New record of Epermenia (Calotripis) sinjovi Gaedike (Lepidoptera: Epermeniidae) in China: DNA barcode, adult, immature stages, host plant and biology Author Wang, Yehao 0000-0002-4638-974X Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Shandong Normal University, Jinan 250014, China & 201813010204 @ stu. sdnu. edu. cn; https: // orcid. org / 0000 - 0002 - 4638 - 974 X 201813010204@stu.sdnu.edu.cn Author Teng, Kaijian 0000-0001-6772-9419 Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Shandong Normal University, Jinan 250014, China & kaijianjn @ 163. com; https: // orcid. org / 0000 - 0001 - 6772 - 9419 Author Liu, Tengteng 0000-0002-0182-2562 Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Shandong Normal University, Jinan 250014, China & liutt @ sdnu. edu. cn; https: // orcid. org / 0000 - 0002 - 0182 - 2562 liutt@sdnu.edu.cn text Zootaxa 2021 2021-09-15 5039 2 263 276 journal article 10.11646/zootaxa.5039.2.7 1175-5326 5509153 5A0C2BCF-5C85-4055-AFDE-1DF52BA41B04 Epermenia ( Calotripis ) sinjovi Gaedike, 1993 Epermenia ( Calotripis ) sinjovi Gaedike, 1993: 99 ; Budashkin 1996: 12 ; Budashkin 1997: 483 ; Budashkin & Gaedike 2005: 127 ; Kuroko & Gaedike 2006: 61 ; Sinev 2016: 149 ; Budashkin & Sinev 2019: 119 . TD: ZIN ( holotype , ♂ ; 33 paratypes , 16♂ , 17♀ ). TL: Primorskij Kraj , Okr. Diagnosis. Epermenia ( Calotripis ) sinjovi is similar to E. ( C .) chaerophyllella ( Goeze, 1783 ) superficially, but it can be separated from the latter by the aedeagus with one cornutus and the vesica bearing weakly sclerotized folds in the male genitalia ( Figs 13–15 ) and the ductus bursae with dense tiny coniform granules ( Fig. 18 ) and the places around signum with numerous scale-shaped granules ( Fig. 19 ) in female genitalia. In E . ( C .) chaerophyllella , the aedeagus has two cornuti and the vesica is inconspicuous and the ductus bursae bears thumbtack-shaped spinules and lacks scale-shaped granules ( Gaedike 1966a : Figs 87–88). Epermenia ( C .) sinjovi is also similar to E . ( C .) albapunctella Busck, 1908 and E . ( C .) californica Gaedike, 1977 on the female genitalia, but it can be distinguished from the latter two by the ductus bursae inflated towards lateral side and by the diagnostic characters used for E . chaerophyllella . Material examined. Adult. 6♂ , 1♀ , China : Shandong , Qingdao , Laoshan mountains , 36.204°N , 120.609°E , 400 m , larva coll. 01.vii.2017 , mine on leaves of Angelica polymorpha , emerged 15.vii.2017 (indoors), leg. Tengteng Liu and Zhenquan Gao , registered nos SDNU.QD170721.1 (genitalia slide no. WYH0011), SDNU.QD170721.2 (WYH0017), SDNU.QD170721.5 (WYH0014 ♀ ), SDNU.QD170721.6 (WYH0012), SDNU.QD170721.7 (WYH0013), DNA sequences successfully obtained from above specimens; SDNU.QD170721.3 (WYH0009), SDNU.QD170721.4 (WYH0016). Pupa . 5♂ (3 emerged), 1♀ , other data same as adult, deposited in 95% ethanol. Mine. Photos of two damaged leaves, other data same as adult. DNA barcode. Five DNA barcodes from individuals of E . ( C .) sinjovi were newly obtained and have been made public in BOLD ( Table 2 ). A neighbor-joining tree together with available DNA barcodes of other Epermenia spp. is given in Fig. 1 . Epermenia ( C .) albapunctella and E . ( C .) chaerophyllella are closest to E . ( C .) sinjovi , and all the new DNA barcodes of E . ( C .) sinjovi are grouped into a cluster. Adult ( Figs 2–5 ). Wingspan 9.5–11.4 mm . Head grayish fuscous to blackish fuscous, with scales around eyes grayish yellow basally. Antenna about 2/3 length of forewing; scape dilated, blackish fuscous dorsally, sometimes mixed with yellowish-brown scales, grayish white or yellowish brown ventrally; pectens yellowish gray except brown subapically; flagellum pale grayish brown with black annulations, except second to fifth flagellomeres black. Labial palpus grayish white on inner surface, with dense blackish fuscous scales which tipped with grayish white on outer surface. Thorax and tegula grayish fuscous or blackish fuscous, usually intermixed with grayish yellow scales tipped with brown. Forewing broadly lanceolate, ground color ochreous red; humeral area blackish fuscous; costa with dense gray scales tipped with dark brown, a broad black transverse fascia ranging from basal 2/5 to 2/3, with inner margin extending obliquely to basal 1/3 of dorsum and outer margin extending to basal 2/3 of dorsum; dense brown scales along upper margin of cell and below CuP before transverse fascia; distal 1/3 of forewing scattered with brown or dark brown scales; sometimes two small black dots in middle of and distal part of cell, edged with white scales, with a white dot between them, sometimes absent; dorsum with 3 clusters of black scale teeth, grayish brown basally, ranging from basal 1/5 to 2/3, distal 1/3 with dense pale brown scales tipped with black, forming 2 black lines on tornus; cilia grayish brown, pale grayish yellow basally. Hindwing and cilia pale grayish brown, slightly darker towards apex. Legs grayish white dorsally, dark brown ventrally, some scales tipped with grayish white; tibiae with black annulations apically; each tarsomere with black annulation, grayish white apically. Male with a pair of pockets with androconial scales from first to third abdominal segments latero-ventrally ( Fig. 16 ). Piliform scale tufts dense and long around genitalia. Note. According to the coloration of the forewing, Kuroko & Gaedike (2006) recognized four forewing patterns: the normal phenotype, the black-bar phenotype, the brown phenotype and the dark phenotype. Compared with the normal phenotype, the dot within cell is connected with the distal dot by a black bar in the black-bar phenotype, and the forewing is entirely suffused with black scales in the brown phenotype and the dark phenotype ( Kuroko & Gaedike 2006 ). Specimens examined in this study belong to the normal ( Figs 2–3 ) and brown phenotypes ( Figs 4–5 ). FIGURE 1. Neighbor-joining tree estimated from DNA barcodes of E . sinjovi and available DNA barcodes of other species in Epermenia from BOLD, presented by using Kimura 2-Parameter model. TABLE 2 Specimens and barcodes used in the neighbor-joining tree analysis

Species name	BOLD sample ID	Collecting date	GenBank Accession	Country	Depository
Epermenia ( Calotripis ) sinjovi	SDNU.QD170721.1	2017-7-15	-	China	Shandong Normal University
E . ( C .) sinjovi	SDNU.QD170721.2	2017-7-15	-	China	Shandong Normal University
E . ( C .) sinjovi	SDNU.QD170721.5	2017-7-15	-	China	Shandong Normal University
E. ( C .) sinjovi	SDNU.QD170721.6	2017-7-15	-	China	Shandong Normal University
E. ( C .) sinjovi	SDNU.QD170721.7	2017-7-15	-	China	Shandong Normal University
E. ( C .) infracta	10BBCLP-3064	2010-06-23	KM550984	Canada	Centre for Biodiversity Genomics
E. ( C .) infracta	HLC-23326	2005-06-16	KT134027	Canada	Centre for Biodiversity Genomics
E. ( C .) aequidentella	TLMF Lep 13395	2013-09-05	-	Austria	Tiroler Landesmuseen
E. ( C .) aequidentella	TLMF Lep 05225	2011-08-01	-	Macedonia	Tiroler Landesmuseen
E. ( C .) chaerophyllella	MM00443	2006-05-06	-	Finland	University of Oulu, Zoological Museum
E. ( C .) chaerophyllella	NHMO-DAR-8192	2015-06-08	-	Norway	University of Oslo, Natural History Museum
E. ( C .) falcata	BIOUG03128-H11	2012-07-23	KM553833	Canada	Centre for Biodiversity Genomics
E. ( C .) falcata	BIOUG06600-H10	2012-07-14	KM541150	Canada	Centre for Biodiversity Genomics
E. ( C. ) illigerella	MM05213	2006-07-22	HM872717	Finland	University of Oulu, Zoological Museum
E. ( C .) illigerella	NHMO-DAR-13680	2017-07-09	-	Norway	Research Collection of Kai Berggren
E. ( C .) imperialella	BIOUG26649-D07	2014-06-22	MG469204	Canada	Centre for Biodiversity Genomics
E. ( C .) imperialella	BIOUG27081-G04	2014-06-22	MG470409	Canada	Centre for Biodiversity Genomics
E. ( C .) falciformis	MM03123	-	HM871833	Finland	University of Oulu, Zoological Museum
E. ( C .) falciformis	TLMF Lep 12501	1998-06-09	-	Austria	Tiroler Landesmuseen
E. ( C .) stolidota	08-JWB-0088	2007-08-16	-	United States	Smithsonian Institution
E. ( C .) stolidota	JWB-07-2003	2007-08-18	KF492335	United States	University of Maryland
E. ( C .) insecurella	TLMF Lep 13524	2013-05-06	-	Austria	Tiroler Landesmuseen

......continued on the next page TABLE 2. (Continued)

Species name	BOLD sample ID	Collecting date	GenBank Accession	Country	Depository
E. ( C .) insecurella	TLMF Lep 17736	2014-09-05	-	Austria	Research Collection of Peter Buchner
E. ( C .) albapunctella	BIOUG02730-B05	2010-04-04	KT132857	Canada	Centre for Biodiversity Genomics
E. ( C .) albapunctella	BARS_2016_26_222	2016-06-13	MG364745	Canada	Centre for Biodiversity Genomics
E. ( Epermenia ) scurella	TLMF Lep 12373	2013-06-29	-	Italy	Tiroler Landesmuseen
E. ( E. ) scurella	BC ZSM Lep 25223	2006-07-26	HM422138	Germany	Research Collection of Alfred Haslberger
E. ( E. ) ochreomaculella	TLMF Lep 17940	2015-06-18	-	Italy	Tiroler Landesmuseen
E. ( E. ) ochreomaculella	KBE 2018169	2018-06-02	-	Croatia	Research Collection of Kai Berggren
E. ( E. ) pontificella	KBE 2018167	2018-05-27	-	Croatia	Research Collection of Kai Berggren
E. ( E. ) pontificella	TLMF Lep 05224	2011-07-28	-	Macedonia	Tiroler Landesmuseen
E. ( Epermeniola ) commonella	gvc11119-1L	2009-01-26	-	Australia	Research Collection of Graeme V. Cocks
E. ( E. ) commonella	gvc16339-1L	2011-02-19	-	Australia	Centre for Biodiversity Genomics
E . ( Cataplectica ) iniquella	TLMF Lep 06665	2007-08-22	-	Austria	Research Collection of Peter Buchner
E . ( C. ) devotella	TLMF Lep 14358	2002-07-28	-	Austria	Tiroler Landesmuseen
E. ( C. ) profugella	MM23300	2009-01-01	-	Finland	Research Collection of Leo Sippola
E. ( C. ) profugella	NHMO-DAR-13633	2014-07-22	-	Norway	Research Collection of Kai Berggren

FIGURES 2–10. Adults and wing venations of Epermenia ( Calotripis ) sinjovi 2–3, normal phenotype, male; 4, brown phe- notype, female; 5, brown phenotype, male; 6, forewing venation, slide no. WYH0014; 7–8, hindwing venations, slide nos. 7, WYH0016, 8, WYH0014; 9, Rs and M 1 , M 2 and M 3 arising from the same point basally; 10, Rs and M 1 , M 2 and M 3 separated basally. FIGURES 11–19. Genitalia and pockets of Epermenia ( Calotripis ) sinjovi 11 and 13, male genitalia and aedeagus, genitalia slide no. WYH0013; 12, valva with shorter cucullus, genitalia slide no. WYH0017; 14–15, aedeagus of variable lengths, 15, vesica protruded, length of aedeagus referring to the distance between blue dots, that of cornutus between red dots, genitalia slide nos. 14, WYH0011, 15, WYH0017; 16, pockets with androconial scales, slide no. WYH0009; 17, female genitalia, WYH0014; 18, coniform granules in ductus bursae; 18, signum with teeth along edge. Venation ( Figs 6–10 ). Forewing with R 1 reaching costal 3/5, R 2 and R 3 almost parallel, R 4 and R 5 stalked basally, R 5 to termen, M 1 and M 2 nearly parallel, CuA 1 close to M 3 and far from CuA 2 basally, CuP vestigial, 1A+2A bifurcated at base; accessory cell present but inconspicuous. Hindwing with Sc+ R 1 reaching costal 4/5, Rs and M 1 as well as M 2 and M 3 arising from same point ( Fig. 9 ) or separated at base ( Fig. 10 ). Note. Rs and M 1 , M 2 and M 3 of the hindwing are separated from each other at base in Kuroko & Gaedike (2006) , but are sometimes originated from the same point in the examined specimens. Male genitalia ( Figs 11–15 ). Uncus slightly curved ventrad, about 3/5 length of valva, basal half nearly parallel-sided, gradually narrowing to point. Tegumen subtriangular. Ampulla broad basally, curved ventrad, sharply narrowing to a point, extending to distal 1/3 of cucullus, with a sclerotized band basally extending obliquely to base of cucullus. Cucullus 1/3–1/2 length of valva ( Figs 11–12 ), basal half with a weakly sclerotized longitudinal band connecting with former sclerotized band, distal half triangular, costal margin straight and ventral margin oblique, apex blunt. Sacculus heavily sclerotized, 1/2–2/3 length of valva ( Figs 11–12 ), ventral margin nearly straight, dorsal margin an arched ridge, distal part curved dorsad and extending beyond inner margin of cucullus, fused with substrate, not protruded terminally. A thin and straight sclerotized fold from base of sacculus to base of ampulla. Anellus cup-shaped, heavily sclerotized. Vinculum a narrow band, strongly sclerotized. Aedeagus stout, rounded basally, 1.2–1.6 times as long as cornutus ( Figs 13–15 ); cornutus pointed basally, widened to basal 1/5, then nearly parallel-sided to basal 3/5, distal 2/5 narrower, apex blunt; vesica with weakly sclerotized folds. FIGURES 20–23 Larva and biology of Epermenia ( Calotripis ) sinjovi 20, larva; 21–23, mines, arrows indicating early small mines. Note. The length of the aedeagus is variable ranging from 1.2 times as long as cornutus ( Gaedike 1993 ) to 1.2–1.6 times (the present study) and 1.7 times ( Kuroko & Gaedike 2006 ). Gaedike (1993 : Figs 17–18 ) illustrated two types of the cornutus: one is spindle-shaped, narrow basally and apically; the other is pointed basally, widest at middle, distal part spoon-shaped, blunt apically. The cornutus of the specimens examined in this study belong to the latter type . Gaedike (1993) didn’t describe the gnathos in the original description, but Kuroko & Gaedike (2006) suggested an inverted Y-shaped gnathos in the redescription. The male genitalia of all the specimens examined in this study lack the gnathos. Both Gaedike (1993) and Kuroko & Gaedike (2006) suggested a pointed projection at the apex of the dorsal margin of sacculus, but the structure is fused with and not protruded from the substrate. So, it is not proper to define this structure as a projection. Female genitalia ( Figs 17–19 ). Papillae analis setose and blunt apically. Apophyses posteriores not reaching posterior margin of VII sternum; apophyses anteriores with basal 2/5 bifurcated, reaching middle of VII sternum, nearly as long as apophyses posteriores. Ostium bursae concave, semicircular, at middle of and about 1/2 width of posterior margin of VII sternum, slightly sclerotized. Antrum heavily sclerotized. Ductus bursae inflated towards lateral side, with dense tiny coniform granules ( Fig. 18 ). Corpus bursae a broad bag, with numerous scale-shaped granules on internal surface around signum. Signum near middle of corpus bursae, comprising of a subtriangular basal plate and a subtriangular process arising from it, with numerous teeth along edge of process ( Fig. 19 ). FIGURES 24–30 Pupa of Epermenia ( Calotripis ) sinjovi 24, ventral view; 25, dorsal view; 26, lateral view; 27, subcircular hollow surrounded by two crescent sclerotizations; 28, genitalia orifice and anus, male; 29, genitalia orifices and anus, female; 30, curved and hooked setae on cremaster. Note . The basal plate of the signum is variable in shape, for example, round, oval and triangular ( Gaedike 1993 ; Kuroko & Gaedike 2006 ). Examined specimen in the present study shows clear teeth along the edge of the process of the signum which have not been described in previous papers. Larva (Fig. 20). Head black; thorax and abdomen translucent, milk white; pronotum sclerotized strongly. Setae black, pinaculum colorless and transparent. Pupa (Figs 24–30). Length: 4.0– 4.7 mm , width: 1.3–1.6 mm . Yellowish fuscous to blackish fuscous over development. Vertex and frons plump, eye-piece large, almost as wide as frons. Antenna from dorsal side of eye-piece to ventro-posterior edge of AVII . Labrum isosceles-trapezoid-shaped. Labial palpus and proboscis from posterior edge of labrum, former extending straightly to anterior edge of AI and latter extending obliquely and reaching middle of AIV. Foreleg and midleg originated from posterior edge of eye-piece, extending obliquely to middle ventrally, reaching posterior edge of AIII and posterior edge of AIV, respectively; femur of foreleg visible between proboscis and foreleg, extending obliquely to middle ventrally, narrow fusiform, nearly as long as labial palpus; hindtarsus originated from terminal of proboscis, extending straightly to posterior edge of AVII . Forewing extending obliquely to middle ventrally with costa along antenna, covering most lateral side of body; hindwing extending obliquely to middle ventrally, beneath forewing with only dorsum visible. TI – TIII sclerotized and wrinkled, with an oval bulge on middle of TII. AII– AVII with 1 spiracle dorso-laterally, with 1 seta dorsal to spiracle on AI– AIII , 2 setae dorsal and ventral to spiracle on AIV– AVII with strongly sclerotized annulus on posterior edge, respectively. AVIII – AX strongly sclerotized; AIX and AX fused ventrally, with a subcircular hollow laterally which surrounded by 2 crescent sclerotizations. (Fig. 27). Genital orifice of male on AIX (Fig. 28), those of female on AVIII and AIX (Fig. 29), anus on AX , genital orifice and anus linear, sclerotized strongly. Cremaster with 9–11 curved and hooked setae (Fig. 30). Distribution. China ( Shandong ), Japan ( Kuroko & Gaedike 2006 ), Russia ( Sinev 2016 ; Budashkin & Sinev 2019 ), Kunashir Is. ( Gaedike 1993 ). Host plants. Angelica polymorpha Maxim. (Apiaceae) in China , new record ; A . pubescens Maxim. and A . ursina Maxim. in Japan ( Kuroko & Gaedike 2006 ). Biology (Figs 21–23). Early instar larvae mine into leaves from hypodermis, and feed mesophyll surrounded by fine veins. Early instar mines are distributed irregularly on leaves. One larva can create new mines during larval stages. Early mines are linear and then expand to translucent patches. Mines at the base of leaflets increase over the growth of larvae, and gradually connect with each other, forming a large translucent patch and extending towards the distal end of the leaflet. Mines are white from upper-side view. Larvae spin a thin web on the underside of a leaf. Frass occur both in mines and attached on the web on the lower surface of a leaf, without the habit of cleaning frass. Larvae of different instars live together.