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Supplementary Information
Impact of a simulated oil spill on benthic phototrophs and nitrogen-fixing bacteria in
mudflat mesocosms.
Panagiota-Myrsini Chronopoulou1†, Anne Fahy1‡, Frédéric Coulon1,6, Sandrine Païssé2,
Marisol Goñi-Urriza2, Louis Peperzak3, Laura Acuña Alvarez1, Boyd A. McKew1, Tracy
Lawson1, Kenneth N. Timmis4,5, Robert Duran2, Graham J. C. Underwood1 and Terry J.
McGenity1*
1 School
of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK.
2 Equipe
Environnement et Microbiologie, UMR CNRS-IPREM 5254, IBEAS, Université de Pau
et des Pays de l’Adour, Pau, BP 1155 F-64013, France.
3
Department of Biological Oceanography, Royal Netherlands Institute for Sea Research
(NIOZ), PO Box 50, NL 1790, AB Den Burg, The Netherlands.
4 Helmholtz
5 Institute
6
Center for Infection Research, Braunschweig, Germany.
for Microbiology, Technical University, Braunschweig, Germany.
Department of Environmental Science and Technology, School of Applied Sciences,
Cranfield University, Cranfield, MK43 0AL, UK.
* For
correspondence: Dr. Terry J. McGenity, Department of Biological Sciences, University of
Essex, Wivenhoe Park, Colchester, CO4 3SQ, UK. Email: [email protected]
†
Present address: School of Biological and Chemical Sciences, Queen Mary University of
London, Mile End Road, London, E1 4NS, UK.
‡
Present address: Equipe Environnement et Microbiologie, UMR CNRS-IPREM 5254, IBEAS,
Université de Pau et des Pays de l’Adour, Pau, BP 1155 F-64013, France.
Running Title: Cyanobacterial bloom in oiled mudflat mesocosms
Keywords: wetland, mudflat, salt marsh, crude oil, biodegradation, nitrogen fixation, nifH,
diatoms, cyanobacteria, biofilm, hydrocarbon
Supplementary Table
Table S1. Primers used for amplification of nifH gene
Primer name Positions
Sequence (5´ - 3´)
Reference
nifH1
639 to 655
TGY GAY CCN AAR GCN GA
Zehr and McReynolds (1989)
nifH2
1000 to 984
ADN GCC ATC ATY TCN CC
Zehr and McReynolds (1989)
nifH4
546 to 562
TTY TAY GGN AAR GGN GG Zani et al. (2000)
nifH3
1018 to 1002 ATR TTR TTN GCN GCR TA
Zani et al. (2000)
Positions refer to Azotobacter vinelandii nucleotide positions. The primer set nifH1/nifH2 was
constructed for amplification of a 359 bp fragment and primers nifH4/nifH3 for amplification of a
460 bp fragment of the nifH gene.
Supplementary Figures
A
Plan view
Tanks containing
sediment cores
Reservoir tanks
with pump
Treatments
COK = Colne Oil Killed
COL = Colne Oil Live
CL = Colne Live
B
Side view (low tide)
pump
Side view (high tide)
pum p
Fig. S1. Schematic representation of the experimental tidal mesocosms as described in
detail by Coulon et al. (2012). (A) Plan view showing the nine containers (32.5 x 42 x 25.5
cm) holding 20 sediment cores (9.8 cm high, 6.5 cm diameter) that were positioned above
reservoir tanks. The three different treatments are indicated. (B) Side view showing the
upper container holding the sediment cores and the lower reservoir, with the pump
illustrating the flow of seawater.
Fig. S2. Solar irradiance experienced by the sediment during the course of the
experiment. Two tube solar energy sensors (Solm) were placed at either end of the tanks
used to hold the sediment cores (see Coulon et al. (2012) for details).
Fig. S3. Examples of fluorescence micrographs. Chlorophyll a fluorescence (F’; A, C, E, left
column) and photosystem II operating efficiency (as measured by Fq’/Fm’; B, D, F, right
column), where green = least efficient, blue = intermediate, red = most efficient. Figures A
and B are from an oiled day-7 sediment core, Fq’/Fm’ = 0.112. Figures C and D are from an
oiled day-14 sediment core, Fq’/Fm’ = 0.172. Figures E and F are from a non-oiled day-14
sediment core, Fq’/Fm’ = 0.222. Each field of view is 640  480 µm.