1
Erratum
2
3
Meares K, Dawson DA, Horsburgh GH, Perrin MR, Burke T, Taylor TD (2008)
4
Characterisation of 14 blue crane Grus paradisea (Gruidae, AVES) microsatellite
5
loci for use in detecting illegal trade. Conservation Genetics, in press.
6
7
This manuscript was published in Conservation Genetics in 2008 (online early) since publication
8
we have observed errors in the second part of Table 1 and Table 2.
9
10
The first part of Table 1 involving the characterization of the loci in blue crane did not contain
11
errors and remains unchanged.
12
13
The second part of Table 1 involving the cross-utility of the markers in other species, “Table 1
14
continued” which is highlighted in yellow, contains errors. In the published version of this part of
15
the table the samples sets of both species were contaminated by individuals from other species. The
16
grey-crowned cranes (Balearica regulorum) genotyped mistakenly included 3 blue crane
17
individuals and the wattled cranes (Grus carunculatus) genotyped included one blue crane and one
18
grey-crowned crane. The true number of alleles and heterozygosity observed for all of the 14 blue
19
crane loci when genotyped in grey-crowned and wattled crane is therefore much lower in these two
20
species than previously published. The species origins of the samples which were originally mis-
21
identified were confirmed by comparison of allele sizes between species using several loci which
22
displayed species-specific alleles (see Meares et al. 2008, submitted to Molecular Ecology
23
Resources).
24
25
After correction, eleven of the blue crane loci were polymorphic in grey-crowned crane (not 14) and
26
11 were polymorphic in wattled crane (not 14). Due to the reduced number of grey-crowned crane
27
and wattled crane individuals genotyped, tests for Hardy Weinberg equilibrium and estimated null
28
allele frequencies were not performed.
29
30
These errors have been rectified in the Table below.
31
32
Table 2 contains two minor errors. The numbers of grey-crowned cranes tested was 7 (not 10) and
33
the number of wattled cranes tested was 8 (not 10). These corrections have been highlighted in
34
yellow in the Table below.
35
36
We sincerely apologise for these errors.
1
37
38
39
Table 1 Characterisation of 14 blue crane (Grus paradisea) microsatellite loci in the blue crane, grey-crowned crane (Balearica regulorum) and wattled crane
(Grus carunculatus).
Blue crane (Grus paradisea)
Locus‡
EMBL
accession
number and
clone name
Primer sequence (5’ – 3’) with pigtail sequence underlined§
Gpa01
AM085152,
BC17E04
F: HEX - TTTGGTTGCCGTCAGAATTG
R: GTTTCTT-CTGACCTGGGTGTGTTCTGC
61.97
61.76
Gpa11†
AM282892,
BC43B09
F: HEX - CCCTCCTGGAATACATGACAAA
R: AAGTCATTCTCAACAAGAAAGGAAAA
Gpa12
AM282893,
BC43C11&
AM282894
AM282896,
BC43F06
Gpa24
MgCl2
(mM)
Repeat motif in
blue crane
n
A
Exp.
allele
size
(bp) ¥
Genotype of
library
source
individual
(J17718)
291, 297
Observed allele sizes (bp)
‡
HO
HE
63
1.5
(CA)8
19
4
297
(290)
291, 293, 297, 299
0.58
61.09
60.82
60
3.0
(TTTC)34
20
21
300
216, 236
186–346
F: FAM - GATCAATGCGAAGGATAGGGAGGT
R: TCATCAATCTATTATTTGCCTCAGC
61.70
60.78
60
3.0
(GATA)11
20
7
232,
236
232, 236
F: FAM - TTTCGTACTCTGGTCATTGGATTT
R: GTTTCTT-AATAGGACAGCAGTGCTAAGAAGAAA
61.07
61.07
57
2.0
(TTTC)12
20
12
283
(276)
AM282908,
BC45A12
F: HEX -GAGGGAATCTAGCACGCTCCAA
R: GTTTCTT-TCAAAGCATCGTGTCCATGAAGT
64.96
64.06
57
2.0
(GTTA)6
20
5
Gpa32
AM282918,
BC45H08
F: HEX -CCCAGCACACCGTGCATAAG
R: GTTTCTT-GCAGTCGGTCACATCCTTGG
64.85
64.02
57
2.0
(GT)11
20
Gpa33
AM282919,
BC45H10
F: FAM - GGCTTAGAAATGGGATACAGTTG
R: CCGTCCAAGCAAGAAGAAA
60.63
60.05
60
2.0
(TTTC)14
(TCTCTTTC)16
Gpa34
AM282920,
BC47A04
F: HEX -GCACACAGTGAGGAGACCAGTGA
R: CCTTGATGTGGGAAGACAACTGC
65.10
65.22
60
3.0
Gpa35
AM282921,
BC47A05
F: FAM - TCATCAGCTTCCAACAGGTCTCC
R: GTTTCTT-TCAGGCACAATGTATAAGTGTTTGTGG
65.01
65.09
57
Gpa36
AM282922,
BC47A12
F: HEX -TTCATAGACATATGCTTACCTGTTCT
R: ATCCATCCATCTATCTATCTATCTATCTATC
57.65
57.57
Gpa37
AM282923,
BC47D01
F: HEX -TGAACGTGTCTGATTTAAGGAA
R: AAAGTAGTCACTAGCCTGGGTTT
Gpa38
AM282924,
BC47H05
Gpa39
Gpa41
Gpa14
PRIMER3 Ta
Tm (oC)
(oC)
Est. null
allele
freq.
0.54
Deviation
from
HWE
(P-value
if >0.10)
NS
0.75
0.96
** (0.01)
0.11
212, 216, 220, 224, 228,
232, 236
0.75
0.83
NS
0.04
286, 294
278–324
0.85
0.90
NS
0.02
250
(243)
243, 251
239, 243, 247, 251, 255
0.50
0.62
NS (0.06) 0.12
7
190
(183)
183, 189
177, 183, 185, 187, 189,
191, 193
0.75
0.85
NS (0.07) 0.05
19
24
316
243, 243
205–417
0.90
0.97
NS
0.03
(TTTCTC)22
17
18
265
NA
249–413
0.88
0.95
NS
0.02
2.0
(GT)13 (TA)4
18
7
176
(169)
176, 178
172, 174, 176, 178, 180,
182, 184
0.89
0.81
NS
-0.07
60
2.0
(GATA)10
(GATG)2
19
5
228
228, 228
220, 224, 228, 232, 236
0.53
0.67
NS
0.10
57.43
57.64
60
2.0
19
27
231
194, 194
174–390
0.95
0.98
NS
0.00
F: HEX -GGGCAGAAGCAAGTCTTTCA
R: GAAGATGTTTGCTGGTTGCAC
60.52
60.69
60
3.0
(TC)2 CTC
(TTTC)43
(TTC)2
(CTAT)13
20
6
193
188, 192
180, 184, 188, 192, 196,
200
0.90
0.81
NS
-0.06
AM282925,
BC47H08
F: HEX -TGCACAGGTTTGGCCAAGAAG
R: GTTTCTT-TTCCAAAGTGAAATTAAAGGTGTGTGG
65.66
65.13
57
2.0
(GA)2
(GATA)13
20
7
125
(118)
122, 126
114, 118, 122, 126, 130,
134, 138
0.85
0.81
NS
-0.04
AM282932,
BC49F10
F: FAM - AATTCAGTCTAATGTAAATGTCCAAG
R: CCATCATTAATGGAAAGAAAGAG
57.12
56.99
54
2.0
(TTTCTTCC)6
(TC)5
(TTTCTCTC)3
(TTTCTCTC)3
(TTTC)2
20
4
242
245, 245
237, 245, 253, 261
0.60
0.60
NS
0.00
-0.04
40
2
Table 1 continued
Grey-crowned crane (Balearica regulorum)
Wattled crane (Grus carunculatus)
Locus‡
TD temp.
(oC)
n
A
Allele sizes (bp) ‡
HO
HE
Deviation
from HWE
n
A
Allele sizes (bp) ‡
HO
HE
Deviation
from HWE
-
Est.
null
allele
freq.
-
8
1
291
0.00
0.00
-
Est.
null
allele
freq.
-
Gpa01
66-60
7
1
293
0.00
0.00
Gpa11
63-57
7
7
234, 248, 258, 262, 270, 274, 310
1.00
0.90
NA
NA
8
9
224, 230, 245, 248, 254, 258, 266,
282, 294
0.88
0.93
NA
NA
Gpa12
63-57
7
3
196, 200, 204
0.43
0.63
NA
NA
8
3
192, 196, 200
0.75
0.63
NA
NA
Gpa14
60-54
6
6
246, 250, 254, 262, 266, 270
0.67
0.88
NA
NA
8
5
270, 294, 298, 308, 358
0.38
0.81
NA
NA
Gpa24
60-54
1
2
243, 247
1.00
1.00
NA
NA
8
1
243
0.00
0.00
-
-
Gpa32
60-54
2
1
177
0.00
0.00
-
-
8
2
179, 181
0.50
0.50
NA
NA
Gpa33
63-57
7
3
206, 214, 218
0.29
0.56
NA
NA
5
8
218, 254, 314, 318, 326, 334, 336,
344
1.00
0.96
NA
NA
Gpa34
63-57
7
7
132, 138, 144, 152, 164, 176, 220
0.86
0.85
NA
NA
7
7
280, 286, 292, 298, 304, 320, 322
0.86
0.87
NA
NA
Gpa35
60-54
7
2
159, 161
0.14
0.36
NA
NA
8
1
169
0.00
0.00
-
-
Gpa36
63-57
7
3
228, 232, 236
0.57
0.56
NA
NA
8
4
220, 224, 228, 232
1.00
0.70
NA
NS
Gpa37
63-57
2
2
197, 205
0.50
0.50
NA
NA
8
8
121, 123, 129, 131, 133, 137, 141,
147
0.88
0.88
NA
NA
Gpa38
63-57
2
1
189
0.00
0.00
-
-
7
4
181, 185, 193, 197
1.00
0.67
NA
NA
Gpa39
60-54
7
9
176, 198, 202, 206, 216, 222, 224,
234, 250
0.57
0.90
NA
NA
8
2
102, 106
0.13
0.13
NA
NA
Gpa41
57-51
7
7
187, 191, 207, 211, 215, 219, 227
1.00
0.88
NA
NA
8
4
233, 249, 257, 265
0.38
0.35
NA
NA
3
Tm, Melting temperature assigned by PRIMER v3 software (Rozen et al. 2000);
Ta, annealing temperature;
n, number of crane individuals that successfully amplified and were genotyped of the 20 blue crane, 10 wattled crane and 10 grey-crowned crane individuals tested, respectively;
A, number of alleles;
¥, expected allele size based on the sequenced clone including the 7-bp length of the pigtail when used. The expected allele size excluding the length of the pigtail is shown in parentheses;
HO, observed heterozygosity calculated using CERVUS v2.0 (Marshall et al. 1988);
HE, expected heterozygosity calculated using CERVUS v2.0 (Marshall et al. 1988);
Deviation from HWE, deviations from Hardy-Weinberg equilibrium (HWE) calculated using GENEPOP 3.4 (Raymond, 1995);
Est. null allele freq., estimated null allele frequency calculated using CERVUS v2.0 (Marshall et al. 1988);
†, locus has high predicted null allele frequency of equal or above 0.1;
§, for some loci a “pigtail” sequence GTTTCTT added, to reduce noise from variable adenylation during the PCR (Brownstein et al. 1996);
‡, the seven bases of the pigtail are included in the observed allele size;
NS, non-significant; NA, not available; *significant at the 5% level, **significant at the 1% level;
TD temp., maximum and minimum temperatures used in touchdown PCR. Touchdown PCR program used: 94°C for 3 min; 5 cycles of 94°C for 30 s, highest T a, decreasing with 1°C increments per
cycle, 72°C for 30 s; followed by 25 cycles of 94°C for 30 s, lowest T a for 30 s, 72°C for 30 s, completing the profile with an extension at 72 °C for 4 min;
‡Locus Gpa26 (clone BC45E05, AM282912) was monomorphic in 76 blue crane individuals but was not tested in wattled crane and grey-crowned crane. Primer sequences for this locus are F:
AACTAGGTTAACAGATGCATTAACAGG, R: GGAAGTGGCGGGTATAGGAA.
Sources of individuals genotyped:
Blue crane individuals originated from a wild population inhabiting the Karoo region, South Africa;
Wattled crane individuals originated from KwaZulu-Natal South Africa;
Grey-crowned crane individuals originated from KwaZulu-Natal, Eastern Cape and Mpumalanga provinces, South Africa (population structure currently uncertain).
4
Table 2 Cross-species utility of 14 blue crane Grus paradisea microsatellite loci.
Species
Binomial name
Order
n
Genetic
distance to
blue crane‡
Gpa01
Gpa11
Gpa12
Gpa14
Gpa24
Gpa32
Gpa33
Gpa34
Gpa35
Gpa36
Gpa37
Gpa38
Gpa39
Gpa41
Grey-crowned crane*
Balearica regulorum
Gruiformes
7
0.7
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Wattled crane*
Grus carunculatus
Gruiformes
8
0.7
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Houbara bustard†
Chlamydotis undulata
Gruiformes
1
16.9
+
+
–
–
–
–
–
–
–
–
–
–
–
–
Grey-headed albatross†
Diomedea chrysostma
Procellariiformes
1
20.1
–
+
–
–
–
–
–
–
+
–
+
+
+
–
Seychelles warbler†
Acrocephalus sechellensis
Passeriformes
1
21.6
–
–
–
–
–
–
–
–
–
–
–
–
–
–
Cape parrot†
Poicephalus robustus
Psittaciformes
2
23.1
+
+
–
–
+
+
–
–
–
–
–
–
–
–
Red jungle-fowl†
Gallus gallus
Galliformes
1
28.0
–
–
–
–
–
–
–
–
–
–
–
–
–
–
Saltwater crocodile†
Crocodylus porosus
Crocodilia
1
NA
–
–
–
–
–
–
–
–
–
–
–
–
–
–
Human†
Homo sapiens
Primates
2
NA
–
–
–
–
–
–
–
–
–
–
–
–
–
–
n, number of individuals tested for amplification;
+, amplification observed on 2% agarose gel;
–, no amplification observed on 2% agarose gel, or non-specific amplification or amplification of a product over 200 bp larger or smaller than the size observed in blue crane;
*amplified using touchdown PCR (temperature range provided in Table 1) and genotyped using an ABI 3730 DNA Analyzer;
†, amplified using same PCR conditions as blue crane and visualised on a 2% agarose gel;
‡, delta T50H DNA–DNA hybridisation distances based on Sibley et al. (1990);
NA, not available.
5