thomas Pray 4 4 , Gary Andersen , Gary Andersen 1 1 , Paul Richardson , Paul Richardson 1,5 1,5 , Lisa Pratt , Lisa Pratt 6 6 and Tullis Onstott and Tullis Onstott 4 4 . . 1 Lawrence Berkeley National Lab, 2 Pacific Northwest National Lab, 3 National Taiwan University, 4 Princeton University, 5 Joint Genome Institute, 6 Indiana University Background Background A deep-branching clade of nearly identical Desulfotomaculum-like, or DLO, 16S rDNA sequences (>99% homology) has been identified as the dominant microorganism in fracture water (14 Kyr to 20 Myr old) from multiple Au mines of the Witwatersrand Basin. The DLO is only the dominant form in the planktonic phase of the deepest (2 – 3 km depth), most saline fracture water and has not been reported as part of the community structure in biofilms that occupy deep, open boreholes with flowing fracture water. Here we compare fracture water planktonic communities from a recently drilled borehole in one mine with biofilm samples collected from an existing borehole at another mine in the Witwatersrand Basin. Methods Methods DNA from fissure water and rock biofilm samples collected from 3m, 6m and 9m into a borehole with fracture water ~ 3 Myr old, was amplified, fragmented and hybridized to our high density 16S microarrays containing 500,000 probes capable of detecting 9,381 prokaryotic OTUs. Clone libraries of 16S rDNA were also generated in order to validate array results. Scanning electron microscopy was used to visualize biofilm architecture, while XRD and EDX-S were used to confirm mineral identities. Results Results Planktonic community composition Planktonic community composition Biofilm community composition Biofilm community composition = detected in clone library and array = detected by array only = present in planktonic also = only present in biofilm FeS 2 framboid BaSO 3 crystals Possibly FeS 2 ZnS mineral coating Biofilm interior with various mineral morphologies Biofilm cross section showing dense exopolysaccharide matrix Conclusions Conclusions Biofilm microbial communities in open boreholes are considerably more diverse than planktonic communities present in fracture water and appear to contain many of the functional groups expected given the geochemistry of an ecosystem where highly reduced, metal, sulfide, H 2 and CH 4 rich water encounter oxygenated mine air. Acknowledgements Acknowledgements Planktonic Biofilm Temperature ( o C) 60 31 Eh (mV) -330 to -260 -250 pH 9.2 to 9.3 7.4 Depth sampled (km below surface) 2.83 1.47 CH 4 9 to 17 29 SO 4 2- /S 2- 0.5 to 1.9/1.1 to 1.4 0.55/2.7x10 3 Fe < 2 to 7x10 -4 1.3x10 3 Zn < 2x10 -4 2.8x10 3 Ba 0.2 to 2.5x10 -3 0.55 δ 13 C-CH 4 (‰VPDB) δ 2 H-CH 4 (‰VSMOW) -31.6 to -33.2 -364 to -390 mM/L -56.9 -175 mM/kg Harmony Gold Mine, Inc. for providing access to the boreholes at Evander Mine. Evander geologists Colin Ralston and Pete Roberts. This research was supported by grant EAR-9978267 from the National Science Foundation LExEn program to T.C. Onstott. Physical and chemical characteristics of planktonic and biofilm samples Physical and chemical characteristics of planktonic and biofilm samples Contacts: Contacts: [email protected] – [email protected] – [email protected] Low intensity High intensity
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Transcript of Archaea DLO-like N-fixers Metal-reducers Methanotroph NH 3 + oxidizer Acetogens S-oxidizer High...
Archaea
DLO-like
N-fixers
Metal-reducers
MethanotrophNH3
+ oxidizer
Acetogens
S-oxidizer
S-oxidizer
High Density Microarray analysis of prokaryotes in planktonic and biofilm gold High Density Microarray analysis of prokaryotes in planktonic and biofilm gold mine communities.mine communities.
Comparison of planktonic and biofilm microbial communities in fracture Comparison of planktonic and biofilm microbial communities in fracture water from the deep subsurface of the Witwatersrand Basin, South Africa. water from the deep subsurface of the Witwatersrand Basin, South Africa.
Terry HazenTerry Hazen11, , Eoin BrodieEoin Brodie11, Fred Brockman, Fred Brockman22, Duane Moser, Duane Moser22, Tom Gihring, Tom Gihring22, David Culley, David Culley22, Li-Hung Lin, Li-Hung Lin33, Thomas Pray, Thomas Pray44, , Gary AndersenGary Andersen11, Paul Richardson, Paul Richardson1,51,5, Lisa Pratt, Lisa Pratt66 and Tullis Onstott and Tullis Onstott44..
1Lawrence Berkeley National Lab, 2Pacific Northwest National Lab, 3National Taiwan University, 4Princeton University, 5Joint Genome Institute, 6Indiana University
BackgroundBackground
A deep-branching clade of nearly identical Desulfotomaculum-like, or DLO, 16S rDNA sequences (>99% homology) has been identified as the dominant microorganism in fracture water (14 Kyr to 20 Myr old) from multiple Au mines of the Witwatersrand Basin. The DLO is only the dominant form in the planktonic phase of the deepest (2 – 3 km depth), most saline fracture water and has not been reported as part of the community structure in biofilms that occupy deep, open boreholes with flowing fracture water. Here we compare fracture water planktonic communities from a recently drilled borehole in one mine with biofilm samples collected from an existing borehole at another mine in the Witwatersrand Basin.
MethodsMethods
DNA from fissure water and rock biofilm samples collected from 3m, 6m and 9m into a borehole with fracture water ~ 3 Myr old, was amplified, fragmented and hybridized to our high density 16S microarrays containing 500,000 probes capable of detecting 9,381 prokaryotic OTUs. Clone libraries of 16S rDNA were also generated in order to validate array results.Scanning electron microscopywas used to visualize biofilmarchitecture, while XRD and EDX-S were used to confirm mineral identities.
ResultsResults
Planktonic community compositionPlanktonic community composition Biofilm community compositionBiofilm community composition= detected in clone library and array= detected by array only
= present in planktonic also= only present in biofilm
FeS2 framboid
BaSO3 crystals
Possibly FeS2
ZnS mineral coating
Biofilm interior with various mineral morphologies
Biofilm cross section showing dense exopolysaccharide matrix
ConclusionsConclusions
Biofilm microbial communities in open boreholes are considerably more diverse than planktonic communities present in fracture water and appear to contain many of the functional groups expected given the geochemistry of an ecosystem where highly reduced, metal,
sulfide, H2 and CH4 rich water encounter oxygenated mine air.
AcknowledgementsAcknowledgements
Planktonic Biofilm
Temperature (oC) 60 31
Eh (mV) -330 to -260 -250
pH 9.2 to 9.3 7.4
Depth sampled (km below surface) 2.83 1.47
CH4 9 to 17 29
SO42-/S2- 0.5 to 1.9/1.1 to 1.4 0.55/2.7x103
Fe < 2 to 7x10-4 1.3x103
Zn < 2x10-4 2.8x103
Ba 0.2 to 2.5x10-3 0.55
δ13C-CH4 (‰VPDB)
δ2H-CH4 (‰VSMOW)
-31.6 to -33.2
-364 to -390
mM/L
-56.9
-175
mM/kg
Harmony Gold Mine, Inc. for providing access to the boreholes at Evander Mine. Evander geologists Colin Ralston and Pete Roberts. This research was supported by grant EAR-9978267 from the National Science Foundation LExEn program to T.C. Onstott.
Physical and chemical characteristics of planktonic and biofilm samplesPhysical and chemical characteristics of planktonic and biofilm samples
Contacts:Contacts: [email protected] – [email protected] – [email protected]
Low intensity High intensity
