Craigiebuckler, Aberdeen, AB15 8QH, UK

RECIPE Charquemont Progress meeting

23 - 26/10/2003

UK progress

Rebekka Artz

Stephen Chapman

Colin Campbell

I. WP01 UK Field site

• Middlemuir Moss, between Strichen and New Pitsligo, NE Scotland

Bare milled peat (0-5 yrs)

1st succession peat (> 5yrs)

Regenerating peat (> 50 yrs)

> 50 yrs 5 -10 yrs Bare

Table 1. Approximate species distribution at RECIPE WP01 field sites

Experimental Site / Keystone species

Plant distribution (%)

Collar 1 Collar 2 Collar 3

Bare milled surface

bare bare bare

1st Succession Peat / Eriophorum

EA (80) SF (15) EV (<5) CI (<1)

EA (80)SF (15)EV (5)

EA (70)SF (20)EV (5)CS (3)CI (2)

1st Succession Peat / Sphagnum

SF (100) SF (95) EA (5)

SF (95) EA (5)

Regenerating Peat / Polytrichum

PC (80)SC (10)HS (3)PS (3)CS (4)

PC (75)SM (5)SC (10)SCU (5)CS (5)

PC (95)SM (<1)SC (<4)O

Regenerating Peat / Sphagnum

SC (80)SCU (15)SM (5)

SC (60)SCU (15)SM (5)HS (10)PS (5)CS (<5)O

SC (70)SCU (15)SM (5)PS (5)CS (<5)O

Key: EV – Eriophorum vaginatum,EA – Eriophorum angustifolium,SF – Sphagnum fallax,SM – Sphagnum magellanicum,SC – Sphagnum capillifolium,SCU – Sphagnum cuspidatum,CI – Campylopus introflexus,PC – Polytrichum commune, HS – Hypnum sp. (jutlandicum or cypressiforme),PS – Pleurozium schreberi, CS – Calluna sp., O – others (grasses etc.)

II. Field site preliminary data

Water table in plotsBare peat

-120

-100

-80

-60

-40

-20

0

0 10 20 30 40 50 60 70

time (d)

wat

er t

able

(cm

)

well 1 well 2 well 3 1st succession peat

-120

-100

-80

-60

-40

-20

0

0 10 20 30 40 50 60 70

time (d)

wa

ter

tab

le (

cm

)

well 1 well 2 well 3

Regenerating peat

-120

-100

-80

-60

-40

-20

0

0 10 20 30 40 50 60 70

time (d)

wat

er t

able

(cm

)

well 1 well 2 well 3

Chamber leak test - no. I

0

200000

400000

600000

800000

1000000

1200000

1400000

1600000

1800000

-3 0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48

time (min)

(CH

4 ar

ea)

Methane added

Field measurements of CO2 and CH4 fluxes

Bare peat – CO2

DARK 25/09

0

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0 5 10 15 20 25 30

time (min)

CO

2 (%

)

bare 1 bare 2 bare 3 DARK 16/10

0

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0 5 10 15 20 25 30

time (min)

CO

2 (%

)

bare 1 bare 2 bare 3

- 41 cm- 90 cm

Bare peat – CH4

DARK 25/09

0

1

2

3

4

5

0 10 20 30time (min)

CH

4 in

pp

m

collar no 1 collar no 2 collar no 3

- 90 cm

DARK 16/10

0

1

2

3

4

5

0 5 10 15 20 25 30

time (min)

CH

4 in

pp

m

bare 1 bare 2 bare 3

- 41 cm

1st succession peat – CO2

DARK 25/09

0

0.02

0.04

0.06

0.08

0.1

0.12

0.14

0.16

0.18

0 5 10 15 20 25 30time (min)

CO

2 (%

)

Sph 1 Sph 2 Sph 3 Erio 1 Erio 2 Erio 3

- 22 cm

DARK 16/10

0

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0 5 10 15 20 25 30

time (min)

CO

2 (%

)

Sph 1 Sph 2 Sph 3 Erio 1 Erio 2 Erio 3 LIGHT 16/10

0

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0 5 10 15 20 25 30time (min)

CO

2 (%

)

Sph 1 Sph 2 Sph 3 Erio 1 Erio 2 Erio 3

- 7 cm - 7 cm

LIGHT 25/09

0

0.05

0.1

0.15

0.2

0.25

0.3

0 5 10 15 20 25 30time (min)

CO

2 (%

)

Sph 1 Sph 2 Sph 3 Erio 1 Erio 2 Erio 3

- 22 cm

1st succession peat – CH4

DARK 25/09

0.01.02.03.04.05.06.07.08.09.0

10.0

0 5 10 15 20 25 30

time (min)

CH

4 in

pp

m

Sphagnum 1 Sphagnum 2 Sphagnum 3Eriophorum 1 Eriophorum 2 Eriophorum 3

LIGHT 25/09

0.01.02.03.04.05.06.07.08.09.0

10.0

0 5 10 15 20 25 30

time (min)

CH

4 i

n p

pm

Sphagnum 1 Sphagnum 2 Sphagnum 3

Eriophorum 1 Eriophorum 2 Eriophorum 3

- 22 cm - 22 cm

DARK 16/10

0.00.51.01.52.02.53.03.54.04.55.0

0 5 10 15 20 25 30time (min)

CH

4 i

n p

pm

Sphagnum 1 Sphagnum 2 Sphagnum 3

Eriophorum 1 Eriophorum 2 Eriophorum 3

- 7 cm

LIGHT 16/10

0.00.51.01.52.02.53.03.54.04.55.0

0 5 10 15 20 25 30time (min)

CH

4 in

pp

m

Sphagnum 1 Sphagnum 2 Sphagnum 3

Eriophorum 1 Eriophorum 2 Eriophorum 3

- 7 cm

Regenerating peat – CO2

DARK 30/09

0

0.005

0.01

0.015

0.02

0.025

0.03

0.035

0.04

0 5 10 15 20 25 30

time (min)

CO

2 (%

)

Sph 1 Sph 2 Sph 3 Poly1 Poly2 Poly3 LIGHT 30/09

-0.2

-0.15

-0.1

-0.05

0

0.05

0 5 10 15 20 25 30

time (min)

CO

2 (%

)

Sph 1 Sph 2 Sph 3 Poly1 Poly2 Poly3

- 38 cm - 38 cm

Regenerating peat – CH4

DARK 30/09

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

0 5 10 15 20 25 30time (min)

CH

4 in

pp

m

Sphagnum 1 Sphagnum 2 Sphagnum 3

Polytrichum 1 Polytrichum 2 Polytrichum 3

LIGHT 30/09

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

0 10 20 30

time (min)C

H4

in p

pm

Sphagnum 1 Sphagnum 2 Sphagnum 3

Polytrichum 1 Polytrichum 2 Polytrichum 3

- 38 cm - 38 cm

III. Fungal community structure analysis

by DGGE of ribosomal 18S and/or ITS sequences

Preliminary data

Test DNA extractions for shipping methods

0

10

20

30

40

50

60

Living sphagnumlayer

Low humificationpeat

Medianhumification peat

High humificationpeat

Milled surfacepeat

mg

ml-1

DN

A

NO TREATMENT LIQUID N2 FREEZE-DRIED

***

*p < 0.05

M + NT/S LN/S FD/S NT/P1 LN/P1 FD/P1 Sphagnum Low humification

+ M NT/P2 LN/P2 FD/P2 NT/E LN/E FD/E + M

Medium humification High humification

M + P2old P2new NT/M LN/M FD/M + + M

PCR FD/P2 Milled surface

M + NT LN FD Regenerating S E

Fungal DGGE analysis shows significant differences in band patterns according to peat depth

but: GELS CANNOT BE INTERCOMPARED (YET)

IV. Community level physiological profiling

by MicroResp

Preliminary data

• ‘Standard’ method utilises sieved soil of ca. 40% WHCproblematic with peat due to irreversible drying and fibrosity

• Protocol adapted to use slurry (peat blended with water 1:10)

• tested with glucose (150 and 75 mM)

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0 200 400 600 800 1000 1200 1400

time (min)

A 5

90

nm

20-30 bare 10-20 bare 5-10 bare 0-5 bare

20-30 >5 10-20 >5 5-10 >5 0-5 >5

20-30 >50 10-20 > 50 5-10 >50 control

Blank

only the surface sample at Regenerating peat site gives significantly higher SIR than a water blank (not sterile). No distinction possible between samples so far.

150 mM glucose, 2.5 mM bicarb

2.5 mM

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0 5 10 15 20 25time (h)

A 5

90

nm

2.0 mM0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0 5 10 15 20 25

time (h)

A 5

90

nm

1.5 mM0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0 5 10 15 20 25time (h)

A 5

90 n

m

1.0 mM0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0 5 10 15 20 25time (h)

A 5

90

nm

75 mM glucose, varying bicarb

0.5 mM

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0 5 10 15 20 25

time (h)

A 5

90 n

m

All 4 peat depths at Regenerating site, 0-5 cm horizon at 1st succession site, and all except the 0-5 cm horizon at the bare site show significantly higher CO2 evolution than the (water only) control. Different samples show different CO2 curves.

Lowering the buffering capacity of the detection system allows for sample distinction.

Further work:

incorporation of different carbon sources to allow for CLPP using 32 carbon sources per plate (n = 3) or 16 carbon sources (n = 6).

Anoxic system?

V. RECIPE WEBSITE

Preliminary version available at

http://www.macaulay.ac.uk/recipe

Will go public soon …