0 Microbial Gene Research & Resources Facility (MGRRF) Institute for Molecular & Cellular Therapies (IMCT) Biomolecular & Biomedical Sciences (BBS) New.

1Microbial Gene Research & Resources Facility (MGRRF)Institute for Molecular & Cellular Therapies (IMCT)Biomolecular & Biomedical Sciences (BBS)

New Lorne Bore (RN 17263). Depth 1613 m, Temp: 89 oC, pH 8.5); Flow rate: 7.56 L/sec; Chemistry: typical Na-HCO3-Cl

Brisbane

Sampling at bore source

Sampling below bore source

2Microbial Gene Research & Resources Facility (MGRRF)Institute for Molecular & Cellular Therapies (IMCT)Biomolecular & Biomedical Sciences (BBS)

The New Lorne Bore (RN 17263) runoff channel

Temperature specific microbial colonisation occurs in the New Lorne bore run off channel in a temp gradient (88 oC to ambient)

Sampling microbial

mats

3Microbial Gene Research & Resources Facility (MGRRF)Institute for Molecular & Cellular Therapies (IMCT)Biomolecular & Biomedical Sciences (BBS)

Grey Filaments at 75ºC Red Filaments at 66ºC

Brown Filaments at 52ºCGreen Filaments at 57ºC4 spatially distinct colored microbial mats colonise

New Lorne bore runoff channel at different temperatures

5 Samples (4 mats + source sample)

HMW DNA extracted

16S rRNA genes cloned

400 clones studied to determine phylogroups

Representative of each phylogroup sequenced

Phylogenetic Reconstruction

Dic

tyog

lom

us

The

rmod

esu

lfob

acte

ria

Bacteroides

The

rmom

icro

bia

Aq

uif

ex

Chr

ysio

gene

tes

The

rmot

oga

Th

erm

ales

Firm

icut

esChlamydia

Def

erri

bact

er

Nit

rosp

ira

Cya

noba

cter

iaActinobacteria

Verrucomicrobia

Acidobacteria

PlanctomycetesFibrobacter

Spirochetes FusiformsPro

teob

acte

ria

Microbial diversity of the New Lorne bore & the 4 runoff channel microbial mats (10 / 21 kingdoms)

1 23 4

5

6

7

8

9

10

6Microbial Gene Research & Resources Facility (MGRRF)Institute for Molecular & Cellular Therapies (IMCT)Biomolecular & Biomedical Sciences (BBS)

Grey Red Green Brown(75ºC) (66ºC) (57ºC) (52ºC)

Bacillus þ

Hydrogenobacter þ þ

Cytophagales þ þ

Clostridia þ þ

Thermus þ þ þ

Meiothermus þ þ þ

Planctomycetes þ þ þ

Chloroflexus þ þ þ

Cyanobacteria þ þ

Acidobacteri þ þa-Proteobacteria þ þ

b-Proteobacteria þ þ

g-Proteobacteria þ þ

d-Proteobacteria þNitrospira þSporomusa þ

Environental Clones þ

Prosthecobacter þ

Group-genus-species distribution

The microbial diversity profile of the 4 microbial mats

Phylum Genus / species Source (89 ºC)

Grey(75 ºC)

Red(66 ºC)

Green(57 ºC)

Brown(52 ºC)

AquificaeH. subterranea 100 6 3

DeinococciThermus T. scotoductus 62 4

T. igniterrae 12 3T. oshimai 1

Meiothermus M. ruber 47 4M. cerberus 2 34

Chloroflexi C. auranticus 4 10 3Nitrospira N. muscoviensis 4Verrucomicrobia 1Cyanobacteria F. musicola 36 1Planctomycetes 1 2 5Acidobacteria 2 8Bacteroidetes 3 6

Proteobacteria

a- Proteobacteria 1 20

b- Proteobacteria 13 2

d- Proteobacteria 2

Firmicutes 8 8 1

Novel,Unaffiliated??

9 24 21 21

Quantitative diversity based on clones (in %)