Agnė Rocienė TAXONOMY of the NEPTICULIDAE OF

LITHUANIAN UNIVERSITY OF EDUCATIONAL SCIENCES

Agnė Rocienė

TAXONOMY OF THE NEPTICULIDAE OF CONTINENTAL EAST ASIA (INSECTA, LEPIDOPTERA, NEPTICULIDAE)

Summary of Doctoral Dissertation Biomedical Sciences, Biology (01B): Entomology and Plant Parasitology, B–250

Vilnius, 2014

1 The work was carried out in 2010–2014 at Lithuanian University of Educational Sciences

Research supervisor: Prof. Habil. Dr. Jonas Rimantas Stonis (Lithuanian University of Educational Sciences, Zoology, 05B and Biology, 01B: Entomology and Plant Parasitology, B–250)

Research advisor: Prof. Dr. Virginijus Sruoga (Lithuanian University of Educational Sciences, Zoology, 05B and Biology, 01B: Entomology and Plant Parasitology, B–250)

Chairman: Prof. Habil. Dr. Rimantas Rakauskas, Vilnius University (Biology, 01B: Entomology and Plant Parasitology, B–250)

Members: Prof. Dr. Sigitas Podėnas, Vilnius University (Biology, 01B: Entomology and Plant Parasitology, B–250)

Dr. David C. Lees, Cambridge University, UK (Biology, 01B: Entomology and Plant Parasitology, B–250)

Dr. M. Alma Solis, the Smithsonian Institution, USA (Biology, 01B: Entomology and Plant Parasitology, B–250)

Assoc. Prof. Dr. Jurga Turčinavičienė, Vilnius University (Biology, 01B: Entomology and Plant Parasitology, B–250)

Assoc. Prof. Dr. Jana Radzijevskaja, Vytautas Magnus University (Biology, 01B)

Dr. Jolanta Rimšaitė, the Nature Research Centre (Biology, 01B: Entomology and Plant Parasitology, B–250)

The defence of the dissertation will be held at a public meeting of the Biology (Entomology and Plant Parasitology) Council at Lithuanian University of Educational

Sciences on September 26, 2014, at 14 p. m. (Auditorium A11). Address: 39 Studentų St., Vilnius, LT–08106, Lithuania.

The summary of the doctoral dissertation was send on August ___, 2014. The dissertation is available at the Library of the Lithuanian University of Educational Sciences.

Agnė Rocienė

KONTINENTINĖS RYTŲ AZIJOS MAŽŲJŲ GAUBTAGALVIŲ (INSECTA, LEPIDOPTERA, NEPTICULIDAE) TAKSONOMIJA

Daktaro disertacijos santrauka Biomedicinos mokslai, Biologija (01B): Entomologija ir augalų parazitologija, B–250

Vilnius, 2014 Disertacija parengta 2010–2014 metais Lietuvos edukologijos universitete.

Mokslinis vadovas: Prof. habil. dr. Jonas Rimantas Stonis (Lietuvos edukologijos universitetas, Zoologija, 05B ir Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Mokslinis konsultantas: Prof. dr. Virginijus Sruoga (Lietuvos edukologijos universitetas, Zoologija, 05B ir Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Pirmininkas: Prof. habil. dr. Rimantas Rakauskas, Vilniaus universitetas (Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Nariai: Prof. dr. Sigitas Podėnas, Vilniaus universitetas (Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Dr. David C. Lees, Kembridžo universitetas, Jungtinė Karalystė (Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Dr. M. Alma Solis, Smitsono centras, JAV (Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Doc. dr. Jurga Turčinavičienė, Vilniaus universitetas (Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Doc. dr. Jana Radzijevskaja, Vytauto Didžiojo universitetas (Biologija, 01B)

Dr. Jolanta Rimšaitė, Gamtos tyrimų centras (Biologija, 01B: entomologija ir augalų parazitologija, B–250)

Disertacija bus ginama viešame Biologijos (entomologijos ir augalų parazitologijos) tarybos posėdyje, kuris vyks 2014 m. rugsėjo 26 d., 14 val., Lietuvos edukologijos universitete, A11 auditorijoje. Adresas: Studentų g. 39, Vilnius, LT–08106, Lietuva

Disertacijos santrauka išsiuntinėta 2014 m. rugpjūčio ___d. Su disertacija galima susipažinti Lietuvos edukologijos universiteto bibliotekoje.

Problem and relevance of the research. Nepticulidae (pygmy moths) are insects whose larvae mine assimilatory tissues of plants, and sometimes they may be pests (or potential pests) of forest and cultivated plants (Diškus & Stonis, 2012). However, this is not the only reason to highlight the importance of pygmy moths. In particular, we should consider the Convention on Biological Diversity, which obliges to inventory the global biodiversity. Currently the biological diversity is rapidly declining because of the climate change and in particular as a result of anthropogenic activities and fragmentation or destruction of habitats suitable for plants and animals (Stonis, 2010, 2011). These processes are not very conspicuous in our country yet, but have gained much greater rate in other regions, including East Asia. The object of the doctoral research is one of the most phylogenetically primitive (and thus theoretically one of the most important) taxa of the Lepidoptera order. The distinctive feature of the Nepticulidae family is deep trophic specialization and wide distribution in almost all terrestrial habitats and continents (except the Antarctic) (Puplesis, 1994; Diškus & Stonis, 2012). Though the family has long been an object of study, the fauna of many regions is still poorly known. East Asia is an important centre of biodiversity and is faunistically related with the European biota. Therefore studies of the East Asiatic Nepticulidae are important for better understanding the old faunogenetic processes which took place in the Eur- asian continent. Furthermore, because of the highly characteristic tendencies of stenophagy, sedentary way of life and endemism, Nepticulidae are among the most suitable tools to characterize the richness of biodiversity, origin and relationships of biomes (or biotas) (Stonis, 2010). However, data on these economically and theoretically important insects are rather poor. Prior to this research, the fauna of Nepticulidae of continental East Asia had not been sufficiently studied (Rocienė & Stonis, 2013; Stonis & Rocienė, 2013). Most of the earlier data had been based on inaccurate descriptions of adult genitalia, and type specimens had been studied and documented relying merely on temporary micro-slides of genitalia (Rocienė & Stonis, 2013). Temporary slides in glycerine, contrary to per-

5 manent slides in Euparal, are less suitable for detailed documentation of overlapping sclerites and comparison of morphological structures. The use of permanent slides in this research not only helped discover new species for science or for the fauna of East Asia but also describe more accurately the distribution ranges of many previously known nep- ticulids and perform the first chorological analysis of the East Asiatic Nepticulidae. Purpose. The purpose of the dissertation is to examine unidentified scientific collection material, carry out fieldwork in continental East Asia, and on the basis of the obtained results, accomplish taxonomic and chorological analysis of the East Asiatic Nepticulidae. Tasks: 1. To identify and name the family’s morphological characters that could and should be used in the diagnostics of the family’s taxa (in particular species) in the context of today’s research on the basis of the new data of Nepticulidae collected in East Asia and new additional (comparative) material collected in the Ukraine, India, Mexico and the countries of Central Asia and tropical Africa. 2. To find out whether the original descriptions of all species are completely precise and characterize the species (to identify and describe particular characters of genitalia if they were found to be inaccurately interpreted or not described) on the basis of the prepared permanent slides in Euparal and the type series of Nepticulidae species described in the fauna of continental East Asia that have been studied and documented. 3. To update morphological characteristics of the previously described species on the basis of the examined unidentified scientific collection material of Nepticulidae collected by other researchers in continental East Asia, performed taxonomic changes and discovered and assessed potential cases of variability of characters. 4. To update the species’ diagnostic characters and data on geographical distribution on the basis of the fieldwork that has been carried out and the examined newly collected material. 5. To discover and describe new species and assess the overall taxonomic diversity of the East Asiatic Nepticulidae on the basis of new material from East Asia.

6 6. To make a chorological analysis of the East Asiatic Nepticulidae and find out how the results of chorological analysis are related with the species’ trophic relationships. 7. By integrating cladistic analysis data and providing additional evidence on the exclusivity of the new group, to designate and describe a new species group of the Stigmella genus on the basis of new material from East Asia. Scientific novelty. A total of 14 Nepticulidae species new to science from East Asia (Russian Far East and NE China), and the Himalaya (In- dia) have been discovered and described or identified and prepared for description. The type series of all 56 Nepticulidae species earlier described in the Russian Far East were examined: permanent morphological micro-slides of the holotypes of all species were made for the first time, and the species descriptions were supplemented with new data and documented according to up-to-date scientific standards. The unidentified Nepticulidae material deposited at the Institute of Zoology of the Russian Academy of Sciences was dissected, examined and described for the first time. The Stigmella regina (Pupl.) was synonymized with S. sashai (Pupl.), and Ectoedemia laura (Pupl.) with E. sivickisi (Pupl.). A taxonomic checklist of the East Asiatic Nepticulidae was updated and supplemented with new species and peculiarities of trophic relationships and geographical distribution of the East Asiatic species were re- viewed. A chorological assessment of the East Asiatic Nepticulidae fauna was done. An annotated taxonomic checklist of the Nepticulidae of Mexico (Central America) was provided for the first time on the basis of the investigation of the fauna of other regions (i.e. comparative studies). The Acalyptris fauna of Central Asia was re-examined in order to disclose morphological characters significant for species taxonomy. A sizeable collection material from tropical Africa was dissected and mounted for the first time. Scientific and practical relevance. In order to understand the extent of the global biodiversity and processes that maintain it, researchers are encouraged to more intensively inventory the biological diversity of the

7 Earth’s biomes and ecosystems and provide taxonomic reviews, revisions, and identification tools (Stonis, 2010, 2011). Our research focus on the leaf-mining (endobiontic) insects is almost equally related to important topics of today’s taxonomy, evolutions and ecology. The family Nep- ticulidae comprises stenophagous (mostly monophagous) species whose larvae, during the whole of the larval stage, feed and develop within green (assimilatory) tissues of plants (Diškus & Stonis, 2012). Such a way of lifestyle is called mining, and the cavities (tunnels) that larvae make in assimilatory tissues are called “leaf-mines” (or simply “mines” if the cavities are in a green tissue of other plant organ than a leaf) (Diškus & Stonis, 2012). Nepticulidae are important from the economical viewpoint as these organisms can be recognized as pest or potential pests (dozens of species have been included into the list of pests of cultivated plants, see Kuznetzov & Puplesis, 1994). Better knowledge of the diversity of pigmy moths fauna is essential for establishing various plant protection meas- ures. On the other hand, Nepticulidae (as some other endobiontic organisms) are rather sedentary and can be detected in nearly all terrestrial ecosystems, though with uneven distribution (Puplesis, 1994). Studying Nepticulidae enables better understanding of evolutionary tendencies and biogeographical peculiarities (Stonis & Rocienė, 2014). The taxonomic inventory of the East Asiatic fauna done in this dissertation presents data which contributes to better understanding of Nepticulidae worldwide, and provides better knowledge of the fauna of East Asia or zoogeographical links (Stonis & Rocienė, 2014; Rocienė & Stonis, 2014), including those with the related Euro-Nemoral fauna. Statements to be defended: 1. The morphological characters of Nepticulidae are important in light of the contemporary research; however, the significance of each character is different; male genital structures are species-specific and stable irrespective of the region where the investigation was carried out. 2. The East Asiatic Nepticulidae are characterized by diverse, though earlier not documented (or erroneously interpreted) characters of genital structures which are important in taxa diagnostics. 3. At least a few of the earlier described East Asiatic species do not exhibit any characters allowing to regard them as independent taxa;

8 therefore, they have to be merged (names synonymized), and morphological characteristics of many other East Asiatic species have to be supplemented. 4. To date, the diagnostics of some East Asiatic species has been based on inaccurate morphological characters, and the geographical distribution of some species has to be supplemented with new essential data. 5. Stigmella multispicata, S. sexcornuta, Ectoedemia ortiva, E. paraortiva, and E. species 219 detected in the Russian Far East, as well as Stig- mella cornuta detected in China, are species new to science. They differ from the earlier known species in clearly expressed characters. A total of 110 Nepticulidae species are currently known in East Asia. Most of them belong to Stigmella and only several to other four genera. 6. The East Asiatic Nepticulidae fauna is not homogeneous chorologically; it is composed of 4 chorological groups with the prevalence of the East-Palaearctic group trophically associated with Quercus, Acer, Malus, and Ulmus; while the Nepticulidae species of broader distribution ranges constitute an insignificant minority. 7. The cornuta group is a new species group of the Stigmella genus. It has some characters common with the S. betulicola group, yet is distinguished by the specific shape and size of cornuti, specific shape of gnathos, partially reduced caudal processes, and the combined mines. Publication of the research results. The results of the research have been published in 12 papers in scientific journals, periodical and one-off publications. Five of them were published in Zootaxa (a journal with an impact factor), one was published in Acta Zoologica Lituanica (a periodical journal included in the Thomson Reuters Master Journal List), one in Ecology and Zoology (ISI, Thomson Reuters Master Journal List), two were papers and abstracts at international scientific conferences, and two appeared in other publications. Materials of the dissertation have been also published in one chapter of a monograph with co-authors (see Diškus & Stonis, 2012: The Nepticulidae (Lepidoptera): Taxonomy, Chor- ological Composition and Trophic Relationships), and in a methodical publication. The author together with co-authors published a total of 10 papers in English and 2 in the Lithuanian language.

9 Structure and scope. The dissertation is written in Lithuanian, with a summary in English. The dissertation is composed of Introduction, Literature Overview, Methods and Materials, Results, Conclusions, and References (328 items). The dissertation consists of 211 pages illustrated with 379 figures. Acknowledgements. My sincere thanks are due to my scientific supervisor Professor Dr. Habil. Jonas Rimantas Stonis (LEU) for a proposal to go deeper into this interesting topic, for support in getting material for research and for versatile remarks and help in preparing this work, and to my scientific advisor Prof. Dr. Virginijus Sruoga (LEU) for valuable advice, encouragement and goodwill. I am grateful to my colleagues from the Department of Biology and in particular of the Biosystem- atics Research Group: Assoc. Prof. Dr. A. Diškus, A. Remeikis and A. Navickaitė, for scientific communication and interesting discussions. I acknowledge the hospitality, friendly cooperation and valuable advice of the Head of the Laboratory DSc. S. Yu. Sinev, Dr. A. L. Lvovsky, Dr. S. V. Baryshnikova and Dr. V. G. Mironov during my visit at the Laboratory of Insect Systematics of the Institute of Zoology of the Russian Academy of Sciences (St. Petersburg). I am also thankful to Dr. J. De Prins, the cura- tor of the Lepidoptera collection at the Royal Museum for Central Africa in Tervuren (Belgium) for making available the material of the Museum. I am sincerely thankful to Prof. Dr. P. S. Zorikov, the Director of the Gor- notayezhnoe Scientific Station of the Far Eastern Division of the Russian Academy of Sciences (Primorskiy Kray), for the possibility to carry out my research and for the technical equipment during the field and laboratory work in 2011. I also thank Dr. M. M. Omelko, the Head of the Laboratory of Insect Ecology of the Gornotayezhnoe Station, and his wife Dr. N. V. Omelko, for hospitality and help. My thanks are due to Prof. Dr. J. Trimble (USA) and Dr. Andrius Petrašiūnas (VU) for their advice, pro- posals to improve the work and sincere discussions. My special thanks are due to my friends and my family, in particular my husband Donatas, my mother Angelė, my father Romas, my godparents Roma and Valdas for endless patience, tolerance, understanding and support. The doctoral studies and fieldwork during the period of my doctoral studies were financed by the EU Structural Funds’ project “Improvement of the Training of High Quality Professionals Who Meet the State and

10 Public Requirements in the Area of Biomedicine (BIOMEDOKT)”, also by the State Studies Foundation and the Research Foundation of the Re- search Council of Lithuania.

LITERATURE OVERVIEW

A detailed overview of literature is presented in 14 printed pages of the dissertation. It contains an overview and analysis of general literature, the main publications of the recent period, and the specific literature on the East Asiatic Nepticulidae.

METHODS AND MATERIALS

Study methods. One of the most effective ways of collecting Neptic- ulidae is light trapping (Diškus & Puplesis, 2003). During our research, adults of pygmy moths were collected at light using a 1.5 × 2.5 m white fabric screen, the upper edge of which was bent by 2 cm and sewn down so that a strong rope could get through the hole. Rope length was about 20–30 m so that the screen could be stretched between trees. A lamp was fastened by a trident holder to the upper edge of the screen (in the centre) above the eye level. DRL or LRF type UV lamps or their analogues were the most suitable for collecting Nepticulidae. The screen was pulled tightly and its lower edge pressed and bent forward. The lower edge on the ground was bent so that fallen moths could be collected. The screen was stretched tightly not to be swung by wind (windy places were avoid- ed). It is easier to collect insects from a tightly stretched fabric than from a badly tensed one where folds make it difficult to notice and collect minute moths. Nepticulidae were taken from the screen by small (about 3.5–5.5 mm in diameter and 3–5 cm in length) glass test-tubes clogged with a cotton wool. Each specimen was put into a separate test-tube so that moths could retain as many scales as possible. The plug of a test-tube had to be tight enough for a moth not to escape and permeable enough to

11 permit the insect-killing chemical (ethyl acetate) to pass through it. The test-tube with a moth was immediately placed into a killing jar – a tightly and easily closed container (usually made of glass) with a 3–5 cm high cotton wool liner on the bottom. In order specimens could be preserved fresh, the collected material was pinned either immediately after a light collecting session or no later than the afternoon of the next day. Before that, in line with the method described in Puplesis, 1994, material was kept in a dark and cool place (e.g. a fridge). Alongside light trapping, adult moths were collected by brushing with a small (15 cm diameter) entomological net over Nepticulidae host- plants. Another important and time and effort consuming way of collecting Nepticulidae material, namely rearing of adults from mining larvae was also used. This method enabled getting not only the highest quality material (i.e. specimens in the best condition, not rubbed) but also important additional data on host-plants, leaf-mines and cocoons (colour and size) as well as the life-cycles (Diškus & Stonis, 2012). In the studies on insects, in particular micro-moths, description of external characters is often insufficient, and slides of morphological structures have to be prepared. Species diagnostics most often relies on the characters of male genitalia (genital apparatus/armature), which, compared with other morphological structures, are distinguished by high specificity (Puplesis & Robinson, 2000). Reliable identification and species description of pygmy moths (in particular new species) is impossible without the examination of genital structures (Diškus & Stonis, 2012). It was noticed that the male genitalia characters of Nepticulidae are little variable and specific at the level of both species or higher taxonomic rank (Šimkevičiūtė [Rocienė] et al., 2010). In line with the methods described by Diškus & Puplesis (2003) and Diškus & Stonis (2012), temporary and permanent micro-slides were prepared. The study of genitalia mounts (micro-slides) was performed by using high magnification microscopes. Genital structures were measured and photographed, and sclerites were described. All micro-slides were recorded in a special register, providing the slide number, name of the species, dissection date, institution to which the dissected specimen belongs, and other information.

12 The lengths of genital structures important for diagnostics were measured both in temporary (in glycerine) and permanent (in Euparal) mounts by a Leica DM 2500 stereoscopic microscope, Leica DFC 420 digital camera, and the LeicaApplicationSuite V3.0.0 software. Adults were measured by a MBS-10 stereoscopic microscope with an ocular measuring ruler, and their exterior was photographed by a Leica S6D stereoscopic microscope, DFC 290 camera, and the LeicaImageManager IM50 software. The distribution ranges of species were thoroughly mapped and chorologically analysed. Taxa distinguished for the same or similar distribution range were joined into groups which, in their turn, were named indicating the type of the range. For cladistics analysis as many characters as possible were used including not only the morphological characters but also chorological and ecological ones. Though different numbers of synapomorphic and autapomorphic characters were chosen for different clades the higher number of synapomorphic characters is more reliably indicative the fact of common ancestor. Materials. Scientific material for the dissertation was collected by the author in 2010–2013 (Figs 1, 2) during the fieldwork in the Himalaya (2010), Russian Far East (mainly at Gornotayezhnaya Station of the Far East Division of the Russian Academy of Sciences (Ussuriysk District, 2011), and northeastern China (2013); an additional “search-trip” to northern Thailand was organized in 2012. The types of 56 East Asiatic species and a sizeable unidentified Nep- ticulidae material for our scientific studies was temporary borrowed from the Institute of Zoology of the Russian Academy of Sciences and the Karadag Biological Station (Fig 3). In addition, the large comparative material collected by other researchers in Mexico (Šimkevičiūtė [Rocienė] et al., 2009), Ukraine and countries of Central Asia and tropical Africa (Figs 2–5) is examined and used in the morphological chapter of the dissertation. A total of about 3500 Nepticulidae specimens were examined. Hun- dreds of temporary and over 600 permanent mounts (micro-slides) of genitalia were prepared.

13 Fig. 1. Map of the region (courtesy of T. Patterson, USA) showing the collection localities by the dissertation author. In addition, a sizeable material was collected by various other researchers in various other sites (not shown on the map): Andreevka, 42°38′ N, 131°07′ E; Barabash Levada, 44°45′ N, 131°25′ E; Kedrovaya Pad‘ Nature Reserve, 43°11′ N, 131°24′ E; Kamenushka, 44°10′ N, 131°22′ E; Ryazanovka, 42°47′ N, 131°14′ E; Zanadvorovka, 43°18′ N, 131°37′ E, and a few other localities.

14 Figs 2–5. Maps of the regions (courtesy of T. Patterson, USA) showing the collection localities of the additional, comparative material: 2 – in Central Asia and the Himalaya; 3 – in the Crimea; 4 – in tropical Africa; 5 – in Central America (Mexico).

15 RESULTS

Investigation of morphological structures of Nepticulidae. In some cases, new investigation methods are given too much attention and it is forgotten that taxonomic work is impossible without traditional analysis of morphology and biological peculiarities of organisms (Beutel & Kristensen, 2012). The current system of Nepticulidae and species diagnostics has been so far mostly based on morphological and biological data. Scientific literature on morphological characters of Nepticulidae is rather numerous (Scoble, 1983; van Nieukerken, 1986a; Johansson et al., 1990; Puplesis, 1994a; Puplesis & Robinson, 2000; Puplesis & Diškus, 2003, etc.). An original overview of morphological characters was prepared. The morphological analysis of the main Nepticulidae material collected in East Asia and additional (comparative) material collected in the Ukraine, India, Mexico, and the countries of Central Asia and tropical Africa showed that the shape of valva, transtilla, vinculum, and uncus lobes, the number and shape of gnathos processes are also essential diagnostic characters of the Nepticulidae species. The stability of these characters was registered after the analysis and comparison of material from diverse and remote regions of the Earth; intra-specific or other variability of the above-mentioned structures that might affect the reliable species diagnostics was not found. The results of these investigations were presented together with co-authors in Stonis et al. (2012) and in a monograph (Diškus & Stonis, 2012). For a detailed overview of the morphology of Nepticulidae, see pp. 35–41 of the dissertation. Results of the study of the type series of the Nepticulidae species described from continental East Asia. The current diagnostics of the East Asiatic Nepticulidae species is mostly based on male genital structures, which are little variable and are species-specific; therefore, a detailed study and precise representation of such structures is very important for reliable species diagnostics. Before our dissertation studies, East Asia was known to comprise 103 Nepticulidae species. Most of them (63 species) were described from continental East Asia, i.e. Primorskiy Kray, Russian Far East. All these Nepticulidae species were described exclusively on the basis of temporary mounts (slides) of genital structures in glycerine. Most of such slides not always precisely demonstrated the main diagnostic characters of the species, and the element of sub- jectivity often occurred (also because of the poor quality of microscopes). The

16 drawings of genital structures were not sufficiently precise and sometimes even confusing. For the first time and by using up-to-date entomological methods, we dissected, re-examined, and documented the type material of the earlier described Nepticulidae species from East Asia, mainly from Primorskiy Kray (also from a few adjacent territories) and provided the first photographic documentation of the types of 56 species described by R. Puplesis (Stonis & Rocienė, 2013). The morphology of most re-examined species was considerably updated (the classification presented below follows Puplesis, 1994; Puplesis & Robinson, 2000; Puplesis et al., 2002; and Puplesis & Diškus, 2003). 1. Stigmella dissona (Puplesis, 1984) (Astigmella dissona Puplesis, 1984a: 112), a junior syn. of Stigmella naturnella (Klimesch, 1936). Mor- phology update (Figs 1–4 in Stonis & Rocienė, 2013): Gnathos deeply divided. System of cornuti complex (Figs 1, 3 in Stonis & Rocienė, 2013). 2. Stigmella mirabella (Puplesis, 1984) (Astigmella mirabella Puplesis, 1984a: 112). Morphology update (Figs 5–8 in Stonis & Rocienė, 2013): Valva without pointed apex. Gnathos divided. Vesica with 8–9 large spines (Figs 7, 8 in Stonis & Rocienė, 2013). 3. Stigmella kurilensis Puplesis, 1987: 8. Recently was studied and photographed by van Nieukerken & Kuroko 2005: 31–35, Figs 5–10. 4. Stigmella sashai Puplesis, 1984b: 594–596. Morphology update (Figs 9–12 in Stonis & Rocienė, 2013): Valva with developed inner lobe. Transverse bar of transtilla interrupted. Cornuti spine-like; the vesica partially everted in the holotype (Fig. 12 in Stonis & Rocienė, 2013). 5. Stigmella regina Puplesis, 1984b: 596. Morphology update (Figs 13–16 in Stonis & Rocienė, 2013): Phallus with two clusters of spine-like cornuti (Fig. 15 in Stonis & Rocienė, 2013). 6. Stigmella sakhalinella Puplesis, 1984a: 115, 116. Morphology update (Figs 17, 18 in Stonis & Rocienė, 2013): Valva pointed apically. Uncus with lateral lobes. Phallus with apical cornuti (Fig. 17 in Stonis & Rocienė, 2013). 7. Stigmella attenuata Puplesis 1985: 62. Morphology update (Figs 20, 21 in Stonis & Rocienė, 2013): Phallus with apical cornuti. Uncus bilobed. 8. Stigmella ultima Puplesis, 1984a: 117. Morphology update (Figs 22, 23 in Stonis & Rocienė, 2013): Valva with papillae on inner lobe. Cornuti small, mostly denticulate. 9. Stigmella tegmentosella Puplesis, 1984a: 117. Morphology update (Figs 24–26 in Stonis & Rocienė, 2013): Valva with papillae and spine-like

17 process on inner lobe. Cornuti small, mostly denticulate (Fig. 24 in Stonis & Rocienė, 2013). 10. Stigmella monella Puplesis, 1984a: 117, 118. Morphology update (Figs 27–31 in Stonis & Rocienė, 2013): Valva with undeveloped inner lobe and spine-like process. Transtilla interrupted medially. Cornuti minute, mostly denticulate (Fig. 31 in Stonis & Rocienė, 2013). 11. Stigmella kozlovi Puplesis, 1984a: 118, 119. Morphology update (Figs 32–35 in Stonis & Rocienė, 2013): Valva with papillae on inner lobe. Transtilla interrupted medially. Cornuti minute, mostly denticulate (Fig. 35 in Stonis & Rocienė, 2013). 12. Stigmella nostrata Puplesis, 1984a: 113, 114. Morphology update (Figs 36–40 in Stonis & Rocienė, 2013): Valva with large inner lobe. Gna- thos with relatively broad caudal processes. 13. Stigmella aurora Puplesis, 1984a: 119. Morphology update (Figs 41–44 in Stonis & Rocienė, 2013): Valva with triangular process on inner lobe. Gnathos with large anterior processes. Cornuti small, denticulate, spine-like to irregular (Fig. 44 in Stonis & Rocienė, 2013). 14. Stigmella micromelis Puplesis, 1985: 59, 60. Morphology update (Figs 45–49 in Stonis & Rocienė, 2013): A more thorough examination of the male genitalia of the holotype reveals very little difference from that of the original description (Puplesis 1985: fig. 4 and Puplesis 1994: fig. 300). 15. Stigmella crataegivora Puplesis, 1985: 60–62. Morphology update (Figs 50–53 in Stonis & Rocienė, 2013): A more thorough examination of the male genitalia of the holotype shows little difference from that of the original description (Puplesis 1985: fig. 3 and Puplesis 1994: fig. 297). Gnathos with larger caudal and anterior processes (Fig. 52 in Stonis & Rocienė, 2013). 16. Stigmella taigae Puplesis, 1984a: 112, 113. Morphology update (Figs 54–56 in Stonis & Rocienė, 2013): Valva with papillae on inner lobe. Phallus with larger clusters of spine-like cornuti. 17. Stigmella palionisi Puplesis, 1984b: 596. Morphology update (Figs 65–67 in Stonis & Rocienė, 2013): Juxta developed, with narrow sclerotized apex. Phallus with two lateral groups of larger, sclerotized cornuti and numerous tiny denticulate cornuti in between. 18. Stigmella amuriella Puplesis, 1985: 62. Morphology update (Figs 57–59 in Stonis & Rocienė, 2013): Valva with rounded inner lobe. Phallus with slender spine-like cornuti.

18 19. Stigmella alisa Puplesis 1985: 63. Morphology update (Figs 60–64 in Stonis & Rocienė, 2013): Juxta with narrow apex. Gnathos with long anterior processes (Figs 63, 64). Phallus with about 10 large cornuti and numerous small spine-like cornuti arranged in two lateral groups. 20. Stigmella auricularia Puplesis, Diškus & Juchnevič, 2003: 245, 246. Morphology update (Figs 68–75 in Stonis & Rocienė, 2013): Uncus with triangular lateral lobes. Ventral plate of vinculum with deep oval excavation (Figs 68, 71, 74, 75 in Stonis & Rocienė, 2013). 21. Stigmella ussurica Puplesis, 1987: 8–10; a junior syn. of Stigmella tranocrossa Kemperman & Wilkinson, 1985. Morphology update (Figs 76–79 in Stonis & Rocienė, 2013): Uncus with two large median and two smaller lateral lobes. Phallus with three large apical cornuti and a compact group of 5 smaller basal cornuti (Figs 76, 79 in Stonis & Rocienė, 2013). 22. Stigmella uigurica Puplesis, 1985: 62, 63; a junior syn. of Stigmella continuella (Stainton, 1856). Morphology update (Figs 80–84 in Stonis & Rocienė, 2013): Lateral lobes of vinculum very long and narrow. Valva with two short apical processes. Phallus with numerous tiny denticulate cornuti and a few spine-like cornuti (Figs 82–84 in Stonis & Rocienė, 2013). 23. Stigmella magica Puplesis, 1985: 63–65; a junior syn. of Stigmella lediella (Schleich, 1867). Morphology update (Figs 85–88 in Stonis & Ro- cienė, 2013): Inner lobe of valvae with numerous papillae. Apical process of valva very long and curved. Phallus with about 7 apical and 7 basal cornuti (Fig. 86 in Stonis & Rocienė, 2013). 24. Stigmella rhododendri Puplesis, 1985: 65; a junior syn. of Stig- mella lediella (Schleich, 1867). This species was described from another host-plant (Rhododendron schlippenbachii Maxim.); male genialia with broad sublateral processes of transtilla and more numerous basal cornuti (Puplesis 1985). However, the holotype was not available for our study. 25. Stigmella palmatae Puplesis, 1984a: 115. Morphology update (Figs 89, 90 in Stonis & Rocienė, 2013): Outer margin of valva sinuous, inner lobe papillated, apical process blunt. Phallus missing (Stonis & Ro- cienė, 2013). 26. Stigmella monticulella Puplesis, 1984a: 114. Morphology update (Figs 91–93 in Stonis & Rocienė, 2013): Valva with slender apical process. Phallus with manica.

19 27. Stigmella dentatae Puplesis, 1984a: 114, 115. Morphology update (Figs 94, 95 in Stonis & Rocienė, 2013): Apical cornuti large, spine-like. Uncus may be concealed in slide-mounted preparations (Stonis & Rocienė, 2013). 28. Stigmella aladina Puplesis, 1984a: 115. Morphology update (Figs 98– 101 in Stonis & Rocienė, 2013): Valva with rounded inner lobe. Phallus with a large apical group of spine-like cornuti (Fig. 99 in Stonis & Rocienė, 2013). 29. Stigmella omelkoi Puplesis, 1984b: 593. Morphology update (Figs 96, 97 in Stonis & Rocienė, 2013): Uncus appearing short if bent downward in preparations. Uncus with broad lateral processes (Fig. 97 in Stonis & Rocienė, 2013). Valva with rounded inner lobe. Phallus with very large manica (Fig. 96 in Stonis & Rocienė, 2013). 30. Stigmella fervida Puplesis, 1984b: 593, 594. Morphology update (Figs 102–105 in Stonis & Rocienė, 2013): Lateral processes of gnathos broad basally. Ventral plate of vinculum broad medially, shorter laterally. Phallus with large, lobe-like apical cornutus and numerous spine-like cornuti (Figs 103, 105 in Stonis & Rocienė, 2013). 31. Stigmella kurokoi Puplesis, 1984b: 594. Morphology update (Figs 106–109 in Stonis & Rocienė, 2013): Inner lobe of valvae heavily papillated, apical process pointed. Phallus with manica. 32. Bohemannia manschurella Puplesis, 1984b: 587. Morphology update (Figs 110–113 in Stonis & Rocienė, 2013): Process of juxta bifurcate distally. Phal- lus with one large, spine-like, apical cornutus (Fig. 113 in Stonis & Rocienė, 2013). 33. Bohemannia ussuriella Puplesis, 1984b: 588. Morphology update (Figs 114–117 in Stonis & Rocienė, 2013): Juxta with pointed lateral lobes. Phallus with large spine-like apical cornuti. 34. Bohemannia suiphunella Puplesis, 1984b: 588. Morphology update (Figs 118–121 in Stonis & Rocienė, 2013): Ventral plate of vinculum very short and rounded. Phallus with apical group of spine-like processes (Figs 120, 121 in Stonis & Rocienė, 2013). 35. Bohemannia nubila Puplesis, 1985: 69–71. Morphology update (Figs 122–127 in Stonis & Rocienė, 2013): Valva almost triangular, with pointed apical process and papillated inner lobe. Ventral plate of vinculum narrowed distally (Figs 122, 125 in Stonis & Rocienė, 2013). 36. Bohemannia piotra Puplesis, 1984b: 586, 587. Morphology update (Figs 128–131 in Stonis & Rocienė, 2013): Valva broad, with very slender apical process. Phallus with spine-like cornuti.

20 37. Ectoedemia emendata Puplesis, 1985: 69; a junior syn. of Ectoe- demia amani Svensson, 1966: 200, 201, fig. 34, pl. 4, fig. 3. The holotype of E. emendata was not available for our study. 38. Ectoedemia admiranda Puplesis, 1984b: 588, 590. Morphology update (Figs 132–139 in Stonis & Rocienė, 2013): Valva abruptly narrowed in apical part. Phallus with three pairs of carinae, numerous tiny cornuti, and long band of spine-like cornuti (Fig. 138 in Stonis & Rocienė, 2013). 39. Ectoedemia sivickisi Puplesis, 1984b: 590. Morphology update (Figs 140–143 in Stonis & Rocienė, 2013): Sublateral processes of transtilla very long. Phallus with three pairs of carinae and numerous tiny cornuti (Figs 140, 142 in Stonis & Rocienė, 2013). 40. Ectoedemia laura Puplesis, 1985: 68, 69. Morphology update (Figs 144–148 in Stonis & Rocienė, 2013): Sublateral processes of transtilla very long. Phallus with three pairs of carinae and numerous tiny cornuti on vesica (everted in the holotype) (Figs 145, 147 in Stonis & Rocienė, 2013). 41. Ectoedemia arisi Puplesis, 1984a: 120–122. Morphology update (Figs 149–152 in Stonis & Rocienė, 2013): Valva papillated. Phallus with single, very large dorsal carina. 42. Ectoedemia christopheri Puplesis, 1985: 69 (originally described as Ectoedemia wilkinsoni Puplesis, 1984a: 122), a junior homonym of Ectoe- demia wilkinsoni Scoble, 1983. Morphology update (Figs 153–160 in Stonis & Rocienė, 2013): Caudal process of gnathos strongly sclerotized and appearing truncate at apex. Valvae with numerous papillae and long setae (Fig. 155 in Stonis & Rocienė, 2013). Transverse bar of transtilla thickened. Phallus with narrow band of spines between lateral carinae; two pairs of carinae (lateral and ventral) pointed distally (Figs 157–160 in Stonis & Rocienė, 2013), except single sublateral lobe (sublateral carina) (Fig. 160 in Stonis & Rocienė, 2013). 43. Ectoedemia philipi Puplesis, 1984b: 590, 592. Morphology update (Figs 161, 162 in Stonis & Rocienė, 2013): Gnathos with elaborate and strongly slerotized lateral arms. Phallus rounded apically, with two pairs of spine-like carinae (Figs 161, 162 in Stonis & Rocienė, 2013). 44. Ectoedemia chasanella Puplesis, 1984a: 124. Morphology update (Figs 163–168 in Stonis & Rocienė, 2013): Caudal process of gnathos nar- rowly to broadly rounded distally. Phallus with two triangular ventral carinae and some minute cornuti on vesica (everted in Figs 167, 168 in Stonis & Rocienė, 2013).

21 45. Ectoedemia scoblei Puplesis, 1984a: 122. Morphology update (Figs 169–172 in Stonis & Rocienė, 2013): Valva heavily papillated on inner margin. Lateral arms of gnathos very long and broadened distally. Phal- lus with two ventral carinae (Figs 171, 172 in Stonis & Rocienė, 2013). 46. Ectoedemia aligera Puplesis, 1985: 67, 68. Morphology update (Figs 173–178 in Stonis & Rocienė, 2013): Inner margin of valva papillated. Lateral arms of gnathos long and broadened distally. Ventral carinae of phallus comprise two (Fig. 175 in Stonis & Rocienė, 2013) to five slender spine-like processes (Fig. 177 in Stonis & Rocienė, 2013). 47. Ectoedemia ermolaevi Puplesis, 1985: 68. Morphology update (Figs 179, 180 in Stonis & Rocienė, 2013): Valva with strongly slerotized, pointed apical process. Phallus with two ventral carinae (Fig. 180 in Stonis & Rocienė, 2013). 48. Ectoedemia maculata Puplesis, 1987: 11, 12. Morphology update (Figs 181, 182 in Stonis & Rocienė, 2013): Apical process of valva triangular. Pseuduncus very large, broadly rounded (partially bent in Fig. 182 in Stonis & Rocienė, 2013). 49. Ectoedemia olvina Puplesis, 1984a: 119, 120. Morphology update (Figs 183–185 in Stonis & Rocienė, 2013): Valva papillated on inner margin. Phallus with two ventral carinae. 50. Ectoedemia ornatella Puplesis, 1984a: 120. Morphology update (Figs 186–190 in Stonis & Rocienė, 2013): Valva papillated on inner margin and apical process. Phallus with two ventral carinae (Figs 188, 189 in Stonis & Rocienė, 2013) and some minute cornuti on vesica (Fig. 190 in Stonis & Rocienė, 2013). 51. Ectoedemia ivinskisi Puplesis, 1984a: 120. Morphology update (Figs 191, 192 in Stonis & Rocienė, 2013): Apical process of valva heavily papillated. Gnathos without median excavation or processes (Figs 191, 192 in Stonis & Rocienė, 2013). 52. Ectoedemia pilosae Puplesis, 1984a: 123, 124. The holotype was not available for our study. 53. Ectoedemia picturata Puplesis, 1985: 65, 67. Morphology update (Figs 193, 194 in Stonis & Rocienė, 2013): Pseuduncus very long and broadly rounded distally. Valva broad, with very slender apical process. Phallus with two divided ventral carinae and some minute cornuti on vesica. 54. Ectoedemia sinevi Puplesis, 1985: 67. Morphology update (Figs 195–199 in Stonis & Rocienė, 2013): Valva gradually narrowed towards

22 apex, with pointed apical process. Phallus with seven slender spine-like carinae (Figs 196, 197, 199 in Stonis & Rocienė, 2013). 55. Ectoedemia insularis Puplesis, 1985: 68. Morphology update (Figs 200–210 in Stonis & Rocienė, 2013): Valva with triangular apical process, heavily papillated. Gnathos with large caudal process and broad lateral arms. Phallus with two slender ventral and two lateral carinae; minute cornuti on vesica (Fig. 210 in Stonis & Rocienė, 2013). 56. Fomoria permira Puplesis, 1984b: 592, 593. Morphology update (Figs 211–222 in Stonis & Rocienė, 2013): Phallus with four very long and slender cornuti, two very long carinae, and two short, distally pointed carinae. Cathrema well developed, its pointed sublateral corners can be misstaken for cornuti (Figs 214, 218, 220 in Stonis & Rocienė, 2013). 57. Etainia capesella (Puplesis, 1985) (Obrussa capesella Puplesis, 1985 in Puplesis & Ivinskis 1985: 39, 40). Morphology update (Figs 223–226 in Stonis & Rocienė, 2013): Inner margin of valva sinuous. Gnathos with two short triangular caudal processes. Phallus with two very large apical cornuti and two apical lobe-like carinae; cathrema well developed, complex, and very large. 58. Etainia tigrinella (Puplesis, 1985) (Obrussa tigrinella Puplesis, 1985 in Puplesis & Ivinskis 1985: 40, 41). Morphology update (Figs 227, 228 in Stonis & Rocienė, 2013): Pseuduncus distinctly truncate. Phallus with one very large, basally lobed, distally pointed cornutus and four apical carinae; cathrema well developed, complex, and very large (Fig. 224 in Stonis & Rocienė, 2013). 59. Etainia peterseni (Puplesis, 1985) (Obrussa peterseni Puplesis, 1985 in Puplesis & Ivinskis 1985: 41–43). Morphology update (Figs 231– 235 in Stonis & Rocienė, 2013): Valva with slender pointed inner process. Phallus with three very large and one small cornuti, and two strongly sclerotized lateral and one lobe-like, median carinae; cathrema well developed, complex, and very large (Fig. 230 in Stonis & Rocienė, 2013). 60. Etainia sabina (Puplesis, 1985) (Obrussa sabina Puplesis, 1985 in Puplesis & Ivinskis 1985: 43, 44). Morphology update (Figs 229, 230 in Stonis & Rocienė, 2013): Valva without apical process (Fig. 229). Phallus with two very large and two smaller apical cornuti and two lateral, distally sclerotized and pointed carinae; cathrema well developed, complex, and very large. If compared with the original descriptions (given in Puplesis 1984a, 1984b, 1985, 1987, Puplesis & Ivinskis 1985; Puplesis & Diškus 2003),

23 only the Etainia, a few of Stigmella (particularly S. micromelis, S. crate- givora) and a few Ectoedemia species exhibit very little difference in morphology of the genitalia. The present study is rather significant because these tiny pygmy moths have not been extensively studied and, therefore, many species still remain to be discovered and described. Without the baseline data providing morphology of genital armature of the type material, it was impossible to continue studies on the East Asiatic fauna of the Nepticulidae. Results of the study of the material deposited at the Institute of Zoology of the Russian Academy of Sciences. Before the dissertation studies, in 1974–1990, sizeable material was collected in the Russian Far East (mainly Primorskiy Kray, also from adjacent territories) by S. Yu. Sinev, S. V. Seksyaeva (Baryshnikova), R. Puplesis, M. M. Omelko and others in: Andreevka, 42°38′ N, 131°07′ E, Barabash Levada, 44°45′ N, 131°25′ E, Kedrovaya Pad‘ Nature Reserve, 43°11′ N, 131°24′ E, Kamen- ushka, 44°10′ N, 131°22′ E, Ryazanovka, 42°47′ N, 131°14′ E, Zanadvo- rovka, 43°18′ N, 131°37′ E, and a few other localities (Rocienė & Stonis, 2013). The material was deposited at the Institute of Zoology of the Russian Academy of Sciences and was transferred to us for the purpose of this dissertation. The author’s earlier analysis of the type series of the previously described species and the new assessment of morphological characters by the author provided grounds for the identification and taxonomic analysis of this unstudied material. A total of 1000 specimens were dissected, analysed and identified. The result of the taxonomic analysis has yielded 35 species (including two species new to science: Ectoe- demia ortiva Rocienė & Stonis and E. species 219). Stigmella naturnella (Klimesch, 1936). Male genitalia of the holotype of Astigmella dissona Puplesis, 1984 (a junior synonym of S. naturnella) were re-examined in the preceding paper (Stonis & Rocienė 2013: figs 1, 3). However, the additional material of S. naturnella collected at the same locality as the holotype of Astigmella dissona shows some variation in the genitalia: rounded capsule, strong lateral scleotization of uncus, more complex but compact cornuti (Figs 5–8 in Rocienė & Stonis, 2013). Stigmella sashai Puplesis, 1984 (Stigmella regina Puplesis, 1984b: 596, new synonymy). From the study of the holotypes of Stigmella regina and S. sashai and three non-type specimens listed in Rocienė & Stonis, 2013 (Figs

24 9, 10, 12, 13), it is clear that they are conspecific. Cornuti form three groups: 1) a dense cluster (sometimes divided) of strongly sclerotized spine-like cornuti, 2) loose, weakly slerotized spine-like cornuti, and 3) minute denticulate cornuti. The vesica in the holotype ofS. sashai is partially everted; therefore it looks somewhat misleading (Fig. 11 in Rocienė & Stonis, 2013). Other characters mentioned by Puplesis (1984b) are not consistent when more material is examined: shape of valvae and uncus are variable; moreover, artefacts of slide-mounting affect the appearance of these structures. Therefore we synonymize Stigmella regina (Rocienė & Stonis, 2013). Stigmella betulicola (Stainton, 1856). In East Asia this species formerly was recorded only from Japan (Kemperman & Wilkinson, 1985). How- ever, a single specimen collected in Kamenushka (Ussuriysk District, Primorskiy Kray) confirms its occurrence on continental East Asia (Figs 14–17 in Rocienė & Stonis, 2013) (new record in Rocienė & Stonis, 2013). Stigmella sakhalinella Puplesis, 1984. The apical cornuti of the phallus of S. sakhalinella are partially broken in the neotype (see Stonis & Ro- cienė 2013: figs 17, 19). Therefore, we confirm morphological details of the male genitalia based on non-type material collected at the same locality as the neotype (Figs 18, 19 in Rocienė & Stonis, 2013). Stigmella attenuata Puplesis 1985. The valvae are partially broken in the holotype of S. attenuata (see Stonis & Rocienė 2013: figs 20, 21). Therefore, we confirm morphological details of the male genitalia based on non-type material collected at the same locality as the holotype (Figs 20, 21 in Rocienė & Stonis, 2013). Stigmella cathepostis Kemperman & Wilkinson, 1985. This species was previously known only from Kyushu, Japan (Kemperman & Wilkin- son 1985). However, two specimens collected in Gornotayezhnoe (20 km E Ussuriysk) and Ryazanovka (Khasan District) confirm the occurrence (and possibly a wide distribution) of S. cathepostis in Primorskiy Kray and continental East Asia (Figs 22–25 in Rocienė & Stonis, 2013) (new record in Rocienė & Stonis, 2013). Stigmella ultima Puplesis, 1984 (see Rocienė & Stonis, 2013). Stigmella kozlovi Puplesis, 1984 (see Rocienė & Stonis, 2013). Stigmella palionisi Puplesis, 1984 (see Rocienė & Stonis, 2013). Stigmella alisa Puplesis 1985. The genital capsule of S. alisa is disarticulated in the holotype (see Stonis & Rocienė 2013: figs 60–64). There-

25 fore, we provide additional photographic documentation of the male genitalia of S. alisa based on non-type material but collected at the same locality as the holotype (Figs 32–36 in Rocienė & Stonis, 2013). Stigmella auricularia Puplesis, Diškus & Juchnevič, 2003. Our previ- ous study of the holotypes of S. auricularia and S. palionisi (see Stonis & Rocienė 2013: figs 65–75) and the present study of seven non-type specimens of S. auricularia and S. palionisi collected in Gornotayezhnoe (20 km E Ussuriysk) confirm the concept of two separate species. In addition to the characters mentioned by Puplesis (1984b) and Puplesis & Diškus (2003), the vinculum has large triangular lobes in S. auricularia (see Rocienė & Stonis, 2013: figs 26–28) but very tiny ones in S. palionisi (see Rocienė & Stonis, 2013: figs 29–31). Stigmella dentatae Puplesis, 1984. The genital capsule of S. dentatae is slightly flattened in the holotype (see Stonis & Rocienė 2013: figs 94, 95). Therefore, we provide additional photographic documentation of the male genitalia of S. dentatae based on non-type material but collected at the same locality as the holotype (see Rocienė & Stonis, 2013: figs 37, 38). Stigmella aladina Puplesis, 1984. The genital capsule of S. aladina is flattened in the holotype (see Stonis & Rocienė 2013: figs 98–101). There- fore, we provide additional photographic documentation of the male genitalia of S. aladina based on non-type material (see Rocienė & Stonis, 2013: figs 39, 40). Stigmella omelkoi Puplesis, 1984 (see Rocienė & Stonis, 2013). Stigmella fervida Puplesis, 1984 (see Rocienė & Stonis, 2013). Bohemannia manschurella Puplesis, 1984 (see Rocienė & Stonis, 2013). Bohemannia ussuriella Puplesis, 1984 (see Rocienė & Stonis, 2013). Bohemannia nubila Puplesis, 1985(see Rocienė & Stonis, 2013). Ectoedemia amani Svensson, 1966 (see Rocienė & Stonis, 2013). Ectoedemia admiranda Puplesis, 1984 (see Rocienė & Stonis, 2013). Ectoedemia sivickisi Puplesis, 1984 (Ectoedemia laura Puplesis, 1985: 68, 69, new synonymy in Rocienė & Stonis, 2013). Based on the examination of the holotypes of Ectoedemia laura and E. sivickisi (Stonis & Ro- cienė 2013) and six non-type specimens listed in Rocienė & Stonis, 2013: figs 41–46, it is clear that they are conspecific. The characters E.of laura mentioned by Puplesis (1985) are not consistent when more material is examined: shape of valvae and uncus varies slightly; moreover, artefacts

26 of slide-mounting influence the appearance of these structures. Therefore we synonymize E. laura (Rocienė & Stonis, 2013). Ectoedemia arisi Puplesis, 1984 (see Rocienė & Stonis, 2013). Ectoedemia christopheri Puplesis, 1984 (see Rocienė & Stonis, 2013). Ectoedemia chasanella Puplesis, 1984. We provide the first photographic documentation of the male genitalia of E. chasanella based on non-type material (see Rocienė & Stonis, 2013: figs 47, 48). Ectoedemia scoblei Puplesis, 1984. We provide the first photographic documentation of the male genitalia of E. scoblei based on non-type material (see Rocienė & Stonis, 2013: fig. 61). Difference between the non- type specimen (slide no. AG 355, Fig. 61 by Rocienė & Stonis, 2013) and the holotype (see Stonis & Rocienė 2013: figs 169–172) are likely the result of slide-mounting of the genitalia. Ectoedemia olvina Puplesis, 1984. We provide the first photographic documentation of the male genitalia of E. olvina based on non-type material (Rocienė & Stonis, 2013: figs 49, 50). Although similar to E. olvina, the male genitalia of the specimen 224 (Rocienė & Stonis, 2013: figs 52– 55) differ from other non-type specimens ofE. olvina (Rocienė & Stonis, 2013: figs. 49, 50) and the holotype, particularly in the short, strongly papillated apical process of the valva (see Rocienė & Stonis, 2013: fig. 52). Ectoedemia ornatella Puplesis, 1984. The genital capsule of the holotype of E. ornatella is partially disarticulated, with the gnathos turned anteriorly (see Stonis & Rocienė 2013: figs 186, 187). Therefore, we provide additional photographic documentation of the male genitalia of the species based on non-type material from the new locality: Andreevka, Khasan District (Rocienė & Stonis, 2013: figs 56–59) (new record in Ro- cienė & Stonis, 2013). Ectoedemia ivinskisi Puplesis, 1984a (see Rocienė & Stonis, 2013). Ectoedemia picturata Puplesis, 1985. The genital capsule of the holotype of E. picturata is disarticulated and partially broken (see Stonis & Rocienė 2013: figs 193, 194). Therefore, we provide additional photographic documentation of male genitalia of the species based on non-type material (Rocienė & Stonis, 2013: fig. 60) collected in the same locality as the holotype. Etainia capesella (Puplesis, 1985). The uncus and gnathos are barely visible in the holotype of E. capesella (see Stonis & Rocienė 2013: figs 223,

27 224). Therefore, we provide additional photographic documentation of the male genitalia of E. capesella based on a non-type specimen (Rocienė & Stonis, 2013: figs 62, 63) collected at a slightly different locality from the holotype: Kamenushka, Ussuriysk District. Etainia tigrinella (Puplesis, 1985) (see Rocienė & Stonis, 2013). Etainia peterseni (Puplesis, 1985) (see Rocienė & Stonis, 2013). Etainia sabina (Puplesis, 1985) (see Rocienė & Stonis, 2013). The examination of the material showed that two species which earlier had been regarded as separate (Stigmella regina, Ectoedemia laura) must not be longer recognized as independent taxa; therefore, S. sashai was synonymized with S. regina, and Ectoedemia sivickisi with E. laura (Rocienė & Stonis, 2013). The synonymity of these names was supported by other researchers as well (van Nieukerken, pers. comm.). The data on distribution of several species detected in the material were updated, and new data on distribution of three species (Ectoedemia ornatella, Etainia capesella, Stigmella betulicola) were disclosed. In addition, a few species that were new to science were discovered and described; they are presented in a separate section of the dissertation. A detailed morphological analysis of all examined species was performed, and new (not described earlier) morphological characters significant for taxonomy and diagnostics were determined (see Rocienė & Stonis, 2013). This taxonomic re-examination of sizeable material provides the foundation for updating the taxonomic checklist of the East Asiatic Nep- ticulidae. Results of the fieldwork in continental East Asia. Before our fieldwork, the Nepticulidae fauna East Asia comprised 105 species, including 67 species occuring in the Russian Far East and 53 in Japan (20 of which occur both in Japan and Russia). Very little is known about the Neptic- ulidae of Korea and China (see Park, 1983; van Nieukerken & Liu, 2000; Puplesis & Diškus, 2003; Lee & Byun, 2007). The fieldwork in Primorskiy Kray as well as the dissection and examination of the collected material provided new data on Nepticulidae of this region, their distribution and morphological characters. In addition, three species new to science were identified – two of the Stigmella Schrank genus and one Ectoedemia Bus- ck (they are presented in the dissertation’s section on species that are new to science).

28 Stigmella thuringiaca (Petry) was identified for the fauna of East Asia for the first time. Earlier it was known only from Europe. Male genitalia of the East Asiatic specimen does not show any significant differences from European examples. Vinculum with triangular lateral lobes. Uncus deeply divided into two large lobes. Gnathos with broad angular plate and two caudal processes. Valva with bulged inner lobe and pointed, inwardly bent apical process; transtilla with slender short sublateral processes. Phallus broad, vesica with very large cluster of slender spine-like cornuti (see the dissertation or the authors’ co-authored paper (Stonis & Rocienė, 2014). In Europe, larvae of S. thuringiaca are leaf-miners of various herbaceous Rosaceae: Agrimonia, Filipendula, Fragaria, Potentilla and Sanguisorba. These plants also occur in the East Asiatic flora, but feeding preferenc- es of S. thuringiaca in East Asia are still unknown (Stonis & Rocienė, 2014). Formerly the species was known only from Europe (widespread from Spain to Central European Russia, and from Belgium to Italy and Ukraine; now it is recorded in deciduous, predominantly broad-leaf forest of the southern Primorye (Primorskiy Kray, Far East Russia). In addition, new data on poorly investigated species, namely Stigmel- la gimmonella (Matsumura), S. tranocrossa Kemperman & Wilkinson, and Ectoedemia philipi Puplesis, were disclosed. Uncus of S. gimmonella is divided into two gradually narrowed lateral lobes; bent distally in genitalia slide no. AG438 (Stonis & Rocienė, 2014: figs 36, 38). Gnathos with two slender caudal processes and large central plate. Valva with two apical processes bent inwardly and with heavily papillated inner lobe (Stonis & Rocienė, 2014: fig. 37). Transtilla with long sublateral processes. Cornuti spine-like, aggregated into long broad cluster, apical cornuti 2–3 times larger than the others. The male genitalia of S. gimmonella were illustrated/documented with photographs for the first time (Stonis & Rocienė, 2014). Transtilla of S. tranocrossa is with a large sublateral processes. Apical process of valva very large. Inner lobe of valva with extended and pointed corner. Cornuti vary in shape (Stonis & Rocienė, 2014). Central plate of gnathos of Ectoedemia philipi is distinctly serrate (Stonis & Rocienė, 2014: fig. 46). Valva with subbasal inner lobe, which was not documented/illustrated in the original description (Puplesis 1984b), nor visible in the holotype slide by Stonis & Rocienė 2013: figs

29 161, 162. Although the type of the species was studied recently, due to the bad quality of visibility, the subbasal inner lobe of the valva was not clearly visible and thus was not mentioned. The structure is reported by us for the first time (Stonis & Rocienė, 2014). The complete list of male specimens attracted to light during the fieldwork of 2011 in Primorskiy Kray comprises (see Stonis & Rocienė, 2014): Stigmella dissona (Klimesch): 11 males, 20.vii–5.viii.2011; S. sashai Puplesis: 182 males, 20.vii–11.viii.2011; S. sakhalinella Puplesis: 19 males, 26.vii–8.viii.2011; S. palionisi Puplesis: 2 males, 29.vii.2011; S. alisa Puple- sis: 3 males, 26.vii–5.viii.2011; S. auricularia Puplesis, Diškus & Juch- nevič: 4 males, 5–11.viii.2011; S. thuringiaca (Petry): 1 male, 6.viii.2011; S. tranocrossa Kemperman & Wilkinson: 3 males, 27.vii–11.viii.2011; S. sexcornuta Rocienė & Stonis (described in Rocienė & Stonis, 2014): 3 males, 6–11.viii.2011; S. gimmonella (Matsumura): 1 male, 5.viii.2011; S. dentatae Puplesis: 279 males, 19.vii–11.viii.2011; S. aladina Puplesis: 114 males, 19.vii–11.viii. 2011; S. omelkoi Puplesis: 1 male, 20.vii.2011; S. fervida Puplesis: 1 male, 5.viii.2011; S. multispicata Rocienė & Stonis (described in Rocienė & Stonis, 2014): 1 male, 8.viii.2011; Bohemannia manschurella Puplesis: 2 males, 27.vii.2011; B. nubila Puplesis: 48 males, 20.vii–11.viii.2011; Ectoedemia amani (Svensson): 37 males, 20.vii–11. viii.2011; E. admiranda Puplesis: 5 males, 7.viii–11.viii.2011; E. sivickisi Puplesis: 1 male, 26.vii.2011; E. arisi Puplesis: 4 males, 29.vii–8.viii.2011; E. christopheri Puplesis: 14 males, 19.vii–5.viii.2011; E. philipi Puplesis: 1 male, 19.vii.2011; E. chasanella Puplesis: 9 males, 26.vii–5.viii.2011; E. ortiva Rocienė & Stonis: 3 males, 19.vii–5.viii.2011; E. paraortiva Rocienė & Stonis (described in Rocienė & Stonis, 2014): 1 male, 5.viii.2011; E. olvina Puplesis: 7 males, 26.vii–5.viii. 2011; E. ornatella Puplesis: 18 males, 26.vii–7.viii.2011; E. ivinskisi Puplesis: 21 males, 19–29.vii.2011; Etainia tigrinella Puplesis: 2 males, 19.vii–11.viii.2011; and E. peterseni Puplesis: 2 males, 29.vii.2011 (Rocienė & Stonis, 2014). Although no attempt was made to assess which species of the East Asiatic fauna of Nepticulidae are common and which are rare, the abun- dance of the collected specimens may indicate that Stigmella sashai Puple- sis, S. dentatae Puplesis, S. aladina Puplesis, Bohemannia nubila Puplesis and Ectoedemia amani (Svensson) are among the most common species, at least in certain areas of Primorskiy Kray (Rocienė & Stonis, 2014).

30 After the fieldwork in northeastern China and dissection and identification of the collected material, we discovered some species whose distribution in the territory of China was known from other authors’ data (see van Nieukerken & Liu, 2000). Nepticulidae species discovered in continental East Asia that are new to science. This section presents 6 recently published (see Rocienė & Stonis, 2013; Stonis et al., 2013 and Stonis & Rocienė, 2014) new species which were discovered and described during the work on the dissertation. Most of them are trophically associated with plants of the Quercus genus. One of them, Stigmella cornuta Rocienė & Stonis (Stonis et al., 2013), is distributed in China and belongs to a newly described group S. cornuta, which is trophically associated with Quercus dentata. Another recently published new species, Stigmella multispicata Rocienė & Stonis (see Stonis & Rocienė, 2014) is described from the Russian Far East and belongs to the S. malella group; its host-plant is unknown. Other 4 newly described species are: Stigmella sexcornuta Rocienė & Stonis (see Stonis & Rocienė, 2014) belonging to the S. salicis species group (the host-plant unknown); Ectoedemia ortiva Rocienė & Stonis (see Rocienė & Stonis, 2013) and E. paraortiva Rocienė & Stonis (see Stonis & Rocienė, 2014) belonging to the E. suberis group (the species of this group are Quercus-miners); and Ectoedemia species 219 (see Rocienė & Stonis, 2013), which was also discovered and described but left unnamed (this species belongs to the subbimaculella group, which is trophically associated with Quercus). Stigmella cornuta Rocienė & Stonis (Figs 6–18) is a fuscous-winged species without fascia in the forewing. Some male genital characters (the apically narrowed valva, interrupted transverse bar of transtilla, broad lobate vinculum, and the uncus with the weakly individualized lobes) slightly resemble features of the species of the S. betulicola group, including the Oriental S. xystodes (Meyrick). However S. cornuta differs from all Stigmella species by the shape and number of the extremely large and pointed cornuti in the phallus and by the unusual shape of partially reduced caudal processes of gnathos. The combined leaf-mine (with a blotch-like second half) is also unique among Quercus-feeding Stigmella species (Stonis et al., 2013). Male forewing length is 1.9 mm; wingspan 4.2 mm. Head: palpi cream; frontal tuft beige-orange; collar and scape cream; antenna with 23 segments, shorther then half of

31 forewing length; flagellum fuscous brown on upper side and brownish cream on underside. Thorax: dorsum and tegulae concolorous with base of forewing. Forewing: mottled grey-brown, coarsely scaled (particularly in apical half), with weak greenish and purplish iridescence; fascia absent; cilia grey; underside of forewing uniformly fuscous brown. Hindwing and its cilia grey-brown, with no androconia. Legs brownish cream, with brown darkenings on foreleg upper side (Stonis et al., 2013). In the male genitalia, capsule longer (245 μm) than wide (170 μm). Vinculum with two short but broad lateral (anterior) lobes. Uncus with large, weakly individualized anterior lobes. Gnathos 85 μm broad, with two anterior and two posterior (caudal) processes. Valva 160 μm long, basally broad, narrowed apically, with small chitinized apical papilla; inner lobe absent; transtilla with interrupted transverse bar, without sublateral processes. Phallus 180 μm long, 60–70 μm broad; vesica with three extremely large pointed cornuti. Juxta membranous, narrowed medially. Manica absent. The host-plant is Quercus dentata Thunb. (Fa- gaceae) (Stonis et al., 2013). Egg flat and oval, on upper side of the leaf. Larvae mine in August. The first half of the mine is a narrow contorted gallery completely filled with black-green frass which turns brownish in old mines; in the second half of the mine gallery widens to an oval blotch with scattered black frass (Stonis et al., 2013). Larval exit slit on upper side of the leaf. Cocoon brownish cream; length 1.8 mm, maximal width 1.3 mm. Adults fly in September. It was found in deciduous broad-leaved forests of northeastern Chi- na. The species name is derived from Latin cornuta (horned) in reference to the extremely large cornuti of phallus. Stigmella multispicata Rocienė & Stonis (Figs 19–23) (recently des- ribed in Stonis & Rocienė, 2014) is diagnosed mostly on the basis of the number and shape of the cornuti, apically pointed valva, bilobed uncus and long lateral lobes of vinculum, the new species is attributed to the Stigmella malella species group (sensu lato, including the former Nearc- tic S. prunifoliella group). The male genitalia of S. multispicata resemble those of the Nearctic S. rhamnicola (Braun) and the predominantly East Mediterranean S. pyrellicola (Klimesch). However, from all known Stig- mella (including the species of the S. malella group), S. multispicata can be easily distinguished by the combination of very slender (process-like)

32 lateral lobes of the vinculum, subquadrate transtilla without sublateral processes, numerous flat, basally broadened, apically pointed cornuti, the apically purplish forewing with a shiny postmedian fascia, and the black frontal tuft on the head (Stonis & Rocienė, 2014). Male forewing length is about 2.1 mm; wingspan about 4.5 mm. Head: palpi cream; frontal tuft black; collar and scape cream with golden sheen; antenna with 19 segments, much shorter than half of forewing length; flagellum fuscous with bronzy sheen and greenish iridescence on upper side and underside. Tho- rax: dorsum and tegulae concolorous with base of forewing. Forewing narrow, area proximal to fascia shining coppery brown with slight purple tinge; fascia distinctly postmedian, shining silvery or brassy; distal to fascia violet to purple-brown; terminal cilia fuscous, shading into pale grey at tips; underside of forewing uniformly blackish brown. Hindwing unknown (lost in the holotype). Legs metallic grey, distally cream (Stonis & Rocienė, 2014). In the male genitalia, capsule 280 mm long. Vinculum with two very slender pointed lateral lobes. Uncus almost trapezioid, lateral lobes weakly individualized, rounded. Gnathos with two slender caudal processes and short slender transverse bar. Valva 190 mm long, gradually narrowed towards pointed apex; transtilla without sublateral processes. Juxta membranous but distictly chitinized at apical margin. Phallus broad (140 mm wide, 950 mm long); vesica with numerous flat pointed, basally broadened cornuti of different sizes (predominantly small in basal third, large in remaining two thirds) (Stonis & Rocienė, 2014). The host-plant is unknown. The only known specimen was collected in early August at light (Stonis & Rocienė, 2014). The type locality is in dense, deciduous mostly broad-leaf forests of Primorskiy Kray. The species name is derived from Latin multus/multi (numerous/numerously) and spicata (pointed/sharp), in reference to the numerous sharp cornuti and pointed sublateral lobes of the vinculum in the male genitalia (Stonis & Rocienė, 2014). Stigmella sexcornuta Rocienė & Stonis (Figs 24–28) (recently published in Stonis & Rocienė, 2014) belongs to the large and almost cosmo- politan Stigmella salicis species group (sensu lato, including the former Nearctic S. fuscotibiella group), in which species diagnosis sometimes is difficult and most often (but not only) relies on shape and number of -cor nuti. In male genitalia, S. sexcornuta most closely resembles the Japanese

33 S. azusa Hirano, a Salix serissaefolia feeding species recently described from Honshu, Japan (Hirano, 2010). However, from S. azusa and the remaining species of the S. salicis group, S. sexcornuta differs in the number and shape of cornuti: three cornuti are very broad and loose, three remaining are narrowed or very slender and form a cluster (Stonis & Ro- cienė, 2014). Male forewing length is 1.9–2.0 mm; wingspan 4.2–4.4 mm. Head: palpi cream; frontal tuft yellowish orange to orange; collar and scape cream to yellowish cream; antenna with 28 segments, almost half as long as forewing; flagellum grey-brown on upper side and pale brownish to brown on underside. Thorax: dorsum and tegulae concolorous with base of forewing. Forewing: area proximal to fascia mottled grey brown, sometimes with coppery sheen; fascia postmedial, whitish to shining silvery, slightly broadened at tornus; distal to fascia coarsely scaled (mottled with blackish grey), with weak purplish iridescence; terminal cilia grey; underside of forewing uniformly dark grey-brown, with weak purplish sheen. Hindwing and its cilia grey, with no androconia. Legs ochre-grey to brown, glossy, with blackish or brown darkenings on tarsi (or foreleg tibia and tarsi) (Stonis & Rocienė, 2014). In the male genitalia, capsule 250 mm long. Vinculum with two short triangular lobes and broad trap- ezoidal anterior excavation. Uncus gradually narrowed caudally, with four small caudal lobes. Gnathos (65 mm) with large lateral arms and two parallel long and slender caudal processes. Valva 170 mm long, with broad angular inner lobe and short pointed, slightly inwardly-bent apical process; transtilla without sublateral processes. Phallus broad (80 mm wide, 215 mm long); vesica with six cornuti: three very broad and loose; remaining three narrowed or very slender and aggregated into a cluster (see also Stonis & Rocienė, 2014: figs 20, 21). The host-plant possibly is Salix sp. Adults fly in August, collected at light. The type locality is in dense, deciduous mostly broad-leaf forests of Primorskiy Kray. The species name is derived from Latin sex (six) and cornuti (horns/cornuti) in reference to the characteristic/diagnostic number of six cornuti on the vesica in the phallus of the male genitalia (Stonis & Rocienė, 2014). Ectoedemia ortiva Rocienė & Stonis (Figs 31, 32, 34, 38) belongs to the Ectoedemia suberis group. The male is recognized by the fuscous grey-brown hindwing with yellowish cream lamellar androconial scales surrounding the yellowish cream hair-pencil. Male genitalia are char-

34 acterized by the shape of a large curved valva without the inner margin convex (in the majority species of the group, the valva is distinctly broad at basal part, with inner marging convex, and narrowed below the middle) (Rocienė & Stonis, 2013). This recently described species resembles the predominantly Sub-Mediterranean E. caradjai (Groschke) and the West Mediterranean E. phaeolepis van Nieukerken, A. Laštuvka & Z. Laštuvka and E. coscoja van Nieukerken, A. Laštuvka & Z. Laštu- vka (see van Nieukerken et al., 2010), but E. ortiva differs by its broad phallus with numerous spine-like cornuti and a dark hindwing with a yellowish cream patch of androconia and cream hair-pencil (the costal margin of the hindwing is also cream) (Rocienė & Stonis, 2013). Male forewing length is 2.6 mm; wingspan 5.6 mm. Head: palpi brownish cream; frontal tuft orange; collar cream comprising piliform scales; scape yellowish-cream; antenna with 28 segments, shorter than half forewing length; flagellum brown on upper side and underside. Thorax: dorsum and tegulae grey-brown. Forewing grey-brown, with distinct bronze lus- tre, apically speckled with some fuscous brown scales; median fascia not prominent, grey-white or silver-white (depending on the angle of view); cilia pale brown; underside of forewing uniformly dark brown. Hind- wing broadened at basal half, fuscous or grey-brown on upper side and underside; hair-pencil not prominent, grey-cream or cream, surrounded by a large patch of yellowish cream androconia; costal margin yellowish cream; cilia dark brown. Legs brown-cream. The male genitalia with capsule just slightly longer (288 mm) than wide (249 mm). Vinculum with two very short but broad lateral lobes. Pseuduncus triangular. Gnathos with long lateral arms and caudal process. Valva slender (58 mm), 178 mm long, curved inwardly, gradually narrowed towards apex, with pointed apical process; transtilla with prominent sublateral processes. Phallus broad (124 mm), 284 mm long, with two apical carinae; vesica with numerous spine-like and denticulate cornuti of different size (Rocienė & Stonis, 2013). E. ortiva possibly is a leaf-miner on Quercus. Adults fly in July, attracted to light. It occurs in dense, mostly deciduous forests of Pri- morskiy Kray. This species is named after the region East Asia – the Latin “ortivus” means the eastern (dawn) side (Rocienė & Stonis, 2013). Ectoedemia paraortiva Rocienė & Stonis (Figs 29, 30, 33, 35, 36, 37) (recently published by Stonis & Rocienė, 2014) belongs to the Ectoe-

35 demia suberis group and to most closely resembles the other recently described East Asiatic species, E. ortiva Rocienė & Stonis (2013). Both species (E. ortiva and E. paraortiva) differ from other Ectodemia by the broad phallus with numerous spine-like cornuti (Stonis & Rocienė, 2014). E. paraortiva differs from E. ortiva by the absence of a fascia on the forewing (in E. ortiva the fascia is well developed), also by the more grey hindwing with a long, weakly visible hair-pencil, which is not surrounded by a yellowish cream patch of androconia (in E. ortiva this patch is very characteristic). Male forewing length is about 2.0 mm; wingspan about 4.4 mm. Head: palpi grey-cream; frontal tuft pale orange; collar whitish, comprising loose lamellar scales; scape cream; antenna with 30 segments, almost half as long as forewing; flagellum grey on upper side and underside. Thorax: dorsum and tegulae concolorous with base of forewing. Forewing grey-brown, apically more coarsely speckled with darker scales; fascia absent; cilia pale grey-brown; underside of forewing uniformly dark grey-brown. Hindwing brownish grey or dark grey on upper side and underside; hair-pencil slender and long (longer than half of hindwing), but not prominent (hardly visible), grey-cream (but not surrounded by a patch of cream androconia as in E. ortiva); costal margin remains brownish grey; cilia pale grey to grey. Legs brown-cream, shaded dark brown on upper side. In the male genitalia, capsule longer (270 mm) than wide (180 mm). Vinculum very broad (180 mm), with two very short but very broad lateral lobes. Pseuduncus triangular, damaged in genitalia slide no. AG428 (Stonis & Rocienė, 2014). Gnathos with long lateral arms and slender caudal process. Valva 170 mm long, curved inwardly, gradually narrowed towards apex, with pointed apical process; transtilla with slender and long sublateral processes. Phallus broad (100 mm), 220 mm long, with two apical carinae; vesica with numerous spine-like or similar cornuti of different sizes. Although the phallus of E. paraortiva tends to be slightly smaller than that of E. ortiva, E. paraortiva is not distinguishable on basis of the phallus (see Stonis & Rocienė, 2014: figs 30, 31). The species is possibly a leaf-miner onQuercus (as the other species of the Ectoedemia suberis group). The only known specimen was collected in early August, attracted to light. The type locality is in dense, deciduous mostly broad-leaf forests of Primorskiy Kray. This species is named after Ectoedemia ortiva Rocienė & Stonis, a closely sim-

36 ilar (and presumably closely related) species recently described from the same locality in Primorskiy Kray as the new species (see Rocienė & Sto- nis 2013). We suppose that E. ortiva and E. paraortiva are probably two sympatric species (Stonis & Rocienė, 2014). Ectoedemia species 219 (Figs 39–44) belongs to the Ectoedemia subbimaculella group. The male is recognized by a combination of the grey-white fascia of forewing, a weakly developed brownish hair-pencil surrounded by a patch of whitish androconia, and features of the male genitalia (broad valva, weakly developed central element of gnathos, very short vinculum) (Rocienė & Stonis, 2013). Male forewing length is 2.5 mm; wingspan 5.5 mm. Head: palpi brownish cream; frontal tuft orange; collar orange-cream, comprising piliform scales; scape orange-cream; antenna with 35–36 segments, slightly shorter than half forewing length; flagellum pale brown on upper side, cream to pale brown on underside. Thorax: dorsum and tegulae pale brown. Forewing pale grey-brown, speckled with dark grey-brown and fuscous scales, with oblique grey-white median fascia; cilia pale brown; underside uniformly pale brown to dark brown (depending on angle of view). Hindwing pale grey-brown on upper side and underside; hair-pencil very weakly developed, comprises a few pale grey-brown to dark brown setae, surrounded by a patch of white or grey-white androconia; cilia pale grey-brown. Legs pale brown, glossy. The male genitalia with capsule considerably longer (256 mm) than wide (194 mm). Ventral plate of vinculum very short, lateral lobes absent. Pseuduncus triangular. Gnathos with long narrow lateral arms and weakly developed central element (no caudal process). Valva 199 mm long, broadened medially, with strongly slerotized basal margin and short apical process; transtilla with long slender transverse bar and short slender sublateral processes. Phallus about 263 mm long (possibly partially everted in Rocienė & Stonis, 2013: figs 78, 79) and relatively slender (66–77 mm); vesica thickened laterally and with numerous minute cornuti. The species is possibly a leaf-miner on Quercus. Adults fly in July, attracted to light. It occurs in dense, mostly deciduous forests of Primor- skiy Kray. This new species left unnamed because of the poor quality of the material (Rocienė & Stonis, 2013).

37 FIGURES 6 –6–12.12. Stigmella Stigmella cornuta cornutaRocienė Rocienė & Stonis & Stonis,, a recently a recently described describedspecies. species.6 – habitat, deciduous6 – habitat, broad deciduous-leaf forests broad-leaf in northeastern forests China, in northeasternHuairou Distric t,China, 40°25'56''N, Huairou 116°33'20''E; District, 8 – cocoon,40°25’56’’N, scale bar 116°33’20’’E; 1 mm; 7, 9–12 8, leaf– cocoon,-mines on scaleQuercus bardentata 1 mm;Thunb., 7, 9–12, Fagaceae leaf-mines (arrows indicate on the narrow part of the mines: a gallery completely filled with black-green or brownish frass) (publishedQuercus bydentata the author Thunb., in a co- authoredFagaceae paper (arrows by Stonis indicate et al., 2014)the narrow. part of the mines: a gallery completely filled with black-green or brownish frass) (published by the author in a co-authored paper by Stonis et al., 2014). 24

38 FIGURES 13 –13–18.18. Stigmella Stigmella cornuta Rocienėcornuta & SRocienėtonis, a recently & Stonis, describ aed recently species. 13described – male adult, holotype;species. 1314 – same,male forewing,adult, holotype; artist reconstruction, 14 – same, scale forewing, bar 1 mm; artist 15 reconstruction,– male genitalia, holotype,scale gen.bar slide1 mm; no. 15 AG419, – male scale genitalia, bar 100 holotype, μm; 16 – same, gen. transtillaslide no.; 17AG419, – same, scale vinculum bar 100(const μm;ricted laterally); 18 – valva and juxta (the arrow indicates a chitinized apical papilla). Scale bar 50 μm (published16 – same, by thetranstilla; author in 17a co –- authoredsame, vinculum paper by Stonis (constricted et al., 2014). laterally); 18 – valva and juxta (the arrow indicates a chitinized apical papilla). Scale bar 50 μm (published by the author in a co-authored paper by Stonis et al., 2014). 25

39 Figures 19–23. Stigmella multispicata Rocienė & Stonis, a recently described species,Figures holotype, 19–23. Stigmella genitalia multispicata slide no. AG427Rocienė (ZIN): & Stonis, 19 –a adult;recently 20 described – forewing, species, artistholotype, genitalia slide no. AG427 (ZIN): 19 – adult; 20 – forewing, artist reconstruction, scale bar 1 mm; 21 reconstruction,– male genitalia, scale capsule; bar 22 1 – mm;same, 21uncu – s,male gnathos, genitalia, valva and capsule; juxta; 23 22 – phallus – same,. Scale uncus, bar 100 µm gnathos,(published valva by theand autho juxta;r in 23a co –- authoredphallus. paper Scale by bar Stonis 100 & µmRocienė, (published 2014). by the author in a co-authored paper by Stonis & Rocienė, 2014).

Figures 24–28. Stigmella sexcornuta Rocienė & Stonis, a recently described species:Figures 2424–28 –. adult,Stigmella holotype; sexcornuta 25Rocienė – same, & paratype. Stonis, a recently Scale describedbar 1 mm; species: 26 –24 male– adult, genitalia,holotype; 25 capsule, – same, paratype. holotype, Scale genitalia bar 1 mm; slide 26 – maleno. genitalia,AG429 (ZIN);capsule, 27holotype, – same, geni taliawith slide no. AG429 (ZIN); 27 – same, with valvae removed, paratype, genitalia slide no. AG431 (ZIN); 28 – valvaevalvae andremoved, phallus, paratype, paratype, genitalia genitalia slide slide no. no. AG430 AG431 (ZIN) (ZIN);. Scale 28 bar – 100valvae µm and(published phallus, by the paratype,author in a cogenitalia-authored paperslide byno. Stonis AG430 & Rocienė, (ZIN). 2014). Scale bar 100 µm (published by the author in a co-authored paper by Stonis & Rocienė, 2014). 27

41 FiguresFigures 2929–34.–34. Two Two poss possiblyibly sympatric sympatric Ectoedemia Ectoedemia species (after species Stonis & (afterRocienė, Stonis 2014): & 29 – E. Rocienė,paraortiva 2014):Rocienė 29 –& E.Stonis, paraortiva holotype ,Rocienė adult (ZIN); & Stonis,30 – same, holotype, hindwing; adult 31 – E.(ZIN);ortiva 30Rocienė – & Stonis, holotype, adult (ZIN); 32 – same, hindwing. Scale bar 1 mm. 33 – male genitalia, capsule, E. same,para hindwing;ortiva Rocienė 31 – & E. Stonis, ortivaholotype, Rocienė genitalia & Stonis,slide holotype, no. AG428 adult (ZIN); (ZIN); 34 32– same,– same, E. ortiva hindwing.Rocienė & Scale Stonis, bar a not1 mm. type 33series – malespecimen, genitalia, genitalia capsule, slide no. E. AG207 paraortiva (ZIN). RocienėScale bar &100 µm Stonis,(published holotype, by the genitaliaauthor in co slide-authored no. AG428papers by (ZIN); Rocienė 34 & – Stonis, same, 2013 E. ortivaand Stonis Rocienė & Rocienė, & 2014).Stonis, a not type series specimen, genitalia slide no. AG207 (ZIN). Scale bar 100 µm (published by the author in co-authored papers by Rocienė & Stonis, 28 2013 and Stonis & Rocienė, 2014).

42 .

Figures 35–38. Details of male genitalia of two possibly sympatric Ectoedemia species. 35 – E. Figuresparaortiva 35–38.Rocienė Details& Stonis of(published male genitalia in Stonis of& Rocienė,two possibly 2014), sympatricmale genital Ectoedemiacapsule, holotype species.(reconstruction), 35 – E. genparaortivaitalia slide Rocienė no. AG428 & Stonis(ZIN); (published36 – same, valva;in Stonis 37 – &phallus Rocienė, (with 2014), basal part maletorn), genitalE. para ortivacapsule, Rocienė holotype & Stonis, (reconstruction), holotype, genitalia genitalia slide no. slide AG428 no. (ZIN); AG428 38 (ZIN);– same, E. ortiva Rocienė & Stonis (published in Stonis & Rocienė, 2014), a non type series specimen, genitalia 36 –slide no.same, AG433 valva; (ZIN). 37 Scale – phallus bar 100 µm.(with basal part torn), E. paraortiva Rocienė & Stonis, holotype, genitalia slide no. AG428 (ZIN); 38 – same, E. ortiva Rocienė & Stonis (published in Stonis & Rocienė, 2014), a non type series specimen, genitalia slide no. AG433 (ZIN). Scale bar 100 µm. 29

43 FIGURESFIGURES 39 39–44.–44. Ectoedemia Ectoedemia species species 219, slide 219, no. slide AG219 no. (ZIN). AG219 39 –(ZIN).adult, male.39 – Scaleadult, bar 1 mm; 40 – valva; 41 – capsule (phallus removed); 42 – same, reconstruction; 43 – phallus; 44 – same, male.reconstruction. Scale bar Scale 1 barmm; 100 40 μm –(recently valva; 41published – capsule by the (phallus author in aremoved); co-authored 42paper – bysame, Rocienė reconstruction;& Stonis, 2013). 43 – phallus; 44 – same, reconstruction. Scale bar 100 μm (recently published by the author in a co-authored paper by Rocienė & Stonis, 2013).

44 Results of the taxonomic analysis. The most intensive studies of the East Asiatic Nepticulidae took place in 1984–1987. During that period, over 70 new Nepticulidae species were described. The recent 15-year period was not very productive for faunistic studies of the East Asiatic Nepticulidae. It was only in 2010 that some additions to the fauna were introduced (see Literature Overview). The results of our investigations combined with other authors’ data have yielded an updated and annotated checklist of the Nepticulidae of East Asia. In addition, the author updated the data on distribution, morphology and diagnosis of many examined species. After the addition to the East Asiatic fauna of the formerly European Stigmella thuringiaca (Petry) and S. cornuta Rocienė & Stonis from Chi- na (Stonis et al., 2013), and the recent description of Stigmella multispicata Rocienė & Stonis, S. sexcornuta Rocienė & Stonis, and Ectoedemia paraortiva Rocienė & Stonis (Stonis & Rocienė, 2014), in total the current checklist of the Nepticulidae of East Asia comprises 110 described and named species, and one described but unnamed species (Ectoedemia species 219 documented by Rocienė & Stonis, 2013). Seventy-one species occur in the Russian Far East and 53 in Japan (20 species of which occur both in Japan and Russia). Of the 53 species currently known from Ja- pan, six species (approximately 11%) also occur in Europe. For 12 species (about 17%) known from the continental Russian Far East (Primorskiy Kray), the Euro-East Asiatic distribution is characteristic. All species identified in East Asia belong to five genera: Stigmella Schrank, Bohemannia Stainton, Ectoedemia Busck, Fomoria Beirne, and Etainia Beirne. The Stigmella genus comprises the largest number of species (69); 27 species belong to Ectoedemia, 6 to Bohemannia, 5 to Etainia, and 3 to Fomoria (Fig. 45). During the dissertation period the taxonomic checklist of the East Asiatic Nepticulidae changed because some species names were synonymized. Stigmella regina was determined to be a junior synonym of S. sashai, and Ectoedemia laura of E. sivickisi (Rocienė & Stonis, 2013). Results of the chorological analysis. All currently known East Asiatic Nepticulidae species were mapped (with global distribution in- dicated) and grouped according to the similarity of ranges (species with similar or identical ranges were merged into one chorological group). The

45 Fig. 45. Taxonomic composition of the Nepticulidae of East Asia. analysis of distribution ranges showed that chorologically the East Asiat- ic fauna of Nepticulidae is not homogeneous. Four chorological groups of Nepticulidae can be distinguished: the East-Palaearctic (we further divide it into continental, insular, and broad), Amphi-Palaearctic, Hol- arctic, and the Trans-Palaearctic. East Asiatic Nepticulidae species are characterized by both very broad (Holarctic or Palearctic) distribution ranges and geographically more limited ranges. Th e smallest chorological groups of East Asiatic fauna were found to be groups with the broadest distribution ranges, i.e. Holarctic and Amphi-Palaearctic comprising 1–2 species each. Th e most abundant chorological group of the East Asiatic fauna is the Continental East-Palaearctic group but not the Broad East-Palaearctic as predicted earlier (Fig. 46). It is interesting that most of theTrans-Palaearctic species were recorded in the fauna of Japan.

46 Fig. 46. Chorological composition of the East Asiatic fauna of Nepticulidae.

The East-Palaearctic chorological group. The species of this group are distributed in East Asia: their distribution ranges extend from the northeastern provinces of China, Primorskiy Kray, Sakhalin, North Korea and South Korea up to Japan (most often Hokkaido Island). In the East Asiatic fauna, this chorological group comprises 91 Nepticulidae species (83%). According to the species distribution, the East Palaearctic chorological group can be further subdivided into the Continental (continental part of Asia), the Insular (Japan and the neighbouring islands) and the Broad (i.e. the continental part of East Asia and Japan together) chorological subgroups (Rocienė & Stonis, 2014). The Continental East-Palaearctic chorological subgroup. The great- est species diversity (it comprises 43 Nepticulidae species) (Rocienė & Stonis, 2014): Stigmella mirabella (Pupl.), S. sashai Pupl., S. attenuata Pupl., S. ultima Pupl., S. tegmentosella Pupl., S. kozlovi Pupl., S. nostrata Pupl., S. micromelis Pupl., S. crataegivora Pupl., S. taigae Pupl., S. palionisi Pupl., S. amuriella Pupl., S. alisa Pupl., S. auricularia Pupl., Diškus & Juchn., S. sexcornuta Rocienė & Stonis, S. palmatae Pupl., S. monticulella Pupl., S. fervida Pupl., S. multispicata Rocienė & Stonis, S. cornuta Ro- cienė & Stonis, Bohemannia suiphunella Pupl., B. piotra Pupl., Ectoedem- ia admiranda Pupl., E. sivickisi Pupl., E. arisi Pupl., E. christopheri Pupl.,

47 E. philipi Pupl., E. chasanella Pupl., E. ortiva Rocienė & Stonis, E. paraortiva Rocienė & Stonis, E. aligera Pupl., E. ermolaevi Pupl., E. maculata Pupl., E. species 219, E. ornatella Pupl., E. ivinskisi Pupl., E. picturata Pupl., E. sinevi Pupl., Fomoria permira Pupl., Etainia capesella (Pupl.), E. tigrinella (Pupl.), E. peterseni (Pupl.), and E. sabina (Pupl.). The species of this subgroup have a wide spectrum of host-plants, but mainly they are trophically associated with plants of the genera Acer, Quercus, and Betula (Rocienė & Stonis, 2014). The Insular East-Palaearctic subgroup comprises 30 species: Stigmella caesurifasciella Kemp. & Wilk., S. kurilensis Pupl., S. conchyliata Kemp. & Wilk., S. oplismeniella Kemp. & Wilk., S. populnea Kemp. & Wilk., S. titivillitia Kemp. & Wilk., S. orientalis Kemp. & Wilk., S. alaurulenta Kemp. & Wilk., S. chaenomelae Kemp. & Wilk., S. honshui Kemp. & Wilk., S. sorbivora Kemp. & Wilk., S. zumii Kemp. & Wilk., S. kurot- subarai Kemp. & Wilk., S. nakamurai Kemp. & Wilk., S. nireae Kemp. & Wilk., S. azusa Hirano, S. zelkoviella Kemp. & Wilk., S. acrochaetia Kemp. & Wilk., S. alikurokoi Kemp. & Wilk., S. ichigoiella Kemp. & Wilk., S. sesplicata Kemp. & Wilk., S. spiculifera Kemp. & Wilk., S. oa Kemp. & Wilk., S. crenatiella Hirano, S. azuminoensis Hirano, S. hisakoae Hirano, Bohemannia nipponicella Hirano, Ectoedemia cerviparadisico- la Sato, E. insularis Pupl., and Etainia trifasciata (Mats.) (Rocienė & Sto- nis, 2014). In terms of trophic relationships, the larvae of this group mine plants belonging to 15 genera: Acer, Alnus, Quercus, Populus, Malus, Sorbus, Rhamnus, Ulmus, Salix, Rubus, Rhododendron, Castanea, Oplis- menus, Zelkova, and Chaenomeles (Rocienė & Stonis, 2014). The Broad East-Palaearctic chorological subgroup includes 18 Nep- ticulidae species: Stigmella cathepostis Kemp. & Wilk., S. monella Pupl., S. tranocrossa Kemp. & Wilk., S. gimmonella (Mats.), S. dentatae Pupl., S. aladina Pupl., S. omelkoi Pupl., S. fumida Kemp. & Wilk., S. clisio- tophora Kemp. & Wilk., S. castanopsiella (Kuroko), S. kurokoi Pupl., Bohemannia manschurella Pupl., B. ussuriella Pupl., B. nubila Pupl., Ectoedemia scoblei Pupl., E. olvina Pupl., E. pilosae Pupl., and Fomoria hypericifolia Kuroko. The species of this group have a wide spectrum of host-plants, yet they are mostly trophically associated with plants of Quercus and Acer genera (Rocienė & Stonis, 2014) (Fig. 47).

48 Fig. 47. Trophic relationships of the species of the Broad East-Palaearctic chorological subgroup in the fauna of East Asia.

Th e Amphi-Palaearctic chorological group. Th e species of this group are characterized by a disjunct distribution range, one part of which is in the southern or south-eastern part of Europe, including Italy, Slovakia, Czech Republic, Austria, Hungary, and Greece; the other is in East Asia (Russian Far East, northeastern China, and Korea). Th e fauna of East Asia has one species belonging to this chorological group, namely Ectoe- demia preisseckeri (Klim.), which is trophically associated only with the Ulmus genus. Th e Holarctic chorological group. Th is chorological group encompasses species that have the broadest geographical distribution. Th e ranges of species extend from Europe to East Asia and North America. Th e fauna of East Asia has two species belonging to this chorological group: Ectoedemia argyropeza (Z.) and E. occultella (L.). Th e fi rst is trophically associated with plants of the genus Betula, the other with Populus (Fig. 48). Th ough the leaf-mining Ectoedemia argyropeza may have been introduced into North America accidentally (with host-plants), we have no evidence to confi rm or deny this assumption so far (Rocienė & Stonis, 2014).

49 Betula Populus 50% 50%

Fig. 48.Trophic relationships of the species of the Holarctic chorological group in the fauna of East Asia. 376 pav. Holarktinio paplitimo rūšių trofiniai ryšiai Rytų Azijos faunoje.

The Trans-Palaearctic chorological group. This group encompasses species with the rangesTranspalearktinė extending from chorologinė Europe (through grupė southern Siberia) to East Asia: Russian Far East (Sakhalin and Primorskiy Kray), northern provinces of China and Japan (most often Hokkaido Island). Though we Šiai rūšių grupei priklauso rūšys, kurių arealai driekiasi nuo Europos (per pietinį Sibirą) iki still have no sufficient evidence on the continuous range of the so-called Rytų Azijos: Trans-Palaearctic Rusijos Tolimųjų Rytųspecies, (Sachalino the host-plants ir Primorės of these krašto), species šiaurinių do Kinijoshave such ir Japonijos provincijų (dažniausiaidistribution Hokaido ranges. salos).This could Nors ikibe šiolone visof the dar main trūksta arguments įrodymų allowing apie vadinamųjų transpalearktiniųthese rūšių species ištisinį to be arealą, called tačiau Trans-Palaearctic šių rūšių mitybiniams but not Amphi-Palaearctic augalams toks arealas yra (which are characterized by a large gap of the range in the centre of the būdingas. Tai gali būti vienas svarbiausių argumentų, leidžiančių šias rūšis įvardinti kaip continent). Currently the Trans-Palaearctic group includes 16 Neptic- transpalearktines, o ne amfipalearktines (kurioms būtų būdingas didelis arealo trūkis žemyno centre). ulidae species of the East Asiatic fauna (Rocienė & Stonis, 2014). The Transpalearktineimajority rūšių of grupei species šiuo are metu trophically priskiriam associateda 16 Rytų with Azijos Betulaceae: faunos NepticulidaeStigmella rūšių. Dauguma šių naturnellarūšių trofiškai (Klim.), yra susijusios S. betulicola su Betulaceae: (Stt.), S. Stigmella lutella (Stt.),naturnella S. sakhalinella(Klim.), S. betulicola (Stt.), S. lutellaPupl., (Stt.), S. continuella S. sakhalinella (Stt.),Pupl., Salicaceae: S. continuella S. assimilella(Stt.), Salicaceae: (Z.), S. salicisS. assimilella (Stt.), (Z.), S. S. abliquella (Hein.), E. intimella (Z.), and E. hannoverella (Glitz); other salicis (Stt.), S. abliquella (Hein.), E. intimella (Z.), E. hannoverella (Glitz); kitos rūšys susijusios su species are associated with plants of Rosaceae (S. aurora Pupl., S. anom- Rosaceae (S. aurora Pupl., S. anomalella (Göze), S. thuringiaca (Petry)), Ericaceae (S. lediella alella (Göze), S. thuringiaca (Petry)), Ericaceae (S. lediella (Schl.), Fomoria (Schl.), Fomoriaweaveri weaveri (Stt.)),(Stt.)), and Ulmaceaeir Ulmaceae (Ectoedemia (Ectoedemia amani amani (Svenss.))(Svens.)) (Rocienė šeimų augalais & (377 pav.). ChorologiniuStonis, požiūriu 2014) (Fig. viena 49). tipiškiausių Chorologically, šios grupės Stigmella rūšių yra anomalella labai plataus is one paplitimo of the Stigmella anomalella, most kuri paplitusitypical species nuo Rusijosof this group, Tolimųjų which Rytų has iki a very Europos: broad Didžiosiosdistribution Britanijos, Portugalijos, fromSuomijos, the Russian Prancūzijos Far beiEast Kanarų to Europe: salų. BeneGreat pats Britain, įdomiausias Portugal, yra Finland,Stigmella lediella arealas, kuris apima tik šiaurrytinę Europos dalį ir, matyt, nepertraukiamai tęsiasi per Sibirą iki 50 Rusijos Tolimųjų Rytų. Europoje ši rūšis minuoja tik Ledum, o Rytų Azijoje – tik įvairius

178 France, and the Canary Islands. Probably the most peculiar is the range of Stigmella lediella. It covers only the northeastern part of Europe and, most likely, continuously extends through Siberia up to the Russian Far East. In Europe this species is mining only Ledum, and in East Asia only various Rhododendron plants (Rocienė & Stonis, 2014). Earlier this East Asiatic population was described as two species diff ering from the European S. lediella (Puplesis, 1985b); however, later it was proved that their genital structures are identical, and thus the names of both Asiatic species were recognized to be junior synonyms of S. lediella (Puplesis & Diškus, 2003). The highest diversity of the Nepticulidae species (and endemism) was recorded in the Russian Primorskiy Kray, which is characterized by exceptional biodiversity. Primorskiy Kray is recorded to be inhabited by 69 Nepticulidae species, 57 of which (82%) currently are known as endemic in Primorskiy Kray (Rocienė & Stonis, 2014). Th ough the overall level of endemism is rather high, 16 species (15%) detected in continental East Asia are distinguished by very broad ranges. Ectoedemia occultella (L.) and E. argyropeza (Z.) are characterized by Holarctic distribution, and the ranges of other 14 species extend up to Europe (Rocienė & Stonis, 2014). Disclosing the fauna of Siberia might reduce the number of endemic species; however, this enormous region of Asia is still insuffi ciently studied as regards Nepticulidae.

Fig. 49. Trophic relationships of the species of the Trans-Palaearctic chorological group in the fauna of East Asia.

51 The chorological analysis revealed 7 possible vicariants: Stigmella sexcornuta Rocienė & Stonis (Primorskiy Kray) is probably vicariant with S. azusa Hirano (Japan), S. fervida Pupl. (Primorskiy Kray) with S. hisakoae Hirano (Japan), S. nostrata Pupl. (Primorskiy Kray) with S. zumii Kemp. & Wilk. (Japan), S. palionisi Pupl. (Primorskiy Kray) with S. nakamurai Kemp. & Wilk. (Japan), S. palmatae Pupl. (Primorskiy Kray) with S. filipendulae(Wocke) (Europe), Bohemannia ussuriella Pupl. (Primorskiy Kray) with B. nipponicella Hirano (Japan), and B. piotra Pupl. (Primorskiy Kray) with B. pulverosella (Stt.) (Europe) (Rocienė & Stonis, 2014). The Stigmella Schrank species groups trophically associated with Quercus as a host-plant. Since species groups in Nepticulidae are highly valuable for diagnostic purposes to make orientation easier, especially in such big genera as Stigmella with about 400 species described worldwide, in present dissertation we recognized most of the groups listed below: the caerusifasciella, the cornuta, the saginella, the quercipulchella, the ruficapitella, the castanopsiella and the hemargyrella groups (published by the author in a co-authored paper by Stonis et al., 2013). The first species groups of Stigmella associated with Quercus, the ruficapitella and the hemargyrella groups, were proposed for European species on the basis of rather uncommon male genitalia and feeding pref- erences (Johansson, 1971). Later, in the revisions of the NW European or West Palaearctic Nepticulidae, Johansson et al. (1990) and van Nieukerk- en & Johansson (2003) combined the hemargyrella and the ruficapitella groups into one, the ruficapitella group, because monophyly of the hemargyrella group seemed doubtful (van Nieukerken & Johansson, 2003). If initially the ruficapitella group contained only one non-Quercus-feeding species (Betula-feeder S. tristis Wocke), now the enlarged group comprised a few non-Quercus feeding species, including the European S. hemargyrella, S. speciosa and S. lonicerarum feeding on Fagus, Acer or Lonicera, respectively (Johansson et al., 1990; van Nieukerken & Johans- son, 2003). Because of some differences in the genitalia, the merge of two groups was not followed in the world catalogue (Diškus & Puplesis, 2003) (published by the author in co-authored paper by Stonis et al., 2013). Revising the Canadian fauna, Wilkinson & Scoble (1979) recognized two other species groups: the saginella group and the quercipulchella

52 group. Later, the caesurifasciella group was designated by Kemperman and Wilkinson (1985) for the East Asiatic Quercus-feeding Stigmella species. In the same publication by Kemperman & Wilkinson (1985), the suberivora species group was erected. However, it was not recognized by any other authors, and its species currently are included in the ruficapitella group (van Nieukerken & Liu, 2000; van Nieukerken & Johansson, 2003; Diškus & Puplesis, 2003). Reviewing Asiatic Nepticulidae, Puplesis (1994) removed two species with a bulbous phallus and a coiled vesica from the ruficapitella group and erected a separate group for these, the castanopsiella group (Puplesis, 1994). Later, in a paper mainly on Chi- nese Stigmella feeding on Quercus, the castanopsiella group was only partially recognized (van Nieukerken & Liu, 2000). According to these authors, the castanopsiella group most likely constitutes a monophyletic entity, but separation from the ruficapitella group would probably render the remainder paraphyletic (van Nieukerken & Liu, 2000). Thus, until a thorough phylogenetic analysis has been carried out, these authors (van Nieukerken & Liu, 2000) preferred to keep the castanopsiella group in the wide sense (as part of the ruficapitella group) (published by the author in co-authored paper by Stonis et al., 2013). Designation of the Stigmella cornuta group, and cladistic analysis. Designation of the cornuta species group is based on Stigmella cornuta, a species that we recently discovered in China (published by the author in a co-authored paper by Stonis et al., 2013). This new species group can be recognized by the following diagnostic characters: the forewing without fascia; in the male genitalia, uncus with weakly individualized lobes, gnathos with two partialy reduced caudal processes and two anterior ones, valva with chitinized apical papilla, tranverse bar of transtilla interrupted medially, phallus with extremely large pointed cornuti, juxta present, constricted medially, manica absent. The combined leaf-mine (Figs 9, 10) is also unique among Quercus-feeding Stigmella species (and uncommon or even rare among other Nepticu- lidae): comprise a narrow gallery in the first half and a large blotch in the second half (unique among other Stigmella species feeding on oak) (published by the author in co-authored paper by Stonis et al., 2013). In the male genitalia, the cornuta group exibit some similarity to the non Quercus-feeding betulicola group: the apically narrowed valva

53 with a chitinized apical papilla, the interrupted transverse bar of transtilla, the broad lobate vinculum, and the weakly individualized lobes of uncus (Stonis et al., 2013; Rocienė & Stonis, 2014). A cladistic analysis of the species of the Stigmella betulicola group and S. cornuta detected in East Asia showed that S. cornuta does not belong to the S. betulicola group but is a representative of a separate (independent) group of species. The difference and, in terms of evolution, early separation of S. cornuta is demonstrated by the right clade in a cladogram, which is based on 9 autapomorphic characters (Fig. 50). The most important of them are the following: the development of very large cornuti in phallus, trophic specialization (Fagaceae: Quercus mining), and the mine type (combined leaf-mines). The monophyly of the S. betulicola group is evidenced by 7 autapomorphic characters, the main of which are the following: the apical sclerotization of valva turned into a papilla, the total reduction of cornuti (phallus without cornuti), and the development of specialized paired carinae (Fig. 50).

A list of apomorphic characters based on which the status of the cornuta group was proved: 1. Transverse bar of transtilla interrupted 2. Valva narrowed in the apical part 3. Development of wide lobes of vinculum; only occasionally vinculum rounded 4. Development of apical sclerotization of valva 5. Transtilla rounded laterally 6. Development of U-shape gnathos* 7. Vinculum shortened* 8. Adult forewing without a fascia (fascia reduced) 9. Trophic specialization: Fagaceae – Quercus mining* 10. Mine specialization: the combined mines* 11. Apical sclerotization of valva turned into a rounded papilla 12. Development of juxta 13. Development of lateral excavations of vinculum 14. Lobes of uncus partially reduced 15. Development of very large cornuti in phallus* 16. Specific specialization of the shape of gnathos

54 17. Trophic specialization: Betulaceae (Betula, Carpinus, Alnus) and Po- aceae (Oplismenus) 18. Apical sclerotization of valva turned into an angular sclerite* 19. Lobes of uncus wide (mainly at basal parts) 20. Total reduction of cornuti (phallus without cornuti)* 21. Gnathos with narrow and long processes, and central plate turned into a transverse bar 22. Valva clearly triangle 23. Development of specialized paired carinae of phallus* 24. Secondary enlargement of ventral plate of vinculum and total or partial reduction of lobes 25. Trophic specialization: Carpinus 26. East Asiatic distribution (distribution range limited to East Asia) 27. Trophic specialization: Betula 28. Development of caudal tooth of the transverse bar of gnathos 29. Trans-Palaearctic distribution with the dominance of European distribution 30. Particular narrowing of secondary vinculum plate 31. Distinct lengthening of anterior processes of gnathos 32. Distinct development of lateral lobes of vinculum 33. Partial reduction of anterior excavation of vinculum 34. Reduction of anterior processes of gnathos 35. Appearance of angularity of transtilla 36. Development of lateral processes of transtilla 37. Trans-Palaearctic distribution with dominance of European distribution 38. Exceptional East Asian distribution 39. Increase in serration of caudal edge of tegumen 40. Narrowed fascia of forewing 41. Special arrangement of frass in a leaf-mine 42. Specific change in the shape of transtilla (total rounding of lateral angle)

* – Principal apomorphic characters supporting the clade (clades).

55 Fig. 50. Cladistic analysis of the Stigmella betulicola and the S. cornuta species groups of East Asia.

56 CONCLUSIONS

1. Morphological analysis of the main Nepticulidae material collected in East Asia and additional (comparative) material collected in the Ukraine, India, Mexico, and the countries of Central Asia and tropical Africa showed that the shape of valva, transtilla, vinculum, the development of uncus lobes, the number and shape of gnathos processes are also essential diagnostic characters of Nepticulidae species. The stability of these characters was registered after the analysis and comparison of material from very different and remote regions of the Earth; intra-specific or other variability of the above-mentioned structures that might affect the reliable species diagnostics was not found. 2. After re-examining the type series of Nepticulidae species described in the fauna of continental East Asia, it was found that descriptions of taxa prepared by other researchers from temporary micro-slides (in glycerine) of genital structures are not sufficiently precise and, in some cases, even confusing. An analysis of permanent micro-slides (in Euparol) prepared and digitally documented in this work for the first time allows stating that all examined East Asiatic Nepticuli- dae species have at least 1–3 additional characters of valvae, uncus, gnathos, phallus (including cornuti) or other sclerites that were not mentioned or correctly interpreted in the original descriptions of the species; though these characters are highly significant for diagnostics of taxa. 3. After analysis of unidentified scientific collection material of Neptic- ulidae collected by other researchers in continental East Asia, it was found that some earlier described species have to be merged (Stigmel- la regina was synonymized with S. sashai and Ectoedemia laura with E. sivickisi) because no diagnostic characters allowing these species to be regarded as independent taxa had not been detected, whereas the taxonomic status of S. auricularia, which is closely related to S. palionisi, was additionally supported by reliable characters of valvae and uncus. Some variability of the genital characters (though insignificant from the viewpoint of taxonomy) was noticed in a few East Asiatic species (e.g. Stigmella naturnella, Ectoedemia sivickisi);

57 however, morphological characteristics of most species were considerably supplemented not because of the noticed variability, but for the reason of insufficient species documentation by other researchers (including insufficient original descriptions): Stigmella sakhalinella, S. attenuata, S. alisa, S. dentatae, S. aladina, Ectoedemia chasanella, E. scoblei, E. olvina, E. ornatella, E. picturata, and many others. 4. The examination of the new material collected during the fieldwork revealed that diagnostics of Stigmella gimmonella, S. tranocrossa, and Ectoedemia philipi were based on inaccurate morphological characters, and E. philipi had a subbasal lobe of valva that had not been earlier known or described. The data on geographical distribution of some species (Ectoedemia ornatella, Etainia capesella, and others) were also updated. Stigmella betulicola and S. cathepostis, which earlier had been known only in the fauna of Japan, our study presented as distributed in continental East Asia; and S. thuringiaca, which earlier was known as a species exclusively with European distribution, has been for the first time detected in East Asia. 5. Six species which were described by us as new to science (Stigmella multispicata, S. sexcornuta, Ectoedemia ortiva, E. paraortiva, E. species 219 (in the fauna of Russian Far East), and S. cornuta (in the fauna of China)) differ from the earlier known species in the cornuti and other morphological characters of the male genitalia as well as in the arrangement of hindwing androconia (Ectoedemia ortiva, E. paraortiva). A total of 110 Nepticulidae species belonging to 5 genera: Stig- mella (69 species), Bohemannia (6), Ectoedemia (27), Fomoria (3), and Etainia (5), were discovered in East Asia. 6. Chorological analysis of the fauna showed that species detected in East Asia belong to 4 chorological groups. Most of them (91 species) belong to the East-Palaearctic group, yet on the basis of a more detailed analysis of distribution ranges this group can be split into three subgroups, the largest of which is the continental East-Palaearctic subgroup (43 species). The smallest chorological groups in East Asia include species with the broadest distribution ranges (the Holarc- tic and Amphi-Palaearctic). A comparison of chorological analysis with trophic relationships of species revealed that species with East-Palaearctic distribution are mostly trophically associated with

58 plants of Quercus, Acer, Malus, and Ulmus, whereas species with Trans-Palaearctic distribution are chiefly mining the Betula, Salix, and Populus plants. 7. On the basis of the new material from East Asia (i.e. data on biology and morphology of our newly described Nepticulidae species from northeastern China), we designated the cornuta group, a new species group of the Stigmella genus; a cladistic analysis showed that though this group has common characters with another Stigmella group (the betulicola), the new group is a separate (independent) evolutionary clade supported by at least 9 autapomorphic characters (the specific shape and very large size of cornuti, specific shape of gnathos with partly reduced caudal processes, unique leaf-mines with the final part widening into a large blotch; the combined leaf-mines are unique among all other species of the Stigmella genus trophically associated with the Fagaceae plants). In total, the species of Stigmella from East Asia can be assigned to 18 different species groups.

59 SANTRAUKA LIETUVIŲ KALBA

Disertacijos struktūra ir apimtis. Disertacija parašyta lietuvių kalba, disertacijos santrauka – anglų kalba. Disertaciją sudaro šie sky- riai: Įvadas, Literatūros apžvalga, Tyrimų metodai ir medžiaga, Tyrimų rezultatai, Išvados ir Literatūra (sudarytas 328 cituotų literatūros šaltinių sąrašas). Darbo apimtis – 211 puslapių, 379 paveikslai.

ĮVADAS

Problema ir tyrimų aktualumas. Mažieji gaubtagalviai (Nepticu- lidae) yra augalų asimiliacinius audinius minuojantys vabzdžiai, o kai kuriais atvejais – miško ir kultūrinių augalų kenkėjai (arba potencialūs kenkėjai) (Diškus & Stonis, 2012). Tačiau ne tik šiuo požiūriu mažieji gaubtagalviai yra svarbūs. Reikėtų atsižvelgti į tarptautinę Rio de Žanei- ro konvencija, kuri įgalino vykdyti ir plėtoti įvairius biologinės įvairovės tyrimus, siekiant inventorizuoti įvairaus rango taksonus (Stonis, 2010, 2011). Pastaruoju metu, tiek dėl klimato kaitos, tiek, svarbiausia, dėl žmo- gaus veiklos ir gyvūnų bei augalų gyvenamųjų vietų naikinimo bei fra- gmentacijos biologinė įvairovė sparčiai nyksta (Puplesis, 2002; Stonis, 2010). Šie procesai, menkai pastebimi mūsų šalyje, tačiau kituose kraš- tuose jau yra įgavę dramatišką pobūdį. Mūsų tyrimų objektas – filogenetiškai vienas primityviausių (ir tuo, teoriniu požiūriu, vienas svarbiausių bei įdomiausių) Lepidoptera būrio taksonų. Tiriama šeima, išsiskirianti ne tik archaiška sandara, bet ir labai didele specializacija, yra labai plačiai paplitusi visuose Žemės regionuose ir biotose (išskyrus Antarktį) (Puplesis, 1994; Diškus & Stonis, 2012). Nors Nepticulidae tyrimai pradėti vykdyti labai seniai, iki šiol dar daugelio regionų fauna yra menkai žinoma. Rytų Azija yra svarbus biologinės įvairovės centras, istoriškai ir faunos požiūriu artimai susijęs su Europos biota. Todėl Rytų Azijos Nepticulidae tyrimų rezultatai turi svarbią reikš- mę kalbant apie Eurazijoje vykusius faunogenetinius procesus. Maža to, dėl stenofagijos tendencijų, sėslaus gyvenimo būdo ir endemizmo mažieji

60 gaubtagalviai yra vieni puikiausių objektų, charakterizuojančių bet kurio tiriamo biomo (ar biotos) biologinės įvairovės turtingumą, kilmę ir gam- tinius ryšius (Stonis, 2010). Deja, šių, praktinę ir svarbią teorinę reikšmę turinčių vabzdžių tyrimų duomenų gerokai trūksta. Iki disertacijos metu atliktų tyrimų kontinentinėje Rytų Azijoje, mažųjų gaubtagalvių fauna buvo nepakankamai ištirta (Rocienė & Sto- nis, 2013; Stonis & Rocienė, 2013). Anksčiau paskelbti duomenys apie šio krašto Nepticulidae fauną buvo paremti neišsamiais drugių genitalinių struktūrų aprašais, o tipiniai individai ištirti ir dokumentuoti nepakankamai detaliai, remiantis tik laikinaisiais genitalijų mikropreparatais (Rocienė & Stonis, 2013). Laikinieji preparatai glicerine, skirtingai nuo pastoviųjų preparatų euparolyje, mažiau tinkami siekiant nuodugniai dokumentuoti persidengiančius skleritus ir lyginant morfologines struk- tūras tarpusavyje. Todėl tyrimams naudojant pastoviuosius preparatus, disertacinio darbo metu buvo išaiškinta ne tik naujų mokslui rūšių, bet ir žymiai patikslinti daugelio rūšių arealai, atlikta pirmoji Rytų Azijos Nepticulidae chorologinė analizė. Disertacinio darbo tikslas – ištyrus iki šiol neidentifikuotą moksli- nę kolekcinę medžiagą ir atlikus lauko darbus kontinentinėje Rytų Azi- joje, atlikti Rytų Azijos Nepticulidae faunos taksonominę ir chorologinę analizę. Darbo uždaviniai: 1. Remiantis nauja Nepticulidae medžiaga, surinkta Rytų Azijoje ir nauja papildoma (palyginamąja) medžiaga, surinkta Ukrainoje, Indijo- je, Meksikoje ir Centrinės Azijos bei atogrąžų Afrikos šalyse, nustatyti ir įvardyti šeimos morfologinius požymius, kurie šiuolaikinių tyrimų kontekste gali ir turėtų būti panaudoti šeimos taksonų (ypač rūšių) diagnostikoje. 2. Parengus pastoviuosius (euparolinius) mikropreparatus ir naujuo- ju metodu ištyrus bei dokumentavus kontinentinės Rytų Azijos faunoje aprašytų Nepticulidae rūšių tipines serijas, nustatyti, ar visų rūšių pir- miniai aprašai yra visiškai tikslūs ir patikimai charakterizuoja analizuo- jamas rūšis (įvardijant ir aprašant konkrečius genitalinių struktūrų po- žymius, jeigu požymiai bus nustatyti kaip iki šiol netiksliai interpretuoti arba neaprašyti).

61 3. Ištyrus iki šiol neidentifikuotą kitų tyrėjų kontinentinėje Rytų Azijoje surinktą mokslinę kolekcinę Nepticulidae medžiagą, atlikus taksonominius keitimus ir aptikus bei įvertinus galimus požymių kintamumo atvejus, patikslinti rūšių morfologines charakteristikas. 4. Atlikus ekspedicinius lauko darbus ir ištyrus surinktą naują bei kitų tyrėjų medžiagą, patikslinti rūšių diagnostinius požymius ir geografinio paplitimo duomenis. 5. Išaiškinti ir aprašyti naujas mokslui rūšis ir įvertinti bendrąją Rytų Azijos Nepticulidae taksonominę įvairovę. 6. Atlikti Rytų Azijos Nepticulidae chorologinę analizę ir nustatyti kaip chorologinės analizės rezultatai siejasi su rūšių trofiniais ryšiais. 7. Remiantis nauja Rytų Azijos medžiaga, išskirti ir aprašyti naują rūšių grupę Stigmella gentyje, integruojant kladistinės analizės duomenis ir pateikiant papildomų įrodymų apie naujos grupės išskirtinumą. Mokslinis darbo naujumas. Atrasta, aprašyta ar įvardinta ir parengta aprašymui 14 naujų mokslui mažųjų gaubtagalvių rūšių iš Rytų Azijos (Primorės kraštas), Kinijos ir Indijos. Dviejų Rytų Azijos Stigmella rūšių ir dviejų Ectoedemia rūšių pavadinimai sinonimizuoti. Pirmą kartą ištyrinėta, preparuota ir aprašyta iki šiol neidentifikuota Nepticulidae medžiaga, saugoma Rusijos MA Zoologijos institute. Pirmą kartą ištirtos visų Rusijos Tolimuosiuose Rytuose ankščiau aptiktų ir aprašytų 56 Nepticulidae rūšių tipinės serijos, padaryti visų rūšių holotipų pastovieji morfologiniai mikropreparatai, o rūšių aprašai papildyti naujais duomenimis ir dokumentuoti pagal šiuolaikinius moks- linius standartus. Pirmą kartą atliktas Rytų Azijos Nepticulidae faunos chorologinis įvertinimas, naujais rūšių duomenimis papildytas faunos taksonominis sąvadas, išanalizuoti trofiniai ryšiai. Atliekant šiuos tyrimus buvo parengtas papildytas Rytų Azijos Nep- ticulidae sistematinis sąrašas (iš viso 110 rūšių), pirmą kartą apibendrinti Rytų Azijos faunos rūšių trofinių ryšių ir geografinio paplitimo ypatu- mai. Tiriant kitų kraštų fauną (t. y. atliekant palyginamuosius tyrimus), pirmą kartą sudarytas Meksikos (Centrinė Amerika) mažųjų gaubtagal- vių taksonominis sąvadas. Naujai revizuota Centrinės Azijos Acalyptris fauna, atskleidžiant rūšių taksonomijai reikšmingus morfologijos ypatu-

62 mus, pirmą kartą preparuota gausi kolekcinė medžiaga, surinkta atogrą- žų Afrikoje. Mokslinė ir praktinė reikšmė. Pastaruoju metu, siekiant vystyti įvairius biologinės įvairovės tyrimus, skatinama vis intensyviau inventorizuoti pasaulio biologinę įvairovę, atlikti taksonominius tyrimus, aprašyti iki šiol nežinomas mokslui rūšis (Stonis, 2010, 2011). Šiuo metu aktyviai pradėti tirti ir endobiontiniai vabzdžiai, kurių lervos maitinasi ir vystosi žaliuosiuose (asimiliaciniuose) augalo audiniuose (Diškus & Stonis, 2012). Toks gyvenimo būdas dar yra vadinamas minavimu, o augalų asimiliaciniuose audiniuose padaryti pažeidimai vadinami mi- nomis. Šioje disertacijoje nagrinėjama mažųjų gaubtagalvių (Nepticulidae) šeima, kuri nors ir plačiai paplitusi visuose sausumos ekosistemose, ta- čiau tiek praktiniu, tiek ir teoriniu aspektu, vis dar nepakankamai išty- rinėta. Daugelis mažųjų gaubtagalvių rūšių dėl jų vikšrų endobiontinio gyvenimo būdo augalų audiniuose yra svarbūs ūkiniu požiūriu, nes dalis šių organizmų gali būti pripažinti kaip potencialūs kenkėjai (Kuznetzov & Puplesis, 1994; Puplesis, 1994). Atlikti minuojančių vabzdžių faunos tyrimai ir surinkti duomenys apie jų mitybinę specializaciją ir paplitimą, gali būti panaudoti augalų apsaugai, kuriant įvairias augalų apsaugos priemones (Diškus & Stonis, 2012). Kita vertus, mažieji gaubtagalviai (kaip ir kai kurie endobiontiniai organizmai) yra gana sėslūs nors aptinkami beveik kiekvienoje sausumos ekosistemoje. Jų tyrimai leidžia geriau suvokti biologinės įvairovės sudėtį, evoliucines tendencijas bei biogeografines ypatybes (Stonis, 2010). Taksonomiškai inventorizuodami Rytų Azijos fauną, gavome duomenų (Rocienė & Stonis, 2013; Stonis & Rocienė, 2013, 2014), kurie keičia iki šiol nusistovėjusią sampratą apie netolygų Nepticulidae pasiskirstymą įvairiuose Žemės regionuose, galimą Rytų Azijos faunos kilmę ir zoogeo- grafinius ryšius su giminiška Euro-Nemoraline fauna. Ginamieji darbo teiginiai: 1. Nepticulidae morfologijos požymiai yra svarbūs šiuolaikinių ty- rimų kontekste, tačiau įvairių požymių reikšmė yra ne vienoda; patinų genitalinių struktūrų požymiams yra būdingas rūšinis specifiškumas ir pastovumas, nesvarbu kurio Žemės regiono fauna būtų nagrinėjama.

63 2. Rytų Azijos mažųjų gaubtagalvių rūšims yra būdingi įvairūs, anksčiau nedokumentuoti (arba klaidingai interpretuoti) genitalinių struktūrų požymiai, turintys svarbią reikšmę taksonų diagnostikoje. 3. Kai kurios ankščiau aprašytos Rytų Azijos rūšys yra neparemtos jokiais požymiais, kurie leistų šias rūšis traktuoti kaip savarankiškus taksonus, todėl turi būti apjungtos, o kitų Rytų Azijos rūšių morfologi- nės charakteristikos papildytos. 4. Kai kurių Rytų Azijos rūšių diagnozės iki šiol rėmėsi netiksliais morfologiniais požymiais, o dalies rūšių geografinis paplitimas gali būti papildytas esminiais naujais duomenimis. 5. Rusijos Tolimuosiuose Rytuose aptinkamos Stigmella multispicata, S. sexcornuta, Ectoedemia ortiva, E. paraortiva, E. species 219 ir Kinijoje aptinkama Stigmella cornuta yra naujos mokslui rūšys, kurios skiriasi nuo ankščiau žinomų rūšių įvairiais, aiškiai išreikštais požymiais. Iš viso Rytų Azijoje šiuo metu yra žinoma 110 mažųjų gaubtagalvių rūšių, kurių dauguma priklauso Stigmella genčiai ir tik nedaugelis rūšių kitoms gentims. 6. Chorologiniu požiūriu Rytų Azijos Nepticulidae fauna nėra viena- lytė; ją sudaro 4 skirtingų chorologinių grupių rūšys; tarp jų vyraujantys yra rytų palearktinio arealo taksonai, kurie trofiškai susiję su Quercus, Acer, Malus ir Ulmus, o plačiausių arealų Nepticulidae rūšys sudaro tik nežymią mažumą. 7. Stigmella cornuta yra nauja Stigmella genties rūšių grupė, kuri nors ir turi bendrų požymių su Stigmella betulicola grupe, tačiau išsi- skiria dėl specifinės cornuti formos ir dydžio, specifinės gnathos formos, iš dalies redukuotų kaudalinių išaugų ir kombinuotos minos mitybinio augalo lape. Tyrimų rezultatų publikavimas. Disertacijoje nagrinėjamų tyrimų rezultatai paskelbti 12 darbų, kurie išspausdinti moksliniuose žurna- luose, tęstiniuose ir vienkartiniuose leidiniuose. Iš jų penki moksliniai straipsniai paskelbti Zootaxa žurnale (su citavimo indeksu impact factor), du – straipsniai paskelbti periodiniuose ISI sąrašo (Master Journal List) mokslo žurnaluose Acta Zoologica Lituanica ir Ecology and Zoology, vienas straipsnis ir vienos tezės – tarptautinių mokslinių konferencijų me- džiagoje, du straipsniai – kituose leidiniuose. Kartu su bendraautoriais taip pat publikuotas vienas monografijos (Diškus & Stonis, 2012: Lietu-

64 vos Nepticulidae faunos taksonominė, chorologinė ir trofinė charakteristika) skyrius ir vienas metodinis leidinys. Iš viso autorė kartu su bendraautoriais paskelbė: 10 darbų anglų kalba ir 2 darbus lietuvių kalba. Padėkos. Nuoširdžiai dėkoju savo mokslinio darbo vadovui prof. habil. dr. Jonui Rimantui Stoniui (LEU) už pasiūlymą gilintis į šią įdo- mią temą, pagalbą gaunant tyrimų medžiagą bei įvairiapuses pastabas ruošiant šį darbą; darbo moksliniam konsultantui prof. dr. Virginijui Sruogai (LEU) dėkoju už vertingus patarimus ir padrąsinimą. Už įdo- mias diskusijas dėkoju Lietuvos edukologijos universiteto Biologijos katedros kolegoms ir ypač Biosistematikos tyrimų grupės nariams doc. dr. A. Diškui, A. Remeikiui ir A. Navickaitei. Už svetingą priė- mimą, malonų ir draugišką bendradarbiavimą bei vertingus patarimus lankantis Rusijos MA Zoologijos instituto (Sankt Peterburgas) Vabz- džių sistematikos laboratorijoje, esu dėkinga laboratorijos vedėjui dr. S. Yu. Siniovui (С. Ю. Синёв), bei kitiems kolegoms – dr. A. L. Lvovskiui (А. Л. Львовский), dr. S. V. Baryšnikovai (С. В. Барышникова) ir dr. V. G. Mironovui (В. Г. Миронов). Taip pat dėkoju Tervureno (Belgija) Karališkojo Centrinės Afrikos muziejaus Lepidoptera kolekcijos kura- torei dr. J. De Prins už suteiktą kolekcinę medžiagą tyrimams. Nuošir- džiai dėkoju Rusijos MA Tolimųjų Rytų skyriaus (Primorės kraštas) mokslinės stoties „Gornotajožnaja“ direktoriui prof. dr. P. S. Zorikovui (П. С. Зориков) už suteiktą galimybę vykdyti tyrimus bei techninę įran- gą lauko ir laboratorinių tyrimų atlikimui. Taip pat nuoširdžiai dėkoju šios stoties Vabzdžių ekologijos laboratorijos vedėjui dr. M. M. Omelko (M. M. Омелько) ir jo žmonai dr. N. V. Omelko (Н. В. Омелько), už svetingą ir nuoširdų priėmimą bei visokeriopą pagalbą. Už svarbius ir draugiškus patarimus ir nuoširdžias diskusijas esu labai dėkinga prof. dr. J. Trimblui (JAV) ir dr. Andriui Petrašiūnui (VU). Dėkoju savo draugams ir ypač šeimai – vyrui Donatui, mamai Ange- lei, tėčiui Romui, krikšto tėvams Romai ir Valdui, už begalinę kantrybę, toleranciją, supratingumą bei palaikymą. Doktorantūros studijos, šiuo laikotarpiu atlikti ekspediciniai lauko ir laboratoriniai tyrimai buvo finansuojami iš Europos Sąjungos struk- tūrinių fondų projekto „Aukštos kvalifikacijos specialistų atitinkančių valstybės ir visuomenės poreikius biomedicinos srityje rengimo tobulini-

65 mas – BIOMEDOKT“, o taip pat ir Valstybinio studijų fondo bei Lietuvos mokslų tarybos lėšomis.

LITERATŪROS APŽVALGA

Išsami literatūros apžvalga yra pateikta disertacijoje ir užima 14 spausdintų puslapių. Joje yra pateikiamos: bendroji Nepticulidae litera- tūros apžvalga, pastarojo laikotarpio svarbiausių publikacijų apžvalga bei Rytų Azijos Nepticulidae tyrimų apžvalga.

TYRYMŲ METODAI IR MEDŽIAGA

Tyrimų metodai. Vienas efektyviausių mažųjų gaubtagalvių rinkimo būdų yra jų viliojimas šviesa. Vabzdžiams gaudyti buvo nau- dojamas baltas apie 2 × 3 m dydžio audinio ekranas, kurio viršutinis kraštas užlenkiamas apie 1 cm ir prisiuvamas taip, kad į susidariusį plyšį būtų galima įverti nestorą, bet tvirtą virvę. Virvės ilgis paprastai buvo 20–30 metrų, tam, kad būtų patogu ištemti ekraną tarp medžių. Prie ekrano viršutinio krašto (centre), aukščiau akių lygio, trišakiu laikikliu buvo pritvirtinama lempa. Tinkamiausios Nepticulidae vilioti yra DRL ar LRF tipo ultravioletinių spindulių lempos arba jų analogai. Ekranas įtempiamas, prispaudžiant ir užlenkiant į priekį apatinį jo kraštą. Apati- nis ekrano kraštas ant žemės užlenkiamas tam, kad galima būtų surinkti nukritusius drugius. Ekranas buvo įtempiamas taip, kad jo nesiūbuotų vėjas (vėjuotų vietų buvo vengiama). Nuo įtempto ekrano lengviau nu- imti drugius, o blogai įtempus lieka raukšlės, kuriose sunku pastebėti ir sugauti drugį. Nuo ekrano mažieji gaubtagalviai buvo nuimami mažais (maždaug 5–8 mm skersmens ir 3–5 cm ilgio) stikliniais mėgintuvėliais, užkimštais nedideliu vatos gumulėliu. Kad drugiai kuo mažiau nusi- trintų žvynelius, kiekvienas drugys buvo dedamas į atskirą mėgintuvėlį. Mėgintuvėlio kamštis turi būti pakankamai standus, kad neiškristų, bet kartu ir pakankamai pralaidus, kad pro jį praeitų marinimo medžiagos

66 (etilacetatas). Mėgintuvėlis su drugiu iškarto dedamas į nuodytuvą – sandariai ir lengvai uždaromas indas (paprastai stiklinis), kurio dugne įdėtas 3–5 cm aukščio vatos įklotas. Be šviesinių gaudyklių, suaugę drugiai buvo renkami nedideliu (15 cm skersmens) entomologiniu tinkleliu braukiant virš mitybinių mažųjų gaubtagalvių augalų. Kad vabzdžiai kuo mažiau apsitrintų, surinkta medžiaga buvo tvarkoma iš karto, arba ne vėliau kaip kitos dienos pirmoje pusėje. Prieš tai medžiaga buvo laikoma tamsioje ir vėsioje vietoje (pvz., šaldytuve). Mūsų darbe taip pat buvo taikomas svarbus, daug laiko ir darbo reikalaujantis Nepticulidae medžiagos rinkimo būdas – suaugėlių augi- nimas iš minuojančių vikšrų. Naudojant šį metodą metu ne tik gaunama aukščiausios kokybės medžiaga, morfologiškai kokybiški individai, bet ir itin svarbūs duomenys apie mitybinius augalus, minos tipą, kokono spal- vą ir dydį bei vystymosi ciklą. Genitalinės struktūros (genitalinis aparatas) yra ypač svarbus mor- fologinių požymių kompleksas Nepticulidae diagnostikai ir sistemati- kai. Be šių struktūrų tyrimų bei panaudojimo neįmanoma patikimai identifikuoti ar aprašyti naujų rūšių. Nepticulidae genitaliniai požymiai (kaip ir kitų Lepidoptera) yra mažai kintantys ir specifiški rūšies ar net aukštesnės sistematinės grupės lygmeniu (Šimkevičiūtė [Rocienė] et al., 2010). Tyrimų metu buvo daromi laikinieji ir pastovieji mikropreparatai. Laikiniesiems ir pastoviesiems mikropreparatams stebėti ir morfologijai nagrinėti buvo naudojami tik didelio didinimo tiriamieji mikroskopai. Genitalinės struktūros buvo matuojamos ir fotografuojamos, o skleritai aprašomi. Visi padaryti preparatai buvo registruojami mikropreparatų žurnale, nurodant padaryto preparato numerį, rūšies pavadinimą, prepa- ravimo padarymo datą, instituciją, kuriai priklauso preparuotas indivi- das ir įvairios kitos pastabos apie konkretų preparatą. Rūšių diagnostikai svarbių genitalinių struktūrų ilgiai buvo matuojami tiek laikinuosiuose (t. y. glicerine), tiek pastoviuosiuose preparatuose (fiksuotose euparalyje), naudojant stereoskopinį mikroskopą Leica DM 2500 ir skaitmeninę kamerą Leica DFC 420 bei programinę įrangą Leica Application Suite V3.0.0. Suaugėliai buvo matuojami naudojant stereosko- pinį mikroskopą MBS-10 su okuliarine matavimo liniuote, o jų išorė fo- tografuojama naudojant stereoskopinį mikroskopą Leica S6D, fotografa- vimo kamerą DFC 290 ir programinę įrangą Leica Image Manager IM50.

67 Tyrimų medžiaga. Rengiant disertaciją mokslinė Nepticulidae medžiaga buvo renkama 2010–2013 metais ekspedicinių tyrimų metu Indijoje (Himalajuose, 2010 m.), Rusijos MA Tolimųjų Rytų skyriaus biologinių tyrimų stotyje „ Gornotayezhnaya“ (Usurijsko raj., 2011 m.) ir šiaurės rytinėje Kinijoje (2013 m.); papildomos mažųjų gaubtagalvių paieškos buvo atliktos lauko darbų metu šiaurės Tailande. Moksliniams tyrimams Nepticulidae medžiaga laikinai buvo pasiskolinta iš Rusijos MA Zoologijos instituto (Sankt Peterburgas). Taip pat buvo preparuota ir ištirta (Šimkevičiūtė [Rocienė] et al., 2009) medžiaga, surinkta kitų tyrėjų Meksikoje, Ukrainoje, Centrinėje Azijoje ir atogrąžų Afrikoje. Iš viso rengiant disertaciją buvo ištirta apie 3500 Nepticulidae individų. Paruošti visų minėtų individų laikinieji ir daugiau kaip 600 pastoviųjų genitalinių struktūrų mikropreparatų.

68 TYRIMŲ REZULTATAI (IŠVADOS)

1. Morfologiniai pagrindinės Nepticulidae medžiagos, surinktos Rytų Azijoje ir papildomos (palyginamosios) medžiagos, surinktos Ukrai- noje, Indijoje, Meksikoje ir Centrinės Azijos bei atogrąžų Afrikos šalyse, tyrimai parodė, kad uncus skiautėtumas, gnathos išaugų skai- čius bei jų forma yra esminiai Nepticulidae rūšių diagnostikos požy- miai. Registruotas šių požymių pastovumas analizuojant ir lyginant medžiagą iš labai skirtingų bei nutolusių Žemės regionų; aukščiau išvardytų esminių struktūrų vidurūšinis ar kitoks kintamumas, ga- lintis turėti įtakos patikimai rūšių diagnostikai, nenustatytas. 2. Ištyrus kontinentinės Rytų Azijos faunoje aprašytų Nepticulidae rū- šių tipines serijas, nustatyta, kad aprašytų taksonų aprašai, kitų tyrė- jų parengti vien tik remiantis laikinaisiais (glicerininiais) genitalinių struktūrų mikropreparatais, nėra pakankamai tikslūs, o atskirais atvejais – klaidinantys. Darbo metu pirmą kartą paruoštų ir skaitme- niniu būdu dokumentuotų pastoviųjų (euparolinių) mikropreparatų analizė leidžia teigti, kad visoms ištirtoms Rytų Azijos mažųjų gaub- tagalvių rūšims yra būdingi 1–3 papildomi valvae, uncus, gnathos, phallus (įskaitant cornuti) ar kitų skleritų požymiai, kurie nebuvo paminėti arba teisingai interpretuoti pirminiuose rūšių aprašuose; šie ankščiau neįvertinti požymiai turi svarbią reikšmę taksonų diagnostikoje. 3. Ištyrus iki šiol neidentifikuotą mokslinę kolekcinę Nepticulidae me- džiagą, kitų tyrėjų surinktą kontinentinėje Azijoje, nustatyta, kad kai kurios ankščiau aprašytos rūšys (Stigmella regina ir S. sashai bei Ectoedemia laura ir E. sivickisi) turi būti apjungtos, nes neaptikta diagnostinių požymių, kurie leistų šias rūšis traktuoti kaip savaran- kiškus taksonus. Tuo tarpu S. palionisi rūšiai giminiškos S. auricularia taksonominis statusas papildomai paremtas patikimais valvae ir uncus morfologijos požymiais. Kai kuriose kontinentinės Rytų Azijos rūšyse pastebėtas taksonominiu požiūriu nereikšmingas genitalinių požymių kintamumas (Stigmella naturnella, Ectoedemia sivickisi), tačiau daugelio rūšių morfologinės charakteristikos ženkliai papildytos ne dėl pastebėto kintamumo, o dėl ne pakankamos, kitų tyrėjų ankščiau atliktos rūšių dokumentacijos (įskaitant nepakankamus

69 pirminius aprašus): Stigmella sakhalinella, S. attenuata, S. alisa, S. dentatae, S. aladina, Ectoedemia chasanella, E. scoblei, E. olvina, E. ornatella, E. picturata ir daugelis kitų. 4. Ištyrus ekspedicinių lauko darbų metu surinktą naują medžiagą, nustatyta, kad Stigmella gimmonella, S. tranocrossa ir Ectoedemia philipi diagnozės rėmėsi netiksliais morfologiniais požymiais; E. philipi yra būdinga iki šiol buvusi nežinoma ir neaprašyta porinės plokštelės (valva) priešpamatinė skiautė. Taip pat patikslinti įvairių rūšių geografinio paplitimo nuomenys Ectoedemia( ornatella, Etainia capesella ir kt.). Stigmella betulicola ir S. cathepostis, kurios ankščiau buvo žinomos tik Japonijos faunoje, dabar pateiktos kaip paplitusios ir kontinentinėje Rytų Azijoje; nustatyta, kad S. thuringiaca, kuri ankščiau buvo žinoma kaip išimtinai europinio paplitimo rūšis, taip pat yra aptinkama Rytų Azijoje. 5. Tyrimu metu aprašytos 6 naujos mokslui rūšys (Stigmella multispicata, S. sexcornuta, Ectoedemia ortiva, E. paraortiva, E. species 219 Ru- sijos Tolimųjų Rytų faunoje ir S. cornuta Kinijos faunoje) skiriasi nuo ankščiau žinomų rūšių cornuti ir kitų patino genitalijų morfologijos požymiais, o taip pat užpakalinio sparno androkonijų išsidėstymu (Ectoedemia ortiva, E. paraortiva). Iš viso Rytų Azijoje išaiškinta 110 mažųjų gaubtagalvių rūšių, priklausančių penkioms gentims: Stigmella (69 rūšys), Bohemannia (6 rūšys), Ectoedemia (27 rūšys), Fomoria (3 rūšys) ir Etainia (5 rūšys). 6. Chorologinė faunos analizė parodė, kad Rytų Azijoje aptinkamos rū- šys priklauso 4 skirtingoms chorologinėms grupėms. Daugiausiai rū- šių (91 rūšis) priklauso rytų palearktinei grupei, tačiau remiantis de- talesne arealų analize, ši grupė gali būti suskirstyta į tris pogrupius, iš kurių didžiausias yra rytų palearktinis-kontinentinis (43 rūšys). Mažiausios Rytų Azijoje yra tos chorologinės grupės, kurių rūšims būdingi patys plačiausi arealai (holarktinė ir amfipalearktinė choro- loginės grupės). Palyginus chorologinės analizės duomenis su rūšių trofiniais ryšiais, nustatyta, kad rytų palearktinio paplitimo grupei daugiausiai būdingas Quercus, Acer, Malus ir Ulmus minavimas, o tuo tarpu transpalearktinio paplitimo rūšims daugiausiai būdingas Betula, Salix ir Populus minavimas.

70 7. Remiantis nauja Rytų Azijos medžiaga (t.y. duomenimis apie mūsų aprašytos, šiaurės rytų Kinijoje paplitusios naujos mažųjų gaubta- galvių rūšies biologiją ir morfologiją), išskirta nauja Stigmella genties rūšių grupė – cornuta; kladistinė analizė parodė, kad nors ši grupė turi bendrų požymių su kita Stigmella genties rūšių grupe (betulicola), nauja rūšių grupė yra atskirta (savarankiška) evoliucinė šaka, kuriai būdingi mažų mažiausiai 9 autapomorfiniai požymiai (specifinė cornuti forma ir neįprastai didelis jų dydis, specifinėgnathos forma su iš dalies redukuotomis kaudalinėmis išaugomis, sudėtingos vikšrų išgraužtos minos, kurių galinė dalis išplatėja į didelę dėmę; tokios kombinuotos minos yra unikalios tarp visų kitų Stigmella genties rūšių, kurios trofiškai susijusios su Fagaceae šeimos augalais). Iš viso Rytų Azijoje aptinkamos Stigmella genties rūšys gali būti priskirtos 18-ai skirtingų rūšių grupių.

71 LIST OF PUBLICATIONS CONTAINING MATERIALS OF THE THESIS

CHAPTER OF MONOGRAPH

1. Stonis, J. R., Diškus, A., Navickaitė, A., Remeikis, A., Rocienė, A. 2012. Morfologiniai Nepticulidae požymiai, naudojami identifikuo- jant taksonus ir paremiantys šiuolaikinę šeimos sistemą. In: Diškus, A., Stonis, J. R. Lietuvos endobiontiniai vabzdžiai. Nepticulidae faunos taksonominė, chorologinė ir trofinė charakteristika. Monografija. Lututė, Kaunas. 60–72 p.

PAPERS IN SCIENTIFIC PERIODICAL JOURNALS

2. Šimkevičiūtė [Rocienė], A., Stonis, J. R., Diškus, A. 2009. Taxonomic checklist of Nepticulidae of Mexico, with the description of three new species from the Pacific Coast (Insecta, Lepidoptera). Acta Zoo- logica Lituanica, 19 (4): 268–277. 3. Stonis, J. R., Navickaitė, A., Rocienė, A., Remeikis, A., Diškus, A. 2013. A provisional checklist of the Nepticulidae (Insecta, Lepidopte- ra) of the Crimea. Zoology and Ecology, 23 (1): 1–9. 4. Stonis, J. R., Rocienė, A. 2013. Nepticulidae (Lepidoptera) of East Asia (1). Re-examination of the male genitalia of types deposited at the Russian Academy of Sciences. Zootaxa, 3652 (1): 1–59. 5. Rocienė, A., Stonis, J. R. 2013. Nepticulidae (Lepidoptera) of East Asia (2). Study of a collection sample deposited at the Russian Aca- demy of Sciences, with descriptions of new species and a checklist. Zootaxa, 3652 (2): 75–116. 6. Stonis, J. R., Diškus, A., Remeikis, A., Navickaitė, A., Rocienė, A. 2013. Description of new species of oak leaf-miners (Lepidoptera: Nepticulidae), with notes on the species groups of Stigmella Schrank associated with Quercus as a host-plant. Zootaxa, 3737 (3): 201–222. 7. Stonis, J. R., Diškus, A., Rocienė, A., Sruoga, V., Davis, D. R. 2014. New and little known Coptotriche and Tischeria species (Lepidoptera: Tischeriidae) from Primorskiy Kray, East Asia. Zootaxa [accepted].

72 8. Stonis, J. R., Rocienė, A. 2014. Additions to the Nepticulidae (Lepi- doptera) of East Asia, with descriptions of three new species from Primorskiy Kray. Zootaxa [accepted, in press].

PAPERS AND ABSTRACTS OF SCIENTIFIC CONFERENCES

9. Šimkevičiūtė [Rocienė], A., Stonis, J. R., Sruoga, V. 2010. Male genitalia characters and their diagnostic value in taxonomy of Nepticu- lidae (Insecta: Lepidoptera). Proceedings of 5th International Confe- rence “From Biotechnology to Environment Protection”, Zielona Gora, 25–27 November, 2010, University of Zielona Gora Publishers, p. 44. 10. Rocienė, A., Stonis, J. R. 2014. Revised fauna of the Nepticulidae (Lepidoptera) of continental East Asia: lots of effort to elucidate the little known diversity of pygmy moths. In: Stonis, J. R., Diškus, A., Valainis, U., Jackowski, J., Auškalnis, T. & Jurkonytė, A. (eds). Selec- ted abstracts and papers of the First Baltic International Conference on Field Entomology and Faunistics. Edukologija Publishers, Vilnius.

OTHER PUBLICATIONS

11. Noreika, R., Šimkevičiūtė [Rocienė], A. 2011. Drugių tyrimo me- todika. In: Stonkutė, R. (sud.). Jaunojo tyrėjo vadovas, t. d. Vilnius: Lietuvos mokinių informavimo ir techninės kūrybos centras, p. 113– 151. 12. Rocienė, A. 2011. LEU entomologijos doktorantų tyrimai siekia net Tolimuosius Rytus. Šviesa, 12 (2409): 2.

73 CURRICULUM VITAE

AGNĖ ROCIENĖ

Personal information: Lithuanian; born 13 August 1985, in Rad- viliškis, Lithuania. Education: 2004–2008, Bachelor’s Degree in Ecology, Fac- ulty of Natural Sciences, Vilnius Pedagogical University; 2008–2010, Master’s Degree in Biology, Faculty of Natural Sciences, Vilnius Pedagogical University; 2010–2014, Doctoral studies (PhD fellow), Lith- uanian University of Educational Sciences. Current appointment: Vice-Dean of the Faculty of Science and Tech- nology, Lithuanian University of Educational Sciences; Lecturer at the Department of Biology, Faculty of Science and Technology, Lithuanian Univer- sity of Educational Sciences. Project activities: National project financed by the Lithuanian State Studies and Research Foundation “Devel- opment of Ecosystems and Molecular Investi- gation of the Phylogeography of Populations in the Baltic Region”: “New Phylogenetic Hypoth- eses about Plant-Mining Primitive Lepidoptera Based on Molecular Data (Lepidoptera: Neptic- uloidea, Tischeriodea, Gelechiodea)”(2009); National project financed from the Research Foundation of the Research Council of Lithua- nia “New Faunas: Taxonomic Analysis and En- dobiontic Lepidoptera Species New to Science (MIP-049/2011)” (2011–2012). Research visits: Zoological Institute of the Russian Academy of Sciences, St. Petersburg, Russia (2010, 2011, 2012) Contacts: Room 216, Faculty of Science and Technology, Lithuanian University of Educational Sciences, Studentų g. 39, Vilnius, LT-08106 E-mail: agne.rociene @leu.lt

Agnė Rocienė

TAXONOMY OF THE NEPTICULIDAE OF CONTINENTAL EAST ASIA (INSECTA, LEPIDOPTERA, NEPTICULIDAE)

Summary of Doctoral Dissertation Biomedical Sciences, Biology (01B): Entomology and Plant Parasitology, B–250

Maketavo Laura Petrauskienė

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