A review of the Japanese species of the family Tischeriidae (Lepidoptera)

Abstract This paper provides taxonomic and biological data on one new and one newly recorded species of Coptotriche Walsingham and one new and one newly recorded species of Tischeria Zeller from Japan. Coptotriche symplocosella Kobayashi & Hirowatari, sp. n. (host Symplocos lucida, Symplocaceae), and Tischeria kumatai Sato, Kobayashi & Hirowatari, sp. n. (host Tilia japonica, Malvaceae) are described. The pupal morphology of Coptotriche symplocosella is illustrated with scanning electron micrographs. Coptotriche minuta Diškus & Stonis, 2014 and Tischeria relictana Ermolaev, 1986 are newly recorded from Japan. The female, hostplants (Carpinus, Corylus, and Ostrya species), and immature stages of Coptotriche minuta and the adult features, female, and hostplants (Betula species) of Tischeria relictana are described with photographs and drawings for the first time. Mine types and characters of Japanese Tischeriidae are reviewed with photographs.


Introduction
The Tischeriidae is a lepidopteran family comprising some of the smallest moths, with a wing expanse of only 5-11 mm. Tischeriid adults are rather similar to one another in appearance, with a brown or blackish gray vestiture. The family can be distinguished from other families by a frontal tuft projecting over a triangular face smoothly covered with scales, numerous, long and recurved, cilia-like sensilla trichodea (see Davis 1986 and van Nieukerken and Dop 1987) on the male antenna, in the male genitalia by a strongly narrowed phallus, usually bifurcate or with apical spines, and in the female genitalia with four to five pairs of apophyses (Puplesis and Diškus 2003). Puplesis and Diškus (2003) recognized three genera: Tischeria Zeller, 1839, Coptotriche Walsingham, 1890, and Astrotischeria Puplesis & Diškus, 2003. Until now, 115 tischeriid species have been described globally. Nearly eleven plant families have been reported as their hosts, among which Rosaceae, Fagaceae and Asteraceae are major groups (Puplesis and Diškus 2003;Stonis et al. 2014). Several species of Tischeriidae have recently been added to the Eastern Palaearctic fauna, including two species from China (Huang and Tan 2009) and two new species from the Russian Far East (Stonis et al. 2014).
In Japan, two genera and seven species of Tischeriidae have been described to date (Sato 2011; see checklist below). In addition, some unnamed species have also been collected (Oku 2003, Sato 2011. Among them, according to Sato (2011), two are unidentified Tischeria species associated with Tilia (Malvaceae) and Betula (Betulaceae) respectively, and two are unidentified Coptotriche species associated with Carpinus (Betulaceae) and Quercus (Fagaceae) respectively.
In this paper, we taxonomically review the Japanese species of the family Tischeriidae, resolving the identity of three of these unidentified species with descriptions of two new species and two newly recorded species. For the two Coptotriche species, larval and/or pupal stages are also described. Eight Japanese species were reared, and their mine types and characters are reviewed with photographs.

Materials and methods
Adults were collected with light traps and leaves with mining larvae and cocoons were sampled from March to November in 2008 to 2015 in locations shown in Table 1. Adult specimens are preserved in the Osaka Prefecture University (OPU) and Tokushima Prefectural Museum (TKPM). Immatures in leaves were reared in plastic cups (420 ml: 129 mm in top diameter and 60 mm in depth) containing wet cotton at 20±5 ˚C under a photoperiod of 13-16L : 8-12D in the laboratory. In addition, specimens collected by Dr H. Kuroko in OPU, those collected by Dr T. Kumata in Hokkaido University Museum (HUM), and collections of the third author (Hirano) were examined.
Photographs of leaf mines were taken primarily in the field using an OLYMPUS μ1060 digital camera. Some leafmines were scanned using an EPSON GT7400. Some pupae were dried and sputter-coated with a 60 : 40 mixture of gold-palladium for examination with a scanning electron microscope (SEM). SEM photographs were taken using HITACHI SU1510 with a lanthanum hexaboride (LaB6) source at an accelerating voltage of 15 kV. For preparation of the male and female genitalia, the abdomen was removed and boiled for 3-4 min in 10% aqueous KOH. They were stained with acetocarmine.
Terms for genitalia, in principle, follow Stonis et al. (2014). The term "prela" introduced by Braun (1972) is used to designate the two or three paired rod-like or plate-like apophyses that extend from the inner side of the 8 th and 9 th sternites. The term "antrum" is employed to indicate the strongly thickened, differently shaped walls of the vestibulum following Puplesis and Diškus (2003). Scientific names of plants follow the Missouri Botanical Garden Tropicos database (2015).
Male genitalia ( Remarks. Two pairs of prelae were observed to expand caudally and form a humpshape in the female genitalia of some specimens ( Fig. 3Db-d).
The folded, leaf-edge mines of this species resemble at first sight those of foreign congeneric species feeding on Fagaceae and Rosaceae, e.g. Coptotriche citrinipennella (Clemens, 1859), C. gaunacella (Duponchel, 1843), C. crataegifoliae (Braun, 1972) and C. agrimoniella (Braun, 1972). However, larvae of C. citrinipennella form more tightly folded and narrow mines (Braun, 1972), while other species form more expanded mines. Fully expanded mines of C. minuta are easily distinguishable from those of the other Japanese Coptotriche species as shown in Fig. 13, although all of them are irregular blotch mines lined with a few folds. The mine of C. minuta is most similar to that of C. angusticollella, but the fold of C. minuta is obviously smaller than that of C. angusticollella. A mine of C. minuta may look like a pupal shelter of Roeslerstammia pronubella ([Denis & Schiffermüller], 1775), Roeslerstammiidae, which utilizes the same hostplant, Carpinus laxiflora (Hirowatari et al. 2012, figs 7, 8 Diagnosis. The color of the scaling is very similar to that of many other Coptotriche species; the new species differs from other members of the genus in the combination of the rather long uncus (Fig. 3F), and the gently curved slender valva (Fig. 3G) in the male genitalia, and the very small corpus bursae (Fig. 3H) in the female genitalia.
Pupa. (Fig. 11) (3 preparations examined). Brown to dark brown, 4.4-5.1 mm in length. Vertex (Fig. 11A-B) smooth, with a pair of short setae laterally (Fig. 11C). Dorsum A2-A7 with a pair of long setae, and a concentration of very small spines (Fig.  11E). Dorsum A8-A10 (Fig. 11G, H)  Biology (Figs 9-10). Because many young larvae were observed in leaf mines in March, the species seems to overwinter in the larval stage. The larvae mine leaves of an evergreen tree, Symplocos lucida, forming an elongate full-depth blotch mine beginning with a slender, linear shape (Fig. 9A, E), and gradually expanding as they feed and grow (Fig. 9B, F, G); about ~3 cm in length, white to dark yellow; the older mines turn brown in coloration (Fig. 9C). There were usually 1-3 mines per leaf (Fig. 9G). The larva ejects frass through circular holes (Fig. 10). From shed larval head capsules in the mine, we estimate that the species has six larval instars (Figs 9I, 10). The semifinal and final instar larvae are 6.0-7.0 mm long and pale green in coloration. Head capsule widths are 1st instar: 0.21 mm, 2nd: 0.25 mm, 3rd: 0.30 mm, 4th: 0.40 mm. The mature larva lines the mine with silk, so that the upper surface of the mine shows a few folds (Fig. 9C); a pupal cocoon is situated at the end of the mine.
Remarks. The pupal characters of the new species are similar to those of other Coptotriche species, but the new species has rather short caudal processes. Diagnosis. The color of the scaling of this new species has little or no diagnostic value. However, the female genitalia exhibit good diagnostic characters, especially the thickened plate-like vestibulum (antrum) (Fig. 4I). Among Tischeria species having a similarly unusual antrum (e.g. T. ptarmica Meyrick (see van Nieukerken 2010) and T. zestica Meyrick (see Puplesis and Diškus 2003), the new species is most similar to T. unca Diškus & Stonis from the Russian Far East (feeding on Quercus), but is recognizable by the slender posterior plate of the antrum and the lack of spines in the corpus bursae in the female genitalia, and the long spiral shaped juxta (Fig. 4E-F) and the valva with a very slender basal half (Fig. 4A) in the male genitalia. The South African species, T. antilope Puplesis, Diškus & Mey (female unknown) also has a similarly shaped juxta and valva, but differs from the new species by the narrow ventral plate of the vinculum, the longer valva and the lack of a pair of short lateral processes on the juxta (Puplesis and Diškus 2003, figs 586, 589). A Far Eastern Russian species, T. puplesisi Kozlov (female unknown), differs from the new species by the broader valva and short, stout juxta (Kozlov 1986 , fig. 2). Adult (Fig. 2F-G). Wing expanse 7.3 mm; forewing length 3.5 mm in holotype, 7.4 and 8.4 mm (3.3 and 3.9 mm) in paratypes. Head: palpi cream white; face smooth, cream white; vertex tuft white mixed with slender pale ocherous lamellar scales; antenna slightly longer than or equal to half of forewing, basally pale ocherous, apically ocherous to brown. Thorax pale ocherous. Forewing pale ocherous with scattered brown scales on the costal half towards the apical area. Cilia and hindwing blackish gray. Legs white to pale ocherous. Abdomen: brown; anal tuft grayish ocherous.

Tischeria kumatai
Male genitalia (Fig. 4A-G) (1 preparation examined). Uncus with very slender lateral lobes (Fig. 4C). Socii membranous. Tegumen narrow, marginally reinforced with a pair of slightly inwardly curved arms (Fig. 4B). Valva long, basally very slender, apically rounded, distally covered with fine setae and with a long slender dorsal process (Fig. 4A). Ventral plate of vinculum long and narrow, rounded anteriorly. Juxta very long, comprising two pairs of spiral curved processes (Fig. 4D-F); one pair connecting to the middle of the phallus, equal to the length of valva, apically forming a spiral shape (Fig. 4E-F); the other pair more slender, half the length of the valva, basally slightly curved laterally (Fig. 4D). Phallus slender, distinctly broadened at basal end, forming a pale slender membranous structure from the middle to apex.

Host plants. Tilia japonica (Miq.) Simonk. (Malvaceae).
Etymology. The specific epithet, kumatai, is dedicated to Dr Tosio Kumata, who is one of the great Lepidoptera taxonomists and collected the holotype and some of the paratypes.
Biology. The larvae form dark gray blotch mines on the leaf edge which are very similar to the folded mines of Coptotriche minuta on Carpinus japonica. Tischeria kumatai is common in Nagano Prefecture. Ermolaev, 1986 Figs 2H-L, 5 Tischeria relictana Ermolaev, 1986: 6-8, fig  Host Betula grossa, N. Hirano leg.: 1♂, Oshirakawa, Azumi, Matsumoto, Nagano Pref., 25.iv.1990em., 23.x.19891♀, same locality, 20.v.2004em., 11.x.2003 Diagnosis. Tischeria relictana resembles Coptotriche species associated with Rosaceae in that the wings and thorax are covered with gray scales. However, this species can be regarded as a member of Tischeria by the presence of a developed juxta in the male genitalia. Although having divided valvae as well as some other congeneric species (e.g., T. zestica Meyrick and T. martinkrugeri Puplesis & Diškus), T. relictana clearly differs from the others in the double juxta with anteriorly semicircular sclerotized diaphragma (Fig. 5A). A Far Eastern Russian species, T. sichotensis Ermolaev, has female genitalia similar in shape to T. relictana, but the former is separated from the latter by the presence of two acute lateral lobes of the antrum and the short spine-like pectinations in the caudal part of the corpus bursae (Stonis et al. 2014, figs 42, 43). (Fig. 2H-L). Male and female. Wing expanse 6.1-7.1 mm; forewing length 3.0-3.2 mm in Japanese specimens. Head: palpi blackish brown; face smooth, blackish brown; vertex tuft blackish brown; antenna blackish brown, equal to half of forewing in length. Thorax black. Forewing blackish brown to black. Cilia and hindwing blackish gray. Legs blackish brown. Abdomen blackish brown; anal tuft grayish ocherous.

Additional description. Adult
Male genitalia (Fig. 5A, B) (2 preparations examined). Uncus with long and very slender lateral lobes. Socii membranous. Tegumen strongly sclerotized marginally with a pair of slightly inwardly curved frames. Valva (Fig. 5B) broad, covered distally with fine setae, and having a long, slender, dorsal process. Ventral plate of vinculum narrow, triangular. Juxta short, comprising two pairs of processes, one pair connecting to the middle of the phallus, half the length of the valva, narrow medially, broadened apically; the other pair needle-shaped, 1/4 length of valva, slightly broadened basally. Transtilla absent. Diaphragma anteriorly sclerotized, a semicircular plate, folding round the phallus and contacting the needle-shaped part of the juxta ventrally. Phallus (Fig. 5A) slender, distinctly broadened at basal end, forming a pale slender membranous structure from the middle to apex.
Female genitalia (Fig. 5C, D) (3 preparations examined). Similar to T. sparmanniae and T. zestica, but differs in having short apophyses anteriores and posteriores, a slender ductus bursae and the corpus bursae without spines.
Host plants. Betula ermanii Cham., B. grossa Siebold & Zucc. (Betulaceae). Biology. The detailed biology of this species is unknown. The larvae mine leaves of Betula spp., according to label data of adult specimens.
Biology. (Fig. 13-3). See Sato (1993) and Stonis et al. (2014). Remarks. In Japan, this species had been treated as 'T. decidua Wocke' until the East Asiatic subspecies T. decidua siorkionla was described by Kozlov (1986). Stonis et al. (2014) reported that Japanese representatives of T. decidua belonged to the subspecies siorkionla Kozlov. In the Japanese specimens we studied, the apex of the valva is broader (present study: 50-65 μm; Stonis et al. (2014): 65 μm), but other characters were considered to lie within the range of individual variation, e.g., some specimens have a rather prominent median bulge and sinuous inner margin of the valva (Fig.  6A, D, G, L; Sato 1993, 4D), i.e. more similar to that of the nominotypical European subspecies; others have a rather longer but less prominent median bulge and nearly straight inner margin of the valva (Fig. 6J, K, M; Sato 1993, fig. 4A, E), i.e. more similar to that of T. d. siorkionla. The chitinized basal part of the phallus tends to be less developed and the transverse bar is shorter in Japanese material than in the nominotypical European subspecies (Fig. 6C, F, I), as shown by Stonis et al. (2014).

Discussion
In the present study, a total of eleven Tischeriidae species are recognized from Japan, not including an unidentified Tischeria species which occurs on evergreen Quercus (recorded by Sato 2011), of which nine of them were reared by us. It is revealed that the previously unknown hostplants for Coptotriche minuta were Carpinus spp., Corylus sieboldiana and Ostrya japonica, Betulaceae, while those for Tischeria relictana were Betula spp, Betulaceae. Coptotriche symplocosella and C. japoniella utilize evergreen plants of Ericales as hosts and hibernate in the larval stage. Oishi and Sato (2009) reviewed the voltinism and leaf type of hostplants of the Tischeriidae; C. japoniella has a univoltine life cycle and long larval period, and overwinters as 5 th instar. The seasonal development of C. symplocosella was not examined, but it has a similar hibernating form and larval mine as C. japoniella. Other Japanese species including Tischeria kumatai and C. minuta probably have a bivoltine life cycle and hibernate as mature larva within the cocoon. The overwintering generation of C. minuta has color morphs of forewings from ocherous to black and a larger body size than other generations. According to Kuroko (1982), Coptotriche szoecsi japonica has a brighter color and smaller body size compared with the nominotypical subspecies szoecsi. The subspecies japonica was described from specimens collected in May and July. The polytypic concept of C. szoecsi could not be confirmed, because there existed no opportunity to examine additional material. Figures 12 and 13 provide a pictorial key to the leafmines of Japanese Tischeriidae. Mines of Coptotriche species are distinguished from those of blotch miners of other lepidopteran families by the ejection of frass through holes and the folds of the fully expanded mine; e.g., C. japoniella (Fig. 13(8ab)) is distinguished by these characters from the Eurya blotch miner, Lyonetia euryella Kuroko, 1964. According to Sato (1993), Japanese Tischeria mines are distinguished mainly by colors and patterns of the surface and cocoon nidus ( Fig. 13(1-3)).
As regards the morphology of the female genitalia, Puplesis and Diškus (2003) pointed out that the corpus bursae and ductus spermathecae have great value in generic and species diagnoses. Coptotriche species were often distinguished from one another by the size of the corpus bursae and the length of the ductus spermathecae. Virgin female adults of five Japanese Coptotriche species obtained from rearing in the laboratory were examined, not including C. szoecsi (Kasy). The size of the corpus bursae and the length of the ductus spermathecae differed from one another as indicated in the species descriptions, e.g., the corpus bursae of C. angusticollella was larger than in the other four species.
Biodiversity Center, Netherland) and Dr D. R. Davis (Smithsonian Institution, USA) for their critical comments and improvement of the manuscript. Dr J. R. Stonis (Lithuanian University of Educational Sciences, Lithuania) provided us with important literature and Sir Anthony Galsworthy corrected the language of the manuscript. We also thank Ass. Prof A. Nakamura (Laboratory of Landscape Architecture and Conservation, OPU) for identifying hostplants. This research was partly supported by the Research Fellowships of the Japan Society for the Promotion of Science (JSPS) for Young Scientists (DC1, 1081000108), and JSPS and NSFC under the Japan-China Scientific Cooperation Program.
The first author wishes to express cordial thanks to Prof M. Ishii and Ass. Prof N. Hirai (Entomological Laboratory, OPU) for their valuable suggestions. He thanks the members of the Entomological Laboratory (OPU) for their kind advice and help. He also thanks Mr Y. Kobayashi (Soni, Nara) and Mr S. Yamaguchi (Nabari, Mie), Mrs M. Kobayashi (Soni, Nara), Mrs N. & H. Sugimoto and T. Miyata (Nabari, Mie), and his family for providing habitat information on host plants and/or facilities for his fieldwork.