The 1000 Springs Project
– a bioinventory of New
Zealand’s geothermal
microbial populations
Matthew Stott
21 February, 2017
eResearch NZ 2017
Queenstown
Today’s take-home points
• Microorganisms rule! They:
– are everywhere
– dominant global biomass, diversity,
genetic novelty
– influence all ecosystems (local,
regional, global scales)
• 1000 Springs Project:
– Why?
– How was it set up?
– Outputs?
– Who benefits?
– Future?
Why should we care....
Microorganisms are EVERYWHERE and involved in
practically EVERYTHING you can think of.
Microorganisms rule the Earth!
There are:
~ seven billion (7 x 109) humans on earth
(7,000,000,000)
~ one billion (1 x 109) microorganisms per g of soil
(1,000,000,000)
~ 4-6 x 1030 bacteria/archaea on the earth
(4,000,000,000,000,000,000,000,000,000,000)
This equals roughly half the world’s biomass
How many viruses? (1x1031)
Whitman et al., 2003
Whitman et al., 1998; Anonymous 2011. Nature Rev Microbiol
… into context
…if we took a trowel-full (1
kg) of soil: ~50 billion
bacteria and arranged
them end-to-end
More than enough
bacteria to stretch from
the Queenstown to
Alexandra (~90 km)
But are microorganisms really
that important?
• Microbes are normally associated with
disease
But is this fair?
Almost every environment you can think of is
influenced or hosts a microbial community:
– N-fixation in plants
– Cow rumen
– Rocks (endoliths)
– Food
– Sewage
– Geothermal
– Human body
‘Everything is everywhere,
but the environment selects’
- Baas Becking, 1934
Extremophile
:[ɛksˈtrɛməfʌɪl]
(Ex.tre’mo.phile. L.n.
extremus, being on the
outside;
Gr.v. philos, loving.
Extremophile, extremityloving
Extremophily is an
anthropocentric concept
The Extremes of Temperature
?
120oC
100oC
Water boils
80oC
Hyperthermophiles
Hot springs
60oC
Scalding water
40oC
Body temperature
20oC
Too hot for most
NZer’s
0oC
Deep-sea
hydrothermal
vents
Water freezes
Thermophiles
Bath
Mesophiles
Surface water
Ice
Pyschrophiles
Extremophiles….
The Extremes of pH
0
0
4
Stomach acid
Lemon juice
2
Vinegar
4
6
6
Acid
2
pH
Pure water
8
Seawater
Volcanic crater
lakes
0
Acid springs
2
4
Neutral springs
8
10
Alkaline
Soap solution 10
Ammonia
12
12
Acidophiles
6
Neutrophiles
8
Alkaline springs
10
12
Alkaliphiles
An example of an extremophile
Pyrolobus fumarii
(Fire lobe of the chimney)
Grows best at 106 oC
Can grow as temps
as high as 113 oC
Stops growing if
temperature
goes below 90 oC!
Taq (Thermus aquaticus)
DNA polymerase
• Deinococcus –Thermus
• Isolated by Thomas Brock in 1969
– Yellowstone National Park
• Temp range 50-80°C (Topt 70°C)
• PCR – Taq Polymerase
the microbiology of geothermal hotsprings in New Zealand
J.Power
1000Springs.org.nz
facebook.com/1000SpringsProject
Aims and Objectives
To develop a comprehensive dataset of the
endemic microbial communities in New
Zealand’s geothermal ecosystems
objectives:
• Data collection (chemical, physical, and
biodiversity) of 1000 aqueous geothermal
ecosystems
• Publically available (Creative Commons 4.0):
Government agencies, regional councils,
tourism operators, schools, Māori groups,
scientific community
Consistent team and consistent application of
sampling/processing/analyses
Data collection and processing
• 1,019 features sampled
– 17 geothermal fields in the Taupō Volcanic
Zone
– 58 separate measurements per sample (not
including microbiology)
– 43K geochemical datapoints
– 15K metadata and geophysical datapoints
– 972 features sequenced for microbial
diversity
• 38.1M reads total;
• 32.2K different OTUs identified;
• 638 OTUs (av) per sample
White Island
19
Rotorua
378
Taheke
5
Tikitere
80
Waikite
44
Atiamuri
2
Te Kopia
21
Waimangu
107
Waiotapu
78
Whangairorohea
1
Orakei Korako
28
Ngatamariki
15
Wairakei-Tauhara
73
Tokaanu
40
Reporoa
29
Ohaaki
3
Rotokawa
74
additional outputs
• Fully interactive database
– search functions for multiple attributes including chemistry, physical
conditions, geothermal field/location, microbial taxonomy
• Publically-accessible web portal
(1000Springs.org.nz)
– includes common search functions, outreach, novel visualisations
of data, H&S and publications
• Software developed for field tablet
– software developed to capture metadata in-field to avoid
transcriptional errors and duplication
• Mobile Phone App in development
1,019 samples
13.9 – 100.6 oC
pH <0 – 9.7
38.1 M reads
32,200 OTUs
Average 638 OTUs
2L water
filtered
16S rRNA
gene amplicon
sequencing
- Ion Torrent
- EMP
primers
Mathew Button Duncan White
Alpha Diversity - Species Richness
Regional Community Composition
Beta Diversity - Bray Curtis
Stress = 0.22
Temp: ρ = 0.206
pH: ρ = 0.541
Waikite – Community Composition
Geochemistry
Stress = 0.22
Physicochemical knowledge put to use…
• Cultivation of thermophilic methanotrophs
Methylocystis
Methylococccus
Methylocaldum
Methylohalobius
Methylothermus
MDS2
0.50
0.00
0.25
temp
25
50
75
MDS2
100
pH
7.5
0.00
5.0
2.5
0.0
−0.25
−0.25
−0.25
0.00
MDS1
0.25
0.50
1000 Springs Project to
date
• Diversity peaks at low temperature,
circumneutral pH across the region
• pH is the main driver at regional
scale
• Where pH is static, temperature
takes over
• Hotsprings are dynamic
• temporal investigation needed
to understand true diversity
• Rare/novel strains present • But are they microbial kiwis?
Future plans
• Conservation and remediation
• Do we have endemic
microorganisms in NZ?
• Mātauranga
• Biogeography
• Targeted Bioprospecting
• Incorporate GIS and private user
login into the database
• Model for Biology National Science
Challenge database?
• Community function
• Metagenomics
• Can we predict community
composition from chemistry?
Acknowledgements
University of Waikato
Craig Cary
Ian McDonald
Georgia Wakerley
Charles Lee
Annika Hinze
Mathew Button
GNS Science
Jean Power
David Evans
Duncan White
Fieldwork Helpers
Karen Houghton
Carlo Carere
Funding
Kevin Lee
MBIE - NZ Government)
Hanna Peach
GNS Science core-funding Geothermal Resources of NZ
(GNS)