Smith et al – King crabs in Palmer Deep – Electronic Supplementary Material – Methods
Fig. S1-S2 and Tables S1-S2, Acknowledgments
METHODS
DNA Sequencing and Identification
Genomic DNA was extracted from muscle tissue using the Wizard SV Genomic DNA
Purification System (Promega) following manufacturer’s protocols. Polymerase chain reaction
(PCR) was performed in a 20 μl reaction volume containing 2.5 μl template DNA, 1X PCR
buffer (20 mM Tris, pH 8.8; 50 mM KCl; 0.01% Triton X-100, 0.02 mg/ml BSA), 2.5 mM
MgCl2, 0.125 mM dNTPs, 0.375 μM of each direction of the primer pair LCO1490/HCO2198
[31], and 0.2 μl (1 unit) of Taq polymerase (Bioline).
PCR was run on an Eppendorf
thermocycler as follows: 94 °C/3 min, 35 x (94 °C/15 s, 48 °C/15 s, 72 °C/30 s), 72 °C/5 min.
PCR product was purified using an ExoAP procedure and sequenced on a 3730xl DNA Analyzer
using Big Dye terminator chemistry (Applied Biosystems). A consensus sequence was produced
in Sequencher version 4.7 (Gene Codes) and compared against GenBank and the Barcode of Life
databases.
[31]
Folmer, O. Black, M. Hoeh, W. Lutz, R. & Vrijenhoek, R. 1994 DNA primers for
amplification of mitichondrial cytpchrome c oxidase subunit I from diverse metazoan
invertebrates. Mol. Mar. Biol. Biotechnol. 3, 294-299.
Figure S1. Phylogenetic relationships among species in the genus Neolithodes inferred from
577 nucleotides in the mitochondrial gene for COI using the neighbor-joining method. The tree
was made in MEGA version 5.05 from a bootstrap consensus of 10,000 replicates. Numbers
next to internal nodes indicate the percentage of replicate trees in which associated taxa clustered
together. Support values among individuals of the same species are not shown. Outgroups are
Lithodes confundens, Paralomis birsteini, and Pagurus bernhardus.
numbers are indicated in the tree.
GenBank accession
Figure S2. (A, B) Composite vertical across-shelf section, from the open slope to Palmer Deep,
of in situ temperature and density for summer (Dec-Feb) over the period 1982-2005. Locations
(dashed box) and approximate dates of profiles are shown in panel A. Dashed line in panel B
indicates the depth of the 1 °C isotherm. (C, D) Composite vertical across-shelf section, from
the open slope to Palmer Deep, of in situ temperature and density for winter (June-Aug) over the
period 1982-2005. Details as in panels A and B.
Table S1. Position, depth, date and time for important events observed along the survey during
ROV Dive # 10 on cruise NBP10-01 on the east side of Palmer Deep.
Event
Latitude
Longitude
Depth
(m)
1418.9
Date/ Time
(CLST)
02/24/2010
20:38
ROV arrival at seafloor (start of
transect)
64° 56’
56.0112S
64° 17’
44.6928W
Female Neolithodes yaldwyni
sampled
64° 56'
50.622S
64° 16'
16.251W
1154.8
02/24/2010
23:21
ROV position at 980m: last lithodid
crab observed on the transect
64° 56'
46.6296S
64° 15'
50.4606W
980.5
02/24/2010
23:59
ROV position at 850m depth contour:
no lithodid crab traces visible above
this depth
ROV departure from seafloor (end of
transect)
64° 15'
50.4606W
64° 15'
28.3644W
850.1
02/25/2010
00:24
64° 56’
38.5938S
64° 14’
29.9868W
601.1
02/25/2010
01:44
Table S2. Complete list of photographic and trawl surveys along the Antarctic Peninsula shelf at
depths of 430-725 m conducted during two major research projects: FOODBANCS2 on cruises
LMG02-08, NBP08-08 and LMG09-02 (C.R.S. Smith, L.J. Grange & C. Clark unpublished data
collected between 2008-2010) and LARISSA on cruise NBP10-01 (L.J. Grange & C.R.S. Smith
unpublished data collected in 2010). Figures in bold represent total survey area by location. No
lithodid crabs were detected during any of these surveys. Station locations are indicated in
Figure 1. For comparison, 9350 m2 were surveyed in Palmer Deep during ROV Dive # 10 for
this study.
Dates
Cruise
07/15/2008- NBP08-08
03/17/2009 LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
07/15/2008- NBP08-08
03/17/2009 LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
07/15/2008- NBP08-08
08/01/2008 LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
01/16/2010
Location
Station
AA
Station B
NBP10-01
~ Area
Depth
Latitude Longitude Operation
N surveyed
(m)
(m2)
Blake
578
63.05 S
61.59 W
3
11,300
Trawl
63.05 S
61.59 W
Otter
Trawl
578
5
37,500
63.05 S
61.59 W
YOYO
camera
578
9
2700
Stn AA Total
51,500
64.80 S
65.35 W
Blake
Trawl
578
2
7500
64.80 S
65.35 W
Otter
Trawl
578
6
45,000
64.80 S
65.35 W
SCUD
578
1
2500
64.80 S
65.35 W
YOYO
camera
578
9
2700
64.80 S
65.38 W
YOYO
camera
618
1
300
Stn B Total
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2008
LMG09-02
02/14/2008- LMG0803/17/2009
02,
Station E
58,000
65.98 S
67.28 W
Blake
Trawl
597
6
22,500
65.98 S
67.28 W
Otter
Trawl
597
1
7500
65.98 S
67.28 W
YOYO
camera
597
7
2100
Dates
Cruise
Location
~ Area
Depth
Latitude Longitude Operation
N surveyed
(m)
(m2)
NBP08-08
LMG09-02
Stn E Total
LMG0802/14/200802,
03/17/2009 NBP08-08
LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
66.98 S
69.71 W
Blake
Trawl
590
5
18,800
66.98 S
68.71 W
YOYO
camera
590
7
2100
Station F
Stn F Total
LMG0802/14/200802,
03/17/2009 NBP08-08
LMG09-02
LMG08-02
02/14/2008NBP08-08
03/17/2009
LMG09-02
NBP10-01
71.01 W
Blake
Trawl
582
6
22,500
68.13 S
71.01 W
YOYO
camera
582
6
1800
Station G
Admiralty
Sound
02/21/2010
01/21/2010
NBP10-01
Andvord
Bay
01/18/201002/20/2010
01/24/201001/25/2010
01/24/201001/27/2010
01/14/201001/18/2010
NBP10-01
NBP10-01
Barilari
Bay
Flandres
Bay
02/19/2010
02/20/2010
02/15/201002/18/2010
NBP10-01
Hughes
Bay
02/16/2010
02/08/2010
NBP10-01
20,900
68.13 S
Stn G Total
02/07/2010
32,100
Lockyer
64.42 S
57.66 W
64.82 S
62.64 W
64.77 S
62.82 W
64.82 S
62.64 W
65.94 S
64.64 W
65.78 S
64.86 W
65.00 S
63.32 W
64.24 S
61.36 W
64.18 S
61.22 W
64.24 S
61.21 W
64.17 S
61.22 W
64.47 S
57.80 W
ROV
486
1
survey
Blake
477
2
Trawl
ROV
430
1
survey
YOYO
509
8
camera
Andvord Bay Total
ROV
617
2
Survey
Blake
617
3
Trawl
Barilari Bay Total
YOYO
694
6
camera
Otter
526
1
Trawl
ROV
605
1
survey
YOYO
545
7
camera
YOYO
600
1
camera
Hughes Bay Total
Blake
469
1
24,300
4600
7500
7200
2400
17,100
5800
900
6700
1800
7500
3000
2100
300
12,900
3800
Dates
Cruise
Location
Station
02/07/2010
02/06/201002/08/2010
~ Area
Depth
Latitude Longitude Operation
N surveyed
(m)
(m2)
Trawl
ROV
64.47 S
57.83 W
725
2
3600
survey
YOYO
64.47 S
57.83 W
495
3
900
camera
Lockyer Station Total
8300
Grand Total 238,200