apidologie

Supplementary Material
APIDOLOGIE
A scientific note on an anomalous diploid individual of Euglossa melanotricha (Apidae,
Euglossini) with both female and male phenotypes
Karen M. Suzuki, Douglas C. Giangarelli, Dhiego G. Ferreira, Wilson Frantine-Silva, Solange
C. Augusto, Silvia H. Sofia*
*Corresponding author: S.H. Sofia
E-mail: [email protected]
Laboratório de Genética e Ecologia Animal, Departamento de Biologia Geral, CCB,
Universidade Estadual de Londrina, Rodovia Celso Garcia Cid, km 380, 86057-970,
Londrina, PR, Brazil
Table I-S. Primer annealing temperatures and PCR conditions used in the amplification of
the seven microsatellite loci for the orchid bee Euglossa melanotricha. Reagents
DNA template
GoTaq Master Mix (Promega)
glycerol
tailed primer
forward primer
reverse primer
ultrapure H2O
Amplification conditions
initial step
10 cycles
Concentration (volume)
5 ng
1x
1%
0.125 µM
0.0125
0.125 µM
(up to 5 µL)
Temperature/time
95°C/5min
[94°C/30s; primer annealing temp/1 min; 72°C/1min]
25 cycles
[89°C/30s; primer annealing temp/1 min; 72°C/1min]
final extension
primer annealing
(Temperature oC)
72°C/30 min
ann24, ann37 and
ann2 and Egc37 Egc26 and ann8
ann41
(53oC)
(54oC)
o
(56 C)
F and R: forward and reverse primers, respectively. Forward primers of each pair received a
small
additional
5’end
tag.
Tailed
primer
=
M13
sequence
5'TGTAAAACGACGGCCAGT-3' labelled with fluorophores Table II-S. Alleles identified at the seven microsatellites loci analyzed for hind leg samples (left/male and right/female) of the anomalous individual
(‘gynandromorph’) of Euglossa melanotricha, showing the phenotypes of both sexes, and for samples (legs) of the two control females (F1 and F2)
exhibiting normal phenotypes. F1 and F2 correspond to females from the same nest of the gynander, however from different generations. M1 and M2
(control) correspond to two normal males of E. melanotricha surveyed in different localities from gynander and the two control females (F1 and F2),
which were used for comparative purpose. Alleles in bold correspond to those alleles commons to bees (anomalous and both females F1 and F2) from
the same nest, but from different generations.
Alleles found for each locus
Alleles found for each sample
Size
Left hind leg
Right hind leg
Control
Control
Control
Control
(pb)
(male phenotype)
(female phenotype)
F1
F2
M1
M2
(186-200)a
192/198
192/198
174/198
174/198
174
174
(165-197)a
166/170
166/170
160/168
166/168
162
160
(174-177)a
158/161
158/161
158/161
158/161
158
158
(152-166)a
174/178
174/178
134/178
134/178
134
134
(164-176)a
154
154
154
154
154
154
(164-168)b
144/162
144/162
159/162
159/162
162
162
(154-166)b
156
156
140/156
156/159
149
149
Locus
ann02
a
ann24
a
ann28
a
ann37
a
ann41a
Egc 26b
Egc 37b
a
Paxton et al. (2009) estimated for E. annectans; bSouza et al. (2007) tested on E. melanotricha.