A systematic study on Endotribelos Grodhaus (Diptera: Chironomidae) from Brazil including DNA barcoding to link males and females Author Trivinho-Strixino, Susana Author Pepinelli, Mateus text Zootaxa 2015 3936 1 1 41 journal article 10.11646/zootaxa.3936.1.1 1745dfc0-1683-48d2-9389-1da2d885ddaf 1175-5326 288149 14342F0D-B747-467B-A1B3-5FD7264A459F Endotribelos bicolor sp. n. ( Figs 1–4 ) Type material . Holotype : Male with pupal and larval exuviae, Brazil , São Carlos, SP, Represa do Fazzari, 20.iv.2014 , S. Trivinho-Strixino, (E3-07, LEIA-UFSCar). Paratypes : 1 male with pupal exuvia, in two slides mounted in Euparal, Brazil , Luiz Antônio, SP, Lagoa do Óleo, 25.vii.2003 , (E3-08, E3-09, LEIA-UFSCar); 1 pupa with pharate female, Luiz Antônio, SP, Represa do Beija-Flor, xi.2011 (E3-10, LEIA-UFSCar); 1 female with pupal and larval exuviae, same locality; 04.x.2001 (E3-11, LEIA-UFSCar). Etymology. From Latin, bicolor , referring to the brown and yellow coloration of adult male. TABLE 1. List of specimens of Endotribelos with mtDNA COI sequences (DNA Barcoding) known.
Taxon Locality Endotribelos fulvidus sp. n. female BRAZIL, São Carlos, Córrego do 29, 22 may 2012 Collection reference BNB2-00726 Coordinates -21,9367; -47,9042 Altitude GenBankacession number 685 m.a.s.l. KP237049
Endotribelos fulvidus sp. n. male BRAZIL, São Carlos, Córrego do 29, 23 may 2012 BNB2-00727 -21,9367; -47,9042 685 m.a.s.l. KP237041
Endotribelos euterpe male BRAZIL, São Carlos, Córrego do 29, 26 may 2012 BNB2-00729 -21,9367; -47,9042 685 m.a.s.l. KP237048
Endotribelos sublettei sp. n. female BRAZIL, São Carlos, Córrego Espraiado, 3 may 2012 BNB2-00732 -21,8972; -47,8736 830 m.a.s.l. KP237039
Endotribelos calophylli female BRAZIL, São Carlos, Córrego Espraiado, 20 april 2012 BNB2-00733 -21,8972; -47,8736 685 m.a.s.l. KP237038
Endotribelos fulvidus sp. n. male BRAZIL, São Carlos, Córrego do 29, 1 may 2012 BNB2-00734 -21,9367; -47,9042 685 m.a.s.l. KP237037
Endotribelos calophylli male BRAZIL, São Carlos, Córrego do 29, 30 april 2012 BNB2-00735 -21,9367; -47,9042 685 m.a.s.l. KP237036
Endotribelos sublettei sp. n. male BRAZIL, São Carlos, Córrego do 29, 5 may 2012 BNB2-00736 -21,9367; -47,9042 685 m.a.s.l. KP237050
Endotribelos fulvidus sp. n. male BRAZIL, São Carlos, Córrego do 29, 1 may 2012 BNB2-00737 -21,9367; -47,9042 685 m.a.s.l. KP237045
Endotribelos fulvidus sp. n. male BRAZIL, São Carlos, Córrego do 29, 1 may 2012 BNB2-00738 -21,9367; -47,9042 685 m.a.s.l. KP237044
Endotribelos fulvidus sp. n. female BRAZIL, São Carlos, Córrego do 29, 1 may 2012 BNB2-00739 -21,9367; -47,9042 685 m.a.s.l. KP237043
Endotribelos semibruneus sp. n. BRAZIL , São Carlos, Córrego do Espraiado, 3 may 2012 BNB2-00740 -21,8972; -47,8736 830 m . a.s.l. KP237047 female Endotribelos fulvidus sp. n. female BRAZIL , São Carlos, Córrego do 29, 1 may 2012 BNB2-00741 -21,9367; -47,9042 685 m . a.s.l. KP237046 Endotribelos fulvidus sp. n. female BRAZIL , São Carlos, Córrego do 29, 1 may 2012 BNB2-00744 -21,9367; -47,9042 685 m . a.s.l. KP237040 TABLE 2. Estimates of evolutionary divergence between sequences of Endotribelos species. The number of base substitutions per site from between sequences are shown. Analyses were conducted the Kimura 2-parameter model and conducted in MEGA6 software.
Endotribelos semibruneus sp. n. male Endotribelos euterpe male BRAZIL, São Carlos, Córrego do Espraiado, 10 sept 2010 BRAZIL, São Carlos, Córrego do Espraiado, 10 sept 2010 MPSW-000116 MPSW-000118 -21,8972; -47,8736 -21,8972; -47,8736 830 m.a.s.l. 830 m.a.s.l. KP237035 KP237033
Endotribelos euterpe female BRAZIL, São Carlos, Córrego do Espraiado, 10 sept 2010 MPSW-000120 -21,8972; -47,8736 830 m.a.s.l. KP237034
Endotribelos cf. hesperium ? GU565739
Endotribelos cf. hesperium ? GU565821
Endotribelos sp. ? GU565751
Endotribelos sp. ? GU565830
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22
E. euterpe E. euterpe E. semibruneus E . cf hesperium E . sp . E . cf hesperium E . sp . E . fulvidus E . fulvidus E . fulvidus E . semibruneus E . fulvidus E . fulvidus E . fulvidus E . sublettei E . calophylli E . fulvidus E . calophylli E . sublettei E . euterpe E . fulvidus E . sublettei
female male male GU565739 GU565751 GU565821 GU565830 male female female female female male male male male male female female male male female
0.012
0.174 0.176
0.162 0.157 0.167
0.177 0.180 0.153 0.188
0.169 0.171 0.188 0.193 0.158
0.130 0.130 0.151 0.144 0.152 0.134
0.142 0.147 0.168 0.135 0.151 0.180 0.137
0.140 0.146 0.170 0.135 0.151 0.183 0.137 0.002
0.136 0.142 0.172 0.140 0.145 0.178 0.134 0.008 0.006
0.153 0.155 0.022 0.167 0.153 0.147 0.151 0.148 0.150 0.151
0.142 0.147 0.168 0.135 0.151 0.180 0.137 0.000 0.002 0.008 0.148
0.142 0.147 0.168 0.135 0.151 0.180 0.137 0.000 0.002 0.008 0.148 0.000
0.142 0.147 0.168 0.135 0.151 0.180 0.137 0.000 0.002 0.008 0.148 0.000 0.000
0.174 0.174 0.185 0.179 0.174 0.169 0.131 0.147 0.149 0.149 0.164 0.147 0.147 0.147
0.131 0.129 0.163 0.153 0.172 0.146 0.134 0.155 0.157 0.157 0.142 0.155 0.155 0.155 0.172
0.136 0.142 0.172 0.140 0.145 0.178 0.134 0.008 0.006 0.000 0.151 0.008 0.008 0.008 0.149 0.157
0.133 0.131 0.163 0.153 0.172 0.145 0.137 0.155 0.157 0.157 0.142 0.155 0.155 0.155 0.172 0.002 0.157
0.172 0.170 0.181 0.182 0.172 0.164 0.127 0.147 0.149 0.149 0.159 0.147 0.147 0.147 0.006 0.168 0.149 0.168
0.014 0.014 0.170 0.157 0.172 0.166 0.127 0.140 0.138 0.135 0.149 0.140 0.140 0.140 0.167 0.129 0.135 0.131 0.163
0.142 0.147 0.168 0.135 0.151 0.180 0.137 0.000 0.002 0.008 0.148 0.000 0.000 0.000 0.147 0.155 0.008 0.155 0.147 0.140
0.140 0.146 0.226 0.135 0.151 0.207 0.137 0.002 0.000 0.006 0.150 0.002 0.002 0.002 0.149 0.157 0.006 0.157 0.149 0.138 0.002
FIGURE 1 . Endotribelos bicolor sp. n. Male. a . Thorax, dorsal view. b . Thorax, lateral view. c . Abdomen dorsal view. d . Front, mid and hind tibial spurs. e. Hypopygium, left dorsal view, right dorsal view tergite IX removed and with details of left upper volsella. f . Superior volsella. FIGURE 2. Endotribelos bicolor sp. n. Female. Genitalia ventral. Diagnostic characters. Endotribelos bicolor sp. n. differs from all other species of the genus by the combination of the following characters. Adult male : thorax yellowish with brown mesonotal stripes and short brown triangular spot before the scutellum; scutellum pale; postnotum brown; abdomen with uniform brown coloration; anal point with apex larger than base and shorter than inferior volsella. Pupa : abdominal posterolateral spur with one or two large teeth; intersegmental membrane III/IV and IV/V with two patches of coarse shagreen. Larva : mentum with sixteen teeth; median teeth ½ times shorter than first laterals; mandible with four inner teeth. Description. Male (n=2). Size . Total length 4.7–5.0 mm. Wing 2.0– 2.4 mm long and 0.6–0.7 mm wide. Coloration . Head yellowish; fourth and fifth palpomere and antenna brown. Thorax scutum yellowish brown with brown mesonotal stripes and short brown triangular spot before the scutellum; postnotum brown ( Figs 1 a, b). Abdomen brown as shown in figure 1c. Legs pale brown. Head. Temporal setae 20. Clypeus with 22–24 setae. Antenna 1.06–1.20 mm long; AR 1.4–2.0. Palpomere 2–5 lengths: 31–44, 175–219, 100–125, 118–150 µm. Thorax. Ac 21; Dc 14–19; Pa 5; Scts 13–16 coarse and 7–13 finer setae on anterior face.. Wings . VR 1.06. Brachiolum with two setae. Setae present on veins R with 28, R1 with 28 and R4+5 with 32 setae, remaining veins bare. Squama with 10 setae. Legs . Scale on fore tibia 38–46 µm long. Tibial combs of mid and hind legs partly fused; mid tibial spur anterior short 30 µm, posterior 69 µm long; hind tibial spur anterior 46–50 µm, posterior 81–84 µm long ( Fig. 1 e). Lengths and proportions as in Table 3 . TABLE 3. Lengths (in Μm) and proportions of legs of Endotribelos bicolor sp. n. male.
fe ti ta1 ta2 ta3
p1 1230–1277 1015–1077 1200–1292 760–954 569–769
p2 1107–1246 969–1108 584–692 323–400 230–292
p3 1230–1369 1046–1261 876–969 507–569 415–461
ta4 ta5 LR BR
p1 523–631 215–231 1.18–1.20 5.6
p2 153–169 66–123 0.60–0.62
p3 276–292 92–138 0.77–0.84
Hypopygium ( Fig. 1 e). Anal point 75–84 µm long; 9 µm wide medially; 12–15 µm near apex. Tergite IX with 15–21 strong setae. Laterosternite IX with two setae. Phallapodeme 112–133µm long. Gonocoxite 146–169 µm long. Superior volsella 123 µm long, broad basally and strongly curved distally, with 3–4 long basal setae and one 65–77 µm long sub-apical seta near 27 µm distal from extremity ( Fig. 1 f). Gonostylus 209–250 µm long. HR 0.67–0.69. HV 2.05–2.18. Female (n=1). Dimension and coloration. Wing length 2.2 mm . Coloration as male except the brown markings of the posterolateral corner of scutum. Head. Temporal setae 8, uniserial. Clypeus with 28 setae. Antenna 0.68 mm long; AR 0.36. Palpomere 2–5 lengths: 37, 188, 112, 144 µm Thorax. Ac 20; Dc 25; Pa 6; Scts 24. Wings . VR 1.13. Brachiolum with two setae. Setae present on veins R, R1, R4+5 and M, remaining veins bare. Squama with 10 setae. Legs . Scale on front tibia 47 µm long. Tibial combs of mid and hind legs partly fused; mid tibial spur 31 µm long; hind tibial spur anterior 53 µm, posterior 67 µm long. Lengths and proportions as in Table 4 . TABLE 4. Lengths (in Μm) and proportions of legs of Endotribelos bicolor sp. n. female.
fe ti ta1 ta2 ta3
p1 1338 1061 1492 953 707
p2 1292 1138 - - -
p3 1307 1261 984 584 446
ta4 ta5 LR BR
p1 615 184 1.11 5.3
p2 - - -
p3 292 123 0.75
Genitalia ( Fig. 2 ): Setal patches on sternite VIII with 17–18 setae. GC IX with two setae. Segment X with 8 setae. Seminal capsule 91 µm long. Notum 128 µm long, rami 66 µm. Coxosternapodeme IX 168 mm long. Cercus 106 µm long. Pupa (n=4, 2 males and 2 females ). Abdomen 4.2–5.1 mm long. Wing sheath 1.4–1.6 mm long. Exuviae coloration pale brown. Cephalothorax . Cephalic tubercles 109 µm long, conical, apically with several pointed spinules; frontal setae 144 µm long ( Fig. 3 a). Antepronotal seta 1, precorneal setae 2, dorsocentral setae 4. Distance between Dc1 and Dc2 44–75 µm, between Dc2 and Dc3 198–316 µm, between Dc 3 and Dc 4 22–34 µm. FIGURE 3 . Endotribelos bicolor sp. n. Pupa. a . cephalic tubercle. b, c. Abdominal tergites I-IV and detail of hook spines. d . Posterolateral spur of segment VIII. FIGURE 4 . Endotribelos bicolor sp. n. Larva. a . Dorsal sclerites of head. b . Pecten epipharyngis. c. Antenna. d . Mandible. e. Mentum and ventromental plate. Abdomen. Tergite I bare, II–VI covered with shagreen, spines near central part of anterior margin stronger, VII and VIII bare. Paratergites V and VI with posterior patches of fine spines. Conjuntives III/IV and IV/V with two large patches of anteriorly directed large spines ( Fig. 3 b). Hook row continuous with near 32 large curved spines occupying near 1/3 width of segment II. Pedes spurii A and B present. Spur on S VIII with one large apical tooth and 0–1 lateral tooth ( Fig. 3 c). Abdominal lateral setation: segment II–IV with three L setae, V–VI with three taeniate setae, VII–VIII with four taeniate setae. Anal lobes with 32–40 taeniate fringe setae on each side. 4th instar larva . Head capsule yellowish with mandibular tips, mentum and occipital ring blackish. Head . Dorsal head length 625 µm; width 562 µm; IC=0.89. Antennal basal segment 77 µm long. AR 1.02; antennal blade 61 µm long, shorter than antennal flagellum ( Fig. 4 c). Dorsal sclerites of head as in figure 4a, clypeus 62 µm long. Pecten epipharyngis formed by three toothed plates ( Fig. 4 b). Premandible with two major and one minor teeth; brush well developed. Mandible 320 µm long, with small dorsal tooth, apical and four inner teeth dark brown; seta subdentalis slender, 61 µm long ( Fig. 4 d). Mentum 215 µm wide, with sixteen dark teeth; median teeth 1/2 shorter than first laterals; 2nd lateral teeth smaller than the 3rd lateral teeth ( Fig. 4 e). Ventral plate 143 µm long, with very distinct striations, distance between ventramental plates 69 µm.