Supplementary Material Seasonal variation in abundance and diversity of bacterial methanotrophs in five temperate lakes Md Sainur Samad, Stefan Bertilsson* * Correspondence: Stefan Bertilsson,
[email protected] 1
Supplementary Figures and Tables
1.1
Supplementary Figures
Supplementary Figure S1 Location of the five Swedish lakes: Ekoln (59°46'N, 17°37'E), Erken (59°50'N, 18°35'E), Långsjön (60°01'N, 17°34'E), Siggeforasjön (59°58'N, 17°09'E) and Ramsen (59°49'N, 17°54'E) in Uppland, Sweden.
Supplementary Material
A
B
C
EkS
EkS
EkM EkB ErS ErM ErB LåS
EkM
EkB
EkS EkM EkB
ErS
ErM
Lå M LåB SiS SiM SiB RS
ErB
ErS ErM ErB
LåS
LåS
LåM
LåM LåB
LåB
L
L
RM
RB
C
L
SiS
SiM
SiS
SiB
SiM
RS
SiB
RM
RS
RM
RB
RB
L
L
Supplementary Figure S2 Agarose gel of PCR products (pmoA gene amplification without florescence primers [A189F, mb661R]) from all winter (A) samples. (B & C) PCR products (pmoA gene amplification with florescence primers [A189-HEX, mb661R]) from winter (B) and summer (C) samples. PCR amplicons were loaded on agarose after PCR purification kit (B & C). Abbreviations of Ek, Er, Lå, Si, R, S, M, B, C and L represent Ekoln, Erken, Långsjön, Siggeforasjön, Ramsen, surface, middle, bottom, negative control, and DNA ladder (TrackIt 100bp DNA Ladder) respectively.
2
Model quality by number of components 1
Index
0.8 0.6 0.4 0.2 0 Comp1
Q² cum
Comp2 Comp3 Components R²Y cum
Comp4
R²X cum
Supplementary Figure S3 PLS model quality by number of components (Comp 1 to 4).
3
Supplementary Material
VIPs (2 Comp / 95% conf. interval) 2.5
2
VIP
1.5
1
0.5
-0.5
-1
Temp. Season-S Season-W Phosphate Chl-a TDP Depth-B Oxygen DOC CH4 Color Sulfate Depth-S pH Depth-M
0
Variable
Supplementary Figure S4 The VIPs (Variable Importance for the Projection) for each explanatory variables of second component.
4
120
100
Winter
Summer
No. of TRFs
80
60
40
20
R (B)
R (M)
R (S)
Si (B)
Si (M)
Si (S)
Lå (B)
Lå (M)
Lå (S)
Er (B)
Er (M)
Er (S)
Ek (B)
Ek (M)
Ek (S)
0
Supplementary Figure S5 Total number of Terminal Restriction fragments (TRFs) of Methanotrophs in five Swedish lakes. Ek = Ekoln , Er = Erken, Lå = Långsjön , Si = Siggeforasjön, R = Ramsen, S = Surface, M = Middle and B = Bottom.
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Supplementary Material
1.2
Supplementary Table
Supplementary Table S1 Primers used in this study
Forward/
Target group
PCR
Sequence (5’ to 3’)a
Reference
GGNGACTGGGACTTCTGG/
Costello and
CCGGMGCAACGTCYTTACC
Lidstrom, 1999
length reverse primer (bP)
pmoA
A189F/
All methanotrophs
508
mb661R
Sequencing
M13F (-20)/
primers
GTAAAACGACGGCCAG/
(pmoA) of E. coli M13R
a
Inserted gene
CAGGAAACAGCTATGAC
N, bases A, C, T, or G; M, bases A or C; S, bases G or C; Y, bases C or T
2
Reference
Costello, A. M., and Lidstrom, M. E. (1999). Molecular characterization of functional and phylogenetic genes from natural populations of methanotrophs in lake sediments. Appl. Environ. Microbiol. 65, 5066–5074.
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