Phylogeny of marine Ainuyusurika tuberculata (Tokunaga) (Diptera: Chironomidae: Chironominae), with description of the immature stages Author Han, Wu Author Tang, Hongqu text Zootaxa 2019 2019-11-07 4695 2 131 147 journal article 24945 10.11646/zootaxa.4695.2.3 1f8b6014-8a6a-4376-8376-2b43cf5c9825 1175-5326 3531250 68BA1999-5EC9-44C8-B007-0E4C83DF43EE Ainuyusurika tuberculata (Tokunaga) ( Figs. 2–5 ) Polypedilum ( Pentapedilum ) tuberculatum Tokunaga, 1940: 290 . Ainuyusurika tuberculata : Sasa 1988: 80 ; Oyewo & Saether 2008: 138 ; Yamamoto & Yamamoto 2015: 109 . Pentapedilum yakuabeum Sasa et Suzuki, 2000: 56 . Polypedilum ( Polypedilum ) yakucedeum Sasa et Suzuki, 2000: 57 . Ainuyusurika yakuabeum : Oyewo & Saether 2008: 139 . Ainuyusurika yakucedeum : Oyewo & Saether 2008: 140 . Material examined. Pe, China : Guangdong Province , Shantou City, Nan’ao County, Shen’ao Bay, Tayu Island, 5.vi.2016 , H.Q. Tang ; Pe , as previous except Tsing’ao Bay , 27.viii.2016 ; 4 males , 2 females , 4 Pe , 5 L, as previous except Treasure Island , 23°28’42.7”N , 117°07’22.9”E , 24.iii.2018 (emerged 5–8.v.2018 ) , H.Q. Tang; 4 male , 6 Pe , as previous except 21.iv.2018 ; 2 males as previous except 15.vi.2019 ; 1 male , China : Zhejiang Province , Wenzhou City , Dongtou Distinct , Dong’ao village , 4.v.2016 , H.Q. Tang; 10 male , Sourth Korea, South Jeolla , Sinan County , Jeungdo Island , 35°00’04.61”N 126°07’52.01”E , 8.viii.2019 , H.Q. Tang. Description. Male (n = 6). Total length 3.0–3.4, 3.2 mm . Wing length 1.4–1.9, 1.7 mm . Coloration. Generally pale brown, with dark brown vittae, lateral stripes on pleurae, postnotum and several intersegmental sections ( Fig. 2 A–B) Head. AR 0.95–1.19, 1.14. Ultimate flagellomere 420–510, 470 μm long. Wing ( Fig. 2C ). VR 1.29–1.36, 1.32. R with 14–19, 17 setae; R 1 with 15–22, 18 setae; R 4+5 with 12–18, 15 setae; M 1+2 with 5–18, 13 setae; M 3+4 and other veins bare. Wing cell r 4+5 with 17–33 macrotrichia; m 1+2 and m 3+4 with sparse macrotrichia; other cells bare. Squama with 6–10, 8 setae. Legs ( Figs. 2 E–H). LR 1 1.15–1.23, 1.19; LR 2 0.5–0.56, 0.52; LR 3 0.69–0.72, 0.71. BV 1 1.7–1.98, 1.83; BV 2 3.58–3.78, 3.65; BV 3 2.63 –2.9; 2.78. SV 1 1.75–1.86, 1.8; SV 1 3.92–4.5, 4.27; SV 1 2,89–3.26, 3.03. BR 1 1.2–2.25, 1.76; BR 2 1.5–3, 2.05; BR 3 2.4–4.2, 3.03. FIGURE 2 . Ainuyusurika tuberculata , male. A, Lateral view; B, dorsal view; C, wing; D, hypopygium. E, fore tibia. F, mid tibia. G–H, hind tibia the red arrow indicating the narrow gap (scale bar: A, B, C, 200 μm; D–H, 20 μm). FIGURE 3 . Ainuyusurika tuberculata , female. A, Lateral view; B, apex of wing; C, antenna; D, genitalia, ventral view; F, genitalia, dorsal view (scale bar: A–B, 200 μm; C, 60 μm; D-E 50 μm). FIGURE 4 . Ainuyusurika tuberculata , pupa. A, Cephalothorax; B, thoracic horn; C, thorax, lateral view; D, abdomen, dorsal view; E, anal combs of abdominal segment VIII (scale bar: D, 300 μm; others, 100 μm). FIGURE 5 . Ainuyusurika tuberculata , larva. A, Dorsal sclerites of head capsule; B, antenna; C, premandible; D, mandible; E, mentum (scale bar: A–D, 25 μm; E, 50 μm). FIGURE 6 . Color photos of Ainuyusurika tuberculata , pupa (A–D) and larva (E–I). A, Pharate female, lateral view; B, pupal exuviae; C, frontal apotome; D, anal comb of abdominal segment VIII; E, habitus of larva, lateral view; F, procercus; G, head, ventral view; H, SI setae; I, ventromental plate (Scale bar, A, B, E, 200 μm; C, D, F, G, 30 μm; H–I, 5 μm). Hypopygium ( Fig. 2D ). HR 1.06–1.3, 1.15; HV 2–2.68, 2.34. Female (n = 5). Total length 2.7–3.0, 2.8 mm . Wing length 1.6–2.2, 2.0 mm. Head . AR 0.38–0.46, 0.41. Ultimate flagellomere 128–155, 138 μm long. Wing ( Fig. 3B ). VR 1.35–1.39, 1.37. R with 16–20, 18 setae; R 1 with 23–24, 24 setae; R 4+5 with 24–31, 28 setae; M 1+2 with 22–45, 36 setae; M 3+4 with 12–20, 17 setae; other veins bare. Number of macrotrichia 45–90, 73 on wing cell r 4+5 ; 18–35, 25 on m 1+2 ; 5–11 on m 3+4 ; other cells bare. Legs . LR 1 1.18–1.23, 1.11; LR 2 0.48–0.50, 0.49; LR 3 0.53–0.68, 0.62. BV 1 2.05–2.13, 2.0; BV 2 3.92–4.3, 4.1; BV 3 3.02–4.43; 3.52. SV 1 1.92–1.93, 1.92; SV 2 4.10–4.32, 4.22; SV 3 3.03–4.04, 3.42. Genitalia (Figs. D–E). Gonocoxite IX small, with 2–3 lateral setae. Tergite IX broad, undivided, with 8–10 setae on each side. Pupa (n = 4). Total length 3.0–4.0, 3.5 mm . Coloration : Generally pale yellow except for light brown apophyses and anal combs ( Fig. 6A ). Cephalothorax ( Figs. 4A , 6C ). Cephalic tubercle 35–50, 43 μm wide at base. Frontal seta 30–45, 38 μm long; distance between two frontal setae 100–250, 121μm long ( Fig. 4A ). Thorax and thoracic horn as Fig. 4 B–C. Abdomen ( Fig. 4D ). A II with transverse row of 30–42 hooks, covering 60–80% of segment width. largest tooth of anal comb about 30 μm ( Figs. 4E , 6D ). Anal lobe with fringe of 10–14 taeniae; dorsal seta absent. Male genital sac distinctly beyond distal tip of anal lobe. Fourth instar larva (n = 5). Total length 4.5–5.0, 4.7 mm . Head ( Fig. 6G ). Dorsal sclerites as Fig. 5A . SI as Fig. 6H . Head 210–300, 250 μm long, cephalic index 0.73– 0.88, 0.83. Lengths of first to fifth antennal segments (in µm) ( Fig. 5B ): 40–50, 47; 8.3–12.5, 10.7; 4.8–5, 4.9; 6.6–8, 7.6; 3.2–5.2; 4.2; AR 1.1–2.0, 1.5. Basal segments 3.3–4.8, 3.8 times as long as basal width. Blade 23–35, 38 μm long; Bl/Fl 0.8–1.0, 0.92. Premandible ( Fig. 5C ) 42–60, 53 μm long. Mandible ( Fig. 5D ) 50–60 µm long.. Mentum 50–73, 63 µm wide, with 4 median and 6 pairs of lateral teeth ( Fig. 5E ). Ventromental plate 50–82, 70 µm wide ( Fig. 6I ). Distance between setae submenti 100–120, 106 µm long. Postmentum 167–230, 206 µm long. Body ( Figs. 6 E–F). Procercus 30–40 µm high, slightly longer than basal width, with 7–8 anal setae. Anal setae 220–250, 233 µm long. Supraanal seta 60–80, 75 µm long. Molecular analysis. Ten COI-5P fragments of adults, larvae and pupae submitted to the BOLD system showed that they matched three nominal A. tuberculata specimens, which formed three independent BINS. The maximum k2p intraspecific distance (including species retrieved from database) were 4.2% between specimens labeled TL0505L and AB704943 . The Neighbor-Joining (N-J) tree ( Fig. 7A ) based on available standard DNA barcodes of Ainuyusurika showed that Chinese population could be separated from Japanese population and South Korea population. The MrBayes (MB) tree ( Fig. 7B ) showed that Ainuyusurika was sister to a group comprising of Endochironomus , Endotribelos , Phaenopsectra and Sergentia . The whole multi-genera assemblage was sister to another group comprising of Endochironomus , Tribelos , and Synendotendipes . Endochironomus was demonstrated as a polyphyletic genus in this study, while the monophyly of other genera is uncertain due to limitations in available sampling of taxonomic diversity. Distribution . This species has been recorded from Sakhalin Island in Russia ( Tokunaga 1940 ); Hokkaido Island, Kii Penisula, Seto Inland Sea, Yakushima Island and Okinawa Islands in Japan ( Sasa 1988 ; Sasa & Suzuki 2000 ; Kawai et al . 2011 , 2015 ; Yamamoto et al . 2015 ); Gangwon-do in South Korea ( Ree 2013 ). We could add the records from Guangdong and Zhejiang in China and South Jeolla ( Jeollanam-do ) in Sourth Korea ( Fig. 1 ). Ecology. We conducted a year-long survey in the Nan’ao County, a small island located at the extreme edge of Guangdong Province in China during 2016. This island is isolated from the mainland and surrounded by the South Sea. The sea surface temperature ranges from 18.8°C (February) to 28.8°C (June and July). Salinity ranges 29.2–34.4‰. Ainuyusurika adults emerged from the end of spring to the earlier autumn (May to October) when the water temperature exceeded 23°C. The emergence was most abundant in July and August.