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Research Article
Discovery of a new species of Douglasiidae Heinemann & Wocke, 1876 (Lepidoptera) in Japan
expand article infoSadahisa Yagi‡§
‡ Insect DX Laboratory, Faculty of Agriculture, Kyushu University, Fukuoka, Japan
§ Insect Science and Creative Entomology Center, Kyushu University, Fukuoka, Japan
Open Access

Abstract.

Tinagma nigripunctum sp. nov. is described from the Okinawajima Island, Japan. This is the first record of Douglasiidae in Japan and the Oriental Region.

Introduction

The family Douglasiidae Heinemann & Wocke, 1876 is a small family of Lepidoptera, the majority of which are distributed in the Holarctic region, particularly in Europe (Gaedike 1974; Komai et al. 2011). Two extant genera (Tinagma Zeller, 1839 and Klimeschia Amsel, 1938) and 39 extant species are recognised globally (Common 1990; Gaedike 1974, 1987, 1990, 1991, 2010, 2018, 2021; Gaedike and Falck 2019; Gaedike and Šumpich 2017; Harrison 2005; Nel and Varenne 2019), and Protonyctia originalis Meyrick, 1932 is also sometimes included in this family (Edwards and Kallies 2010; Heppner 2012). One fossil genus with one species has been described from Myanmar amber (Poinar Jr 2017). This family was previously recognised as a member of Gracillarioidea or Yponomeutoidea (Common 1970; Davis and Robinson 1998; Komai et al. 2011). However, it is currently treated as within a separate monobasic superfamily of Apoditrysia (Mitter et al. 2017).

The immature stages of this family are poorly understood. However, species of Klimeschia have been reported as leaf miners and Tinagma species as flower petiole miners and stem borers in Boraginaceae, Rosaceae, and Lamiaceae (Heppner 1987; Davis and Robinson 1998).

Although several species of this family have been recorded in the Russian Far East, no members of Douglasiidae had been discovered so far in Japan. During a field survey on Okinawajima Island, at the northernmost tip of the Oriental Region, an unknown species was collected. After careful examination, this species was determined to be an undescribed species of Tinagma, Douglasiidae. This study describes this new species, and infers its relationships, based on morphological and molecular analyses.

Materials and Methods

Materials and observations

All the specimens were collected using a light trap and deposited at the Entomological Laboratory of Kyushu University (ELKU). The morphological characteristics were observed using a Leica S9D stereomicroscope. The genitalia were dissected, stained with chlorazol black E, and mounted in Euparal. The morphological terminology follows that of Harrison (2005).

DNA analysis

Two type specimens were used for DNA analysis. DNA was extracted from the abdomen. For DNA analysis methods, see Yagi et al. (2024). The fragment of the partial mitochondrial cytochrome c oxidase subunit I (COI-5P) gene was amplified using the primer sets LCO1490/HCO2198 (Folmer et al. 1994) and the KOD One® PCR Master Mix -Blue- (TOYOBO CO., LTD., Osaka, Japan), following the manufacturer’s protocol, except that the annealing temperature was 45 °C.

The DNA barcode sequences were deposited in BOLDSYSTEMS (Ratnasingham and Hebert 2007) in the dataset DS-TINAJPN (https://doi.org/10.5883/DS-TINAJPN). The sequences were also deposited in GenBank under the accession numbers: PX924406 and PX924407. Sequences for all Tinagma species were downloaded from BOLDSYSTEMS (Table 1). Intraspecific uncorrected pairwise distances (p-distances) were calculated using MEGA 7.0.26 (Kumar et al. 2016). For visualisation, a neighbour-joining tree was constructed with the uncorrected pairwise distances method using MEGA 7.0.26 (Kumar et al. 2016), with bootstrap analysis performed with 1000 repetitions. The treefile was visualised using Figtree v1.4.4 (Rambaut 2018).

Table 1.

Sample information of Tinagma used for DNA analysis.

Species sample ID Process ID location BIN ID GenBank accession number References
Tinagma nigripunctum sp. nov., holotype SaY1342 DOUJP001-25 Japan, Okinawa Pref. BOLD:AGT4721 PX924407 This study
Tinagma nigripunctum sp. nov., paratype SaY1343 DOUJP002-25 Japan, Okinawa Pref. BOLD:AGT4721 PX924406 This study
Tinagma anchusella TLMF Lep 06609 PHLAH109-12 Austria, Lower Austria BOLD:ABW5879 – Mutanen et al. (2016)
Tinagma balteolella INV00587 IBILP1813-20 Portugal, Braganca BOLD:AAZ0894 OQ562379 Ferreira et al. (2024)
Tinagma californicum Gaedike, 1990 CNCLEP00020403 LNEL083-06 United States, Washington BOLD:AAK1547 – –
Tinagma dryadis Staudinger, 1872 TLMF Lep 02763 PHLAC728-10 Switzerland, Graubünden BOLD:AAB3291 JF860280 Mutanen et al. (2016)
Tinagma giganteum Braun, 1921 CNCLEP00026491 MNAA337-07 Canada, Yukon Territory BOLD:AAG2052 – –
Tinagma jalavai Gaedike, 2018 CCDB-23847 F11 MICRR261-18 Russia, Altai Republic BOLD:ACF9705 – –
Tinagma leucanthes Meyrick, 1897 11ANIC-15698 Australia, Tasmania BOLD:AAY8145 KF389562 Hebert et al. (2013)
Tinagma mongolicum Gaedike, 1991 KLM Lep 06384 LEALT779-16 Russia, Altai Republic BOLD:ADE9335 – –
Tinagma obscurofasciella (Chambers, 1881) CNCLEP00146032 MNAM1136-16 Canada, Quebec BOLD:ACF8424 – –
Tinagma ocnerostomella (Stainton, 1850) MM00483 LEFIB122-10 Finland, Finland Proper BOLD:AAI0333 HM871028 Mutanen et al. (2016)
Tinagma perdicella Zeller, 1839 MM14142 LEFIG233-10 Finland, Ostrobottnia BOLD:AAI0292 HM875912 Mutanen et al. (2016)
Tinagma pulverilinea Braun, 1921 jflandry1622 MECB563-05 Canada, Alberta BOLD:AAI0294 – –
Tinagma signatum Gaedike, 1991 TLMF Lep 10252 LEATB075-13 Austria, Tyrol BOLD:ACE2011 – Huemer and Hebert (2016)
Klimeschia transversella (Zeller, 1839) MM08531 LEFIE216-10 Finland, Regio aboensis BOLD:AAG2055 HM873953 Mutanen et al. (2016)

Taxonomy

Tinagma nigripunctum sp. nov.

Figs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17

Japanese name.

kurohoshi-hime-moguriga.

Figures 1–4. 

Adult habitus of Tinagma nigripunctum sp. nov. 1, 3. Male, holotype; 2, 4. Female, paratype; 1, 2. Dorsal view; 3. Diagonally forward right view; 4. Lateral view.

Figures 5, 6. 

Male abdominal segments of holotype Tinagma nigripunctum sp. nov. 5. First and second segment; 6. Eighth segment.

Figures 7–10. 

Male genitalia of holotype Tinagma nigripunctum sp. nov. 7. Complete genitalia except phallus; 8. Phallus; 9. Tegumen; 10. Vinculum.

Figures 11, 12. 

Male terminal segments of holotype Tinagma nigripunctum sp. nov. 11. Lateral view; 12. Ventral view.

Figures 13–16. 

Female genitalia of paratype Tinagma nigripunctum sp. nov. 13. Complete genitalia. 14. Ostium bursae. 15. Opening of ductus seminalis. 16. Signum.

Figure 17. 

Neighbor Joining tree of the COI barcode region from the Tinagma species based on uncorrected pairwise distances. Numbers near the nodes indicate the bootstrapping supports.

Type material.

Holotype. male. Japan: Okinawa Pref. Itokazu-joato, Itokazu, Nanjo, 24.III.2021 LT, S. Yagi, J. Oku, HK. Kim, K. Goto, M. Kimura leg., genitalia slide no. SY1638, Sample ID: SaY1342.

Paratype. 1 female, same data as holotype, genitalia slide no. SY1639, Sample ID: SaY1343.

Diagnosis.

Most Tinagma species are difficult to distinguish based on external morphology, as their forewings are greyish brown with white to grey markings. However, this species can be distinguished from others by a combination of characters, particularly an indistinct pale greyish-brown fascia, a fuscous spot at the apex, and small body size. In the male genitalia, this species is similar to T. balteolella (Fischer von Röslerstamm, 1840), T. klimeschi Gaedike, 1987, and T. anchusella (Benander, 1936) in sharing a costal process and an articulated process in the valva. However, it can be distinguished by the acute tip of the valva and the uninterrupted pecten on the outer margin of the valva (roundish tip and interrupted pecten in T. balteolella and T. klimeschi, and roundish tip without pecten in T. anchusella). In the female genitalia, this species is similar to T. balteolella and T. klimeschi in having an elongated sterigma with a U-shaped emargination on the ventral opening. However, it can be distinguished by the straight and uniform width of the sterigma (bulging in T. balteolella and T. klimeschi).

Description.

Male (Figs 1, 2, 3, 4, 5, 6). Wingspan: 5.5 mm in holotype. Forewing length: 2.5 mm in holotype. Head: smooth, greyish brown. Frons greyish cream. Ocellus well developed. Antenna greyish brown, filiform but slightly thickened, 0.65× length of forewing. Proboscis long, without scales. Maxillary palpus reduced. Labial palpus pale brown, short, straight, apically dark greyish-brown. Thorax and tegula: dark greyish brown. Forewing upper side greyish brown with faint pale greyish brown fascia in the middle, anteriorly bordered by dark brown band; apex with fuscous spot; fringe pale brown; underside greyish brown, costa basal half dark brown. Hindwing dull brown; fringe pale brown. Frenulum a single stout seta. Foreleg epiphysis present. Legs greyish brown; each tarsomere dark greyish brown, ringed apically. Abdomen: greyish beige dorsally, beige ventrally. S2 with slender apodeme, curved inwards; venula present (Fig. 5). S8 bar-shaped, strongly sclerotised, laterally with a pair of slender rods projected inward. T8 weakly sclerotised, trapezoid, posteriorly slightly concave (Fig. 6).

Female. Wingspan: 5.9 mm in paratype. Forewing length: 2.8 mm in paratype. Similar to male but antenna filiform, 0.5× length of the forewing. Frenulum comprising 2 setae.

Male genitalia (Figs 7, 8, 9, 10, 11, 12). Uncus and gnathos absent. Tegumen slightly sclerotised, triangular, ventrally with microtrichia and subapically with two pairs of long setae (Fig. 9). Valvae symmetrical; trapezoid valva with short, obliquely upward costal projection; apex of valva with an acuminate articulated process 5× as long as basal width, medially folded over valval surface and anteriorly projecting to 0.7× length of valva; ventral margin nearly straight with setose fold; outer margin concave with uninterrupted pecten comprising ca. 18 thickened setae; dorsal margin smooth. Transtilla slender, tapered toward midline. Vinculum a strongly sclerotised band with triangularly projecting lateral lobe; saccus moderately projecting; median membranous area of vinculum with minute microtrichia (Fig. 10). Anellus a sclerotised ring, subapically angular, apically rounded and projecting. Juxta basally flat with broadly sclerotised plate. Phallus long, slender, gently tapered toward tip, basally curved upward; coecum penis small; cornuti absent.

Female genitalia (Figs 13–16). Papillae anales moderately long. Apophyses posteriores slender, 1.5× length of apophyses anteriores. Apophyses anteriores connected to a setose triangular lateral sclerite posteriorly. S9 triangular, with minute spines. Posterior margin of S8 serrated and almost straight. Ostium bursae arising from the membrane between eighth and ninth sterna, strongly emarginate. Sterigma strongly sclerotised, elongated ring, uniform in width, ventral opening U-shaped and serrated. Ductus bursae slender; slightly constricted posteriorly; posterior 3/5 weakly sclerotised, anterior 2/5 membranous with minute pectinations; anterior border of sclerotised region with six small sclerotised spinules. Ductus seminalis arising at posterior 3/5 of ductus bursae near spinules. Corpus bursae oval, with signum; signum consisting of numerous strong sclerotised slender rods of various sizes, arranged in a radial pattern; minute pectinations around signum.

DNA data

(Fig. 17, Table 2). The COI barcode sequences (658 bp) of one male and one female Tinagma nigripunctum sp. nov. 100% matched. The DNA barcode of Tinagma nigripunctum sp. nov. (BOLD:AGT4721) is closest to that of T. balteolella (BOLD:AAZ0894), with a pairwise distance (p-distance) of 9.7%. The NJ tree also showed similarity to T. anchusella (BOLD:ABW5879), with a pairwise distance of 10.6%.

Table 2.

The uncorrected pairwise distances of Tinagma species based on COI barcode region.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
1 SaY1342_Tinagma_nigripunctum_holotype_PX924407
2 SaY1343_Tinagma_nigripunctum_paratype_PX924406 0.000
3 IBILP1813-20_Tinagma_balteolella_OQ562379 0.097 0.097
4 LNEL083-06_Tinagma_californicum 0.102 0.102 0.097
5 PHLAH109-12_Tinagma_anchusella 0.106 0.106 0.087 0.113
6 LEFIB122-10_Tinagma_ocnerostomella_HM871028 0.111 0.111 0.106 0.097 0.099
7 MICRR261-18_Tinagma_jalavai 0.112 0.112 0.091 0.093 0.108 0.097
8 LEFIG233-10_Tinagma_perdicella_HM875912 0.116 0.116 0.094 0.102 0.111 0.106 0.059
9 MNAA337-07_Tinagma_giganteum 0.116 0.116 0.097 0.085 0.110 0.094 0.019 0.063
10 PHLAC728-10_Tinagma_dryadis_JF860280 0.117 0.117 0.107 0.098 0.107 0.096 0.055 0.043 0.057
11 MNAM1136-16_Tinagma_obscurofasciella 0.119 0.119 0.090 0.096 0.108 0.100 0.024 0.059 0.027 0.047
12 MECB563-05_Tinagma_pulverilinea 0.123 0.123 0.106 0.104 0.109 0.112 0.055 0.067 0.052 0.062 0.053
13 LEALT779-16_Tinagma_mongolicum 0.128 0.128 0.108 0.110 0.116 0.119 0.058 0.061 0.052 0.064 0.050 0.056
14 LEATB075-13_Tinagma_signatum 0.129 0.129 0.114 0.107 0.125 0.109 0.068 0.067 0.071 0.053 0.067 0.070 0.082
15 ANICX1698-11_Tinagma_leucanthes_KF389562 0.157 0.157 0.169 0.155 0.161 0.134 0.164 0.175 0.164 0.163 0.155 0.158 0.164 0.179
16 LEFIE216-10_Klimeschia_transversella_HM873953 0.116 0.116 0.119 0.129 0.122 0.117 0.111 0.108 0.113 0.110 0.111 0.122 0.108 0.120 0.161

Biology.

Unknown (see the discussion below for the suspected host plant family). Adults were collected at the forest edge in late March (Fig. 18).

Figure 18. 

Type locality of Tinagma nigripunctum sp. nov. Forest edge, Okinawajima Island, Japan.

Etymology.

The name of the new species, an adjective, is derived from the Latin nigrum, “black” and punctum, “spot”, referring to the black dot at the apex of the forewing.

Remarks.

This species is one of the smallest species of Douglasiidae, along with Tinagma minutissima (Staudinger, 1880) (wingspan 4.5–6.5 mm; Budashkin 2003).

Discussion

Based on the genetic distances, this new species shows greater genetic divergence from the other Holarctic species included in this study (9.7–15.7%) than the genetic divergence observed among those species (2.7–12.5%, Table 2). This pattern may indicate a long period of isolation and is consistent with a geographically distant distribution relative to the other Holarctic taxa.

Based on the preliminary phylogenetic tree and genetic distances, this species is similar to T. balteolella and T. anchusella although the bootstrap values are low. This similarity is also supported by the genital morphology, with the shared presence of a single costal process on each valva. The host plant of the new species remains unknown; however, both T. balteolella and T. anchusella are known to use herbaceous Boraginaceae (Gaedike 1974). Therefore, it is possible that this new species also feeds on Boraginaceae. In April 2025, the herbaceous Boraginaceae were surveyed at the type locality but no candidate host plants were discovered.

This new species is the first record of an extant species of this family from the Oriental Region. A fossil species has been recorded from amber in Myanmar (Poinar Jr 2017). However, this fossil species does not appear to belong to Douglasiidae because it has several characteristics not observed in Douglasiidae, including the development of five palpomeres on the maxillary palpus, slender hair-like scales on the vertex and bristle-like hairs on the underside of the labial palpus. These characteristics and wing shapes are often seen in Tineoidea, for example, Tineidae and Meessiidae (Davis and Robinson 1998; Regier et al. 2015), but the type specimen was not checked, and further determination of the family placement is still pending.

It is difficult to speculate on the origin of this species, but the reason why this species has not been found until now is thought to be that it was overlooked because of its small size rather than because it was recently introduced. Based on the occurrence data of this genus from GBIF (http://id.luomus.fi/F.462239), a single female specimen collected in Thailand was identified as a species of Tinagma by Dr Gaedike, an authority on the taxonomy of this family. This record and the new species described here suggest that species of this family also inhabit other localities in the Oriental Region. Similar underestimation of microlepidopteran species diversity is documented in many groups, such as Nepticulidae and Lecithoceridae, because the study of micromoths in the Oriental Region remains in its infancy despite recent progress (e.g., van Nieukerken et al. 2016; Park 2022). Further discoveries are expected through future field surveys (Fig. 18).

Acknowledgements

I thank Mr Masaaki Kimura (Okinawa Prefecture), Dr Johei Oku (Oita Prefecture), Dr Hyeong Kyu Kim (Akita Prefectural University), and Mr Kiyoshiro Goto (Kumamoto Prefecture) for their support during the field survey on Okinawajima Island in 2021. I also thank Mr Jinhyeong Park (Kyushu University) for his advice on the morphology of Tineoidea. I sincerely thank Dr Sergey Sinev and Dr Vladimir Lukhtanov (Zoological Institute of Russian Academy of Sciences) for permission to use DNA barcode data deposited in BOLD Systems. I am grateful to Dr Toshiya Hirowatari for his valuable comments regarding this manuscript. Finally, I gratefully acknowledge the support of the SEL for covering the costs of processing the article (APCs).

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