The family Psychidae is consisting of 241 genera and 1,350 species (Sobczyk, 2011; van Nieukerken et al., 2011). Among then, the subfamily Oiketicinae was established by Herrich-Schäffer in 1855 based on the type genus, OiketicusGuilding, 1827. The Oiketicinae species have a relatively large-sized, consisting more than 388 described species under 4 tribe worldwide (Sobczyk, 2011).
The genus Nipponopsyche belongs to Oiketicinae, based on the type species Nipponopsyche fuscescensYazaki, 1926. In the world, the genus Nipponopsyche has been known only one species, which was distributed in Japan (Sauter and Hättenschwiler, 1991; Sobczyk, 2011;Saigusa and Sugimoto, 2013). Yazaki (1926) diagnosed the genus Nipponopsyche with the combination of the following adult characters: antennae bipectinate and scape very large, forewing very broad with nine veins arising from discal cell. As other species of Psychidae, larvae of the genus tend to build their cases putting together feed-grasses. The genus Nipponopsyche mainly live in low-elevation grassland, wasteland or wetland in their larval stage. However, some habitat has been reduced by the use of power brush cutter recently (Saigusa and Sugimoto, 2013).
In the present study, N. fuscescens is newly recognized from South Korea for the frist time. Also, all the available information is presented, including the collection locations, microhabitats, and illustrations of larva, pupa, adults and its genitalia with SEM. DNA barcode is also provided for precise identification of the species.
Materials and Methods
The materials examined in this study are based on the Entomological Collection of the Korea National Arboretum, Pocheon, Korea (KNAE). Specimens were dissected and examined after mounting on slide glass; male genitalia and wing scales in 80% glycerol solution and wing venation on dried condition. Photographs of adults and genitalia were taken using a DFC 495 digital camera attached to Leica M205A stereomicroscope (Leica, Wetzlar, Germany). Also, drawing of the wing venation was taken using a Adobe Illustrator CS5 and Adobe Photoshop CS6 (Adobe systems, San Jose, USA). To examine some morphological character of larvae, scanning electron microscopy (SEM) was used. For the dried specimen, the larvae dehydrated on 50-99% ETOH in order. And then, critical poing dryer (EMITECH K850, Ashford, Kent, UK) was used for complete dehydration. Each specimens were gold coated using Ion Sputter E-1010 (HITACHI, Tokyo, Japan) and examined with S-3400N (HITACHI, Tokyo, Japan) electron scanning microscope.
Terminology and morphological characters follows Kristensen (2003), Sugimoto and Saigusa (2001), Kozhanchikov (1956).
Genomic DNA was extracted from the legs of dried specimen for males and thorax parts of immersion specimens for females and larva, preserved in 100% alcohol using a Genomic Cell/Tissue Spin Mini Kit (Qiagen, Inc, Hilden, Germany), according to the manufacturer’s protocol. A total of three specimens were sequenced for 658 bp fragment of the mitochondrial cytochrome c oxidase I (COI) gene, and the DNA barcode was amplified using the primer LepF1, LepR1 (Hebert et al., 2004). PCR conditions for amplification followed the manufacturer’s protocol (Platinum Taq, Invitrogen, Carlsbad City, CA, USA). The amplicons were purified using the QIAquick® PCR purification kit (QIAGEN, Inc, Hilden, Germany) and directly sequenced at Macrogen (Seoul, Korea). Contigs were assembled using CodonCode aligner version 2.0.6 (CodonCode Co., Centerville City, MA, USA). Successful sequences were uploaded to BOLD systems (BOLD BIN no. BOLD:ADJ9301).
Taxonomic accounts
Genus NipponopsycheYazaki, 1926 잔디주머니나방속(신칭) NipponopsycheYazaki, 1926: 173, 174.
Type species:Nipponopsyche fuscescensYazaki, 1926: 174 by monotypy.
Nipponopsyche fuscescensYazaki, 1926 잔디주머니나방(신칭) Nipponopsyche fuscescensYazaki, 1926: 174. Type locality: Japan.
Adult. Male (Fig. 1). Wingspan 17.3-21.2 mm. Coloration and vestiture: sclerites on head and thorax dark-brown. Head (Fig. 1D and E) clothed with slightly long light brown hairs dorsally and anteriorly. Compound eyes blackish. Thoracic notum densely clothed with long brown hairs downwardly. Forewings: ground color light brown; generally covered brown scales. Hindwing covered with brown scales, basal part slightly short brown hairs. Legs (Fig. 1F) red-brown on coxae, femora and tibiae; coxae and femora covered with brown long hairs; tarsi covered with short light brown hairs; claws reddish brown. Structure: head including eyes nearly 2/5 as wide as thorax in dorsal aspect; mouthparts mostly reduced. Ocelli absent. Antenna (Fig. 1C): basal flagellomeres 29 segmented, pectinate, slightly longer than 1/2 forewing, length of antenna nearly 3.4-3.7 mm, flagellomeres covered with dark-brown scales. Forewing (Fig. 1B) short and wide, nearly triangular, costa straightly, then gently arched beyond 4/5, apex very obtuse, posterior margin straight downwardly. Scales (Fig. 1G) very slender, truncate apical margin usually produced into 1-3 weak rounded laciniation. 9 veins arising from discoidal cell; discoidal cell longer than 0.67 times as long as wing, distal corner of anterior part rectangular; Sc reaching to 3/5 costa; R1 and R2 originated at anterior part of discoidal cell; R3 and R4 stalked at apical corner of anterior part of the discoidal cell; R3 reaching to the near apex; R4 arising to 2/5 R3; M1 absent; M2 and M3 originated at apical corner of posterior part of discoidal cell; CuA1 and CuA2 close to distal corner of posterior part of discoidal cell; CuP weak; A1+A2 straight at hind margin. Hindwing (Fig. 2) fan shape. Scales (Fig. 1H) slender with arrowhead shape. Costa slowly curved, discoidal cell 3/5 length of hindwing with asymmetrical, gradually dilating apically; posterior part of discoidal cell longer than anterior part; Sc+R1 reaching to 4/5 of costal margin; Rs ending at apex; M1 absent; M2 and M3 originated at distal corner of posterior part of discoidal cell and parallel; CuA1 arising to 1/3 M3; CuP straight to termen.
Male genitalia (Fig. 3). Genitalia coverd with long 8th abdominal segment. In lateral aspect, dorsum nearly curved; saccus very long; ampulla club shape with setae sparsely; harpe hooked apically. In dorso-ventral aspect, uncus very weak concave, tegumen broad, valva short and apex rounded, vinculum wide and rapidly arched to saccus, saccus slender and elongated downwardly, phallus shorter than genitalia.
Female (Fig. 2). Length 12.5-13.9 mm, vermiform, cylindrical, prothorax, mesothorax and metathorax thin, almost same thickness from the 3th to 6th abdominal segment, 8+10th abdominal segments with short papillae anales. Coloration: head brown with matt vertex. Meso and metanotum brown. Membranous areas of abdomen yellowish. Structure: head (Fig. 2D and E) reduced in size, about 0.05 times as long as body length; mouth part considerably degenerated, antennae very short, eye-spots blackish. Thorax very small and 0.07 times as long as body length, dorsal margin slightly curved and covered with few scales, prothorax narrower than meso and metathorax. Legs (Fig. 2G) almost degenerated with very short. Abdomen well developed and 0.82 times as long as body length, first abdominal segment weak sclerotized, second to 6th abdominal segments almost membranous, 7th abdominal segment approximately 1.2 times as long as the 6th segment, 7th segment (corethrogyne or anal hair tuft) covered with few hairs, eighth abdominal segment very short, 9th and 10th abdominal segments present papillae anales (ovipositor), papillae anales very short.
Larva (Fig. 4). Length 14-16.3 mm. Coloration and vestiture: head present thick black lines, epicranial suture black, frons pale yellow, clypeus and labrum dark brown. Dorsal aspect of thorax pale yellow with thick black lines. Abdomen brown, black lines present in dorsal aspect; membranous parts and sclerites pale yellow. Thoracic legs pale yellow with black some line. Abdominal and anal proelg pale yellow. Structure: head slightly large; epicranial notch concave, epicranial suture slightly thick, adfrontal area very thick and slightly curved downwardly; antenna short; frons triangular, C1 setae coiled; clypeus protuberant weakly; labrum slightly narrow and concave at ventral aspect; mandible broad. Thoracic legs slightly thick with many setae, pretarsus hooked; prothoracic shield well sclerotized. Abdominal prolegs short. Anal shield weak sclerotized, anal proleg very short, abdominal proleg crochets U-shaped with short setae; spiracles oval shaped.
Pupae (Fig. 5). Male (Fig. 5A-C): length 10.7-11.5 mm. reddish brown on head, thorax and abdomen, brown on legs. Body cylindrical, present setae sparsely. Swollen at vertex; pronotum slightly arched; maxillae considerably thick; maxillary palps triangular shape; antennae quite wide with transverse line regularly; I and II thoracic legs slightly slender and straight downwardly. Forewing moderately wide, comprising 2/5 of pupa; hindwing narrowed to posterior margin of abdomen. posterior margin of mesonotum wide and U-shape; metanotum narrow and slighlty convex. second to 6th abdominal segments almost same thickness, 7th to 9th narrowed thickness; 9th abdominal segment slightly rounded. Anal hooked.
Female (Fig. 5D-F). Length 14-15.4 mm. Body reddish brown, covered with some scales, nearly same width and thickness as 4th abdominal segment, evenly narrowed to tip. Head very small, considerably degenerated. Thorax narrowly simple form, three thoracic segments of nearly the same length with less than 1/10 of pupa. Abdominal segments slightly tapered in basally. Anal slightly blunt.
Larval case (Fig. 7). In the observation, this Larvae build their cases by putting together lawn grasses. Also, they are live around the grave.
Material examined. 4♂, 33♀ (emerged individuals), 4 larvae, Chuja Is., Jeju Is., lat.33.96399722, long.126.2879, 30.v.2018 (S.J. Roh, D.S. Kim, B.S. Park and S.B. Choi), genitalia mounted on 80% glycerol solution, gen. no. KNAESJ00044, scales of wing mounted on 80% glycerol solution, sclaes of wing no. KNAESSJ10, wing venation no. KNAEVS18-coll. KNAE.
Distribution. Korea (new record), Japan.
DNA barcode. DNA barcode sequence were generated (BOLD Process ID. NFNK001-18, NFNK002-18, NFNK003-18). Multiple alignments using the BLAST tool in the BOLD systems database showed the following species as nearest neighbor: Nipponopsyche fuscescens (locality of reference data from Japan) 99.85%.
Remarks. Parthenogenesis is known at least four subfamilies and five genera in the Psychidae (Sobczyk 2011;Elzinga et al., 2013). In this study, we found that N. fuscescens occurrs parthenogenetic form (Fig. 6). This Larvae build their cases by putting together lawn grasses. Also, they are live around the grave (Fig. 7). Adults emerge from late June to late July in breeding condition. Out of 37 larvae, 33 females and 4 males were emerged in the present study.