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FIGURE 3 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 3. COI Neighbor-Joining tree of Gracillariidae species sampled in the Russian Far East. Each specimen is accompanied by its sequence number (BOLD "process ID"), species name, sampling location and host plant. Sampling locations are given between two vertical lines, i.e. |Skov.| Skovorodino; |Blag.| Blagoveshchensk; |MTS| Komarov Mountain-Taiga Station FEB RAS and the forest around; |Rakovka|, |Glukhovka| forest around the villages Rakovka and Glukhovka respectively; |Vlad.| Vladivostok; |Sikhote-Alin| Sikhote-Alin Mountains, national park "Zov tigra", |Y-S| Yuzhno-Sakhalinsk and the area around including Susunay mountain range (see Fig. 1 for details). Host plants: A. pseudosieb.—Acer pseudosieboldianum; * sampling done by sweep netting from the bush, whereas all other gracillariids were collected/ reared directly from their mines. Each leafminer species is supplied with its BIN number retrieved from BOLD; blue BINs correspond to known species, red—new BINs.

FIGURE 1 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 1. The area of study in the Russian Far East, June–July 2010–2017. Black circles indicate cities, yellow circles with the numbers 1–12 are the sampling locations: (1) AO, Skovorodino, tree plantation nearby train station; (2) AO, Blagoveshchensk, Friendship park; (3) KK, Komsomolsk-na-Amure, tree lines along the road; (4) PK, Gornotaezhnoe, MTS; (5) ibidem, forest; (6) PK, Glukhovka, forest; (7) PK, Rakovka, forest; (8) PK, Vladivostok, suburb, tree plantation; (9) PK, Sikhote-Alin Mountains, national park "Zov tigra"; (10) SO, Sakhalin Isl., Yuzhno-Sakhalinsk, Botanical garden FEB RAS Sakhalin Branch; (11) ibidem, Gagarin park; (12) SO, Sakhalin Isl., Susunay mountain range, pass "Verblyud". In the left corner, on the map of Russia the sampled area is shaded in gray. Regions: (ChAO) Chukotsky Autonomous Okrug, (KamK) Kamchatskii Krai, (MA) Magadanskaya Oblast, (AO) Amurskaya Oblast, (KK) Khabarovskii Krai, (JAO) Jewish Autonomous Oblast, (AO) Amurskaya Oblast, (PK) Primorskii Krai, (SO) Sakhalinskaya Oblast (includes Sakhalin Island and Kuril Islands).

FIGURE 7 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 7. Male genitalia of Phyllonorycter spp. from the Russian Far East. (A) Phyllonorycter junoniella, genitalia slide [21-male]; (B) Ph. pastorella, host plant Salix sp., [12Pp-2016-male]; (C) Ph. populifoliella, Populus balsamifera, [NK-561- 1-male]; (D) Ph. pseudojezoniella, Acer saccharum, [20Pps-2016-male]; (E) Ph. kisoensis, Alnus hirsuta, [5Pk-2016-male]. Sampling location: (A) SO, Sakhalin Isl., 14.VII.2017; (B, D) PK, Gornotaezhnoe, MTS, 22–26.VII.2016; (C) AO, Blagoveshchensk, 27.VI.2016. Scale bars 200µm.

FIGURE 11 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 11. Mines and leaf shelters of Gracillaria, Callisto, Parornix and Phyllonorycter spp. from the Russian Far East. (A) Gracillaria sp., host plant Syringa amurensis; (B–C) Callisto sp. on Malus sp.; (D) Parornix ermolaevi, Corylus sieboldiana; (E–F) Ph. caraganella, Caragana fruticosa; (G–H) Ph. cavella, Betula platyphylla, (H) "green island" and the mine. Indications: (m) mine; (b) blotch part of the mine; (t) epidermal tunnel; (e) exit hole; (f–f2) folded leaf margin or leaf tip (the indexes 1 and 2 indicate the order of construction appearance); (g.i.) green island region. Close up: (B, E–G) mine. Sampling locations: (A–D, G–H) PK, Gornotaezhnoe, MTS and forest, 22–27.VII.2016; (E–F) ibidem, Glukhovka, forest, 27.VII.2016.

FIGURE 2 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 2. Sampling localities in the Russian Far East. (A–C) PK, Gornotaezhnoe, MTS: (A) European woody plant species zone, (B) Juglans mandshurica plantation, (C) East Asian woody plant species zone; (D) PK, 20 km west of MTS, artificial lake on the way to Glukhovka and Rakovka; (E–H) SO, Sakhalin Isl., Susunay mountain range: (E–G) pass "Verblud", sampling and camping area, (H) foothills of the Susunay mountain range, the Erman's birch groove. Personalities: (C) S. Gorokhova, (F) V. Sheiko, (H) N. Kirichenko (the photographs are published with the permission of SG, VS and NK). (PK) Primorskii Krai, (SO) Sakhalinskaya Oblast (includes Sakhalin Island and Kuril Islands).

FIGURE 9 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 9. Number of gracillariid species found on various plant plants in the Russian Far East in 2010–2017. In some cases, the same gracillariid species were found on more than one plant genus (for example, Gracillaria sp. on Syringa and Fraxinus, Phyllonorycter pastorella on Salix and Populus etc.), therefore the sum of all bars is more than the total number of species identified in the study.

FIGURE 5 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 5. Female genitalia of gracillariids sampled in the Russian Far East. (A) Micrurapteryx caraganella, host plant Caragana arborescens, Skovorodino, Amurskaya Oblast, 26.VI.2016; genitalia slide [39-female]; (B) Phyllonorycter nipponicella, Quercus mongolica, PK, Gornotaezhnoe, 23.VII.2016, [8-2016-female]. Scale bars: (A) 550, (B) 100 µm.

FIGURE 6 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 6. Male genitalia of Caloptilia and Phyllonorycter spp. from the Russian Far East. (A) Caloptilia gloriosa, host plant Acer pseudosieboldianum, genitalia slide [13-male]; (B) Phyllonorycter cavella, Betula dahurica, [11-2016-male]; (C) Ph. japonica, Corylus mandshurica, [4-2016-male]; (D) Ph. jozanae, Crataegus sp. [14Pj-2016-male]; (E) Ph. issikii, T. mandshurica, [NK596-male]. Sampling locations: PK, Gornotaezhnoe, (A–D) 22–25.VII.2016, (E) 21.VIII.2015. Scale bars: (A) 350, (B–E) 200 µm.

FIGURE 14 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 14. Mines, leaf shelters and pupation sites of Phyllonorycter, Chrysaster and Cameraria spp. from the Russian Far East. (A) Phyllonorycter sp. 7, host plant Ulmus glabra; (B–C) Phyllonorycter sp. 8, Juglans mandshurica; (D–E) Chrysaster hagicola, Lespedeza bicolor; (F–G) Cameraria niphonica, Acer pseudosieboldianum and A. caudatum subsp. ukurundense, respectively; (H) Phyllocnistis sp. 1, Salix sp. Indications: (m) mine; (b) blotch part of the mine; (t) epidermal tunnel; (l) larva; (p) pupation site. Close up: (C–G) mine; (H) pupation site. Sampling locations: (A) SO, Sakhalin Isl., Yuzhno-Sakhalinsk, 11–20.VII.2017; (B–H) PK, Gornotaezhnoe, MTS and forest, 22–26.VII.2016.

FIGURE 10 in Exploring species diversity and host plant associations of leaf-mining micromoths (Lepidoptera: Gracillariidae) in the Russian Far East using DNA barcoding

FIGURE 10. Mines, leaf shelters and pupation sites of Micrurapteryx and Caloptilia spp. from the Russian Far East. (A–B) Micrurapteryx caraganella, host plant Caragana arborescens; (C) M. gradatella, Vicia sp.; (D) Caloptilia acericola, Acer pseudosieboldianum; (E) C. alni, Alnus hirsuta; (F) C. betulicola, Betula platyphylla; (G) C. heringi, A. pictum; (H) C. stigmatella, Salix sp. Indications: (m) mine; (t) epidermal tunnel; (p) pupation site; (f) folded leaf margin or leaf tip; (r1–r2) rolled leaf margin or leaf tip (the indexes 1 and 2 indicate the order of construction appearance); *mine of Diptera. Close up: (C, G, H) mine. Sampling locations: (A–C) AO, Skovorodino, 26.VI.2016; (D) PK, Gornotaezhnoe, MTS and forest, 22–27.VII.2016; (E–H) SO, Sakhalin Isl., Yuzhno-Sakhalinsk, 11–20.VII.2017.

FIGURE 7 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 7. Natural history Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) on Eugenia uniflora L. (Myrtaceae). A, host plant (mines on leaves are indicated by closed arrows); B, C, young and matures mines, respectively, on adaxial surface of leaves (larvae inside mines are pointed by open arrows); D, penultimate larval instar on dissected, mature mine, dorsal; E, last larval instar on dissected shield, dorsal; F, detached shield and corresponding orifice (indicated by closed arrow) left on leaf mine corner, where corresponding cuts were performed by penultimate instar, dorsal; G, pupa dissected from shield, near adult emergence, dorsal; H, pupal exuvium, left attached to the shield after adult emergence, lateral. Scale bars = 1 (B), 5 (C), 0.5 (D, G–H), 1 mm (E–F).

FIGURE 8 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 8. Evolutionary analysis of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) through maximum likelihood inference. The consensus tree was reconstructed based on 658 base pairs of the Cytochrome oxidase I (COI) gene. The numbers adjacent to the branches represent bootstrap support, with an asterisk denoting values below 50.The scale bar indicates the substitution rate per nucleotide site.

FIGURE 6 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 6. Scanning electron micrographs of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) pupa. A, B, head under lateral and ventral views, respectively; C, clypeal setae, ventral (detail of area indicated by square in B); D, mesothorax, dorsal; E, abdominal segment Ab, lateral; F, abdominal spiracle (detail of area indicated by square in E); G, distal portion of 5 abdomen, lateral; H, terminal hooks, lateral (detail of area indicated by square in G); I, distal portion of abdomen, ventral. Scale bars = 100 (A–B, D–E, I), 10 (C, F, H), 50 μm (G).

FIGURE 5 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 5. Scanning electron micrographs of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) last larval instar. A, B, head, under lateral and ventral views, respectively; C, antenna, ventral; D, labrum, ventral; E, spinneret, ventral (aperture indicated by closed arrow); F, maxila, ventral; G, prothoracic spiracle, lateral; H, mesothoracic calosity, ventral; I, last abdominal segments, lateral. Scale bars = 50 (A), 100 (B, I), 20 (C–D, G–H), 10 μm (E–F).

FIGURE 4 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 4. Scanning electron micrographs of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) penultimate larval instar. A–C, head, under dorsal, ventral and lateral views, respectively; D, labrum, dorsal; E, spinneret (indicated by closed arrow), ventral; F, antenna, ventral; G, prothoracic spiracle, lateral; H, abdominal segment Ab3, lateral; I, last abdominal segments, lateral. Scale bars = 100 (A–C, H–I), 20 (D), 10 μm (E–G).

FIGURE 2 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 2. Morphology of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) reproductive structures under light microscopy. A, male genitalia, ventral view; B, wings, dorsal; C, transtilla, ventral; D, phallus, lateral; E, left valva, ventral (closed arrow indicates pectinifer); F, female abdomen with retracted ovipositor (closed arrow points to spermatheca), lateral; G, distal portion of female abdomen with protracted ovipositor (indicated by open arrow), lateral. Scale bars = 50 (A, D), 100 (E), 500 (B), 30 (C), 200 μm (F–G).

FIGURE 1 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 1. Adults of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae). A, male (holotype), pinned-dried, dorsal view; B, corresponding forewing posterior margin in detail; C, D, female on Eugenia uniflora L. leaf, under lateral and dorsal views, respectively. Scale bars = 1 (A, C, D), 0.5 mm (B).

FIGURE 3 in Heliozela pitangavora Moreira & Fochezato sp. nov. (Lepidoptera: Heliozelidae): a leaf miner micromoth associated with Eugenia uniflora L. (Myrtaceae) in southern Brazil

FIGURE 3. Immature stages of Heliozela pitangavora sp. nov. (Lepidoptera: Heliozelidae) under light microscopy. A, penultimate larval instar (dorsal and ventral, respectively); B, last larval instar (dorsal and ventral); C, pupa (dorsal, ventral, and lateral). Scale bars = 500 μm.