A new species of Pima Hulst, 1888 from China (Lepidoptera, Pyralidae, Phycitinae), with a key to Holarctic species

Abstract Pima tristriatasp. nov. is described as new to science based on specimens collected from the Ningxia Hui Autonomous Region, China, and P. boisduvaliella (Guenée, 1845) is also treated here for comparison. DNA barcodes of the two species are provided, together with a neighbor-joining tree for species delimitation. A key to the Holarctic species and a distribution map of the Chinese species are presented.


Introduction
The genus Pima was established by Hulst (1888) with Pima fosterella Hulst as the type species. Ragonot (1889Ragonot ( , 1893 referred P. fosterella and the other congeneric American species to Epischnia Hübner, 1825. Heinrich (1956) revised the genus Pima from America, pointing out that Epischnia as defined by Ragonot was a composite of several disparate species and that none of them agreed with the type species of Epischnia, and transferred eight species to Pima. Neunzig (2003) treated nine species of Pima from North America, described one new species and proposed two synonyms. Leraut (2014) treated five species from Europe, including one new species and two new combinations. Tsvetkov (2016) described P. transfusor Tsvetkov from the South Urals. Moreno and Gastón (2017) transferred four species to Pima. Falck et al. (2019) described P. tricolorella Falck, Karsholt & Slamka from the Canary Islands of Spain. Slamka (2019) reviewed the genus in Europe, synonymized Palloria Amsel with Pima and Pima leucomixtella (Ragonot) with Pima christophori (Ragonot), transferred Epiepischnia keredjella Amsel and Epischnia trifidella Zerny to Pima, and described three new species. Twenty-four species have hitherto been assigned to Pima worldwide, mainly distributed in North America and Europe.
Two species, Pima boisduvaliella (Guenée) and P. trifidella (Zerny) were reported from China before this study. We herein describe one new species, Pima tristriata sp. nov., provide DNA barcodes of the new species and P. boisduvaliella (Guenée), and a neighbor-joining tree covering seven species for species delimitation. A key to the known Holarctic species of the genus Pima is also provided.

Materials and methods
The examined specimens in this study were collected by light traps in the Ningxia Hui Autonomous Region, China. Morphological terminology follows Heinrich (1956). Genitalia and wings were dissected and mounted according to the methods introduced by Li (2002). Illustrations were prepared using a Leica DM750 microscope, and refined in Photoshop ® CS4 software. Photographs of adults were taken with a Leica M205A stereo microscope. The cartographic illustration was made using DIVA-GIS 7.5 (Hijmans et al. 2005). All specimens examined, including the holotype of the new species, are deposited in the Insect Collection of Nankai University, Tianjin, China (NKU).
DNA was extracted from dry adult specimens using Qiagen ® DNeasy Blood & Tissue Kit, with the genitalia mounted on slides as vouchers. Samples were amplified using the primers LCO1490 and HCO2198 (Folmer et al. 1994) in 25 μl reaction volume: 0.75 μl of each primer (10 mM; Sangon Biotech), 2 μl DNA template, 12.5 μl mixture (KOD One PCR Master Mix; TOYOBO), and 9 μl ddH 2 O. PCR reaction conditions used were as follows: 35 cycles of 98 °C for 10 s, 55 °C for 5 s, 68 °C for 1 s; then a 4 °C hold. A weak electrophoretic band of the new species was obtained, and the PCR product was recovered (SanPrep Column DNA Gel Extraction Kit; Sangon Biotech) and cloned (Hieff CloneTM Zero TOPO-TA Cloning Kit; Sangon Biotech). Positive plasmids were sent to Sangon Biotech (Shanghai, China) for sequencing.
Genetic distance estimation and neighbor-joining analysis were conducted in MEGA X using the Kimura 2-Parameter model. Thirty-eight sequences were used in the analyses: one new sequence from a paratype of P. tristriata sp. nov. (GenBank accession number MT749678) and three new ones from Chinese specimens of P. boisduvaliella (GenBank accession numbers MT734539, MT734540, MT734541), the others from BOLD (Ratnasingham and Hebert 2007). The extreme values of the interspecific and intraspecific distances were presented in Table 2, and the NJ tree was shown in Fig. 8 Amsel, 1961 Diagnostic characters. Pima is characterized by the male basal few flagellomeres shallowly incurved and containing a row of minute, tooth-like spines (Figs 1a,2a), third segment of the labial palpus projected forward (Figs 1b,2b); the forewing usually having a white subcostal streak (absent in P. keredjella, P. milka, P. parkerella, P. pempeliella, P. transfusor and P. tristriata sp. nov.), with 11 veins (Figs 1c, 2c), R 2 approximate to the stalk of R 3+4 + R 5 , R 3+4 stalked with R 5 of less than half their lengths, M 2 , M 3 and CuA 1 free; the hindwing with 10 veins (Figs 1c, 2c), Rs and M 1 shortly stalked, M 2 and M 3 stalked for over half their length, CuA 1 and M 2 + M 3 shortly stalked; apical process of gnathos short and stout, transtilla absent, the broad costa of the narrowed valva with a blunt, slightly forked apex (more pointed and not forked in P. christophori, P. leucoloma, P. pempeliella, and P. trifidella), the uncus with a broad base and a short pair of lateral lobes, the aedeagus with two stout cornuti (one cornutus in P. trifidella) in male genitalia (Figs 3,4); the ductus bursae ribbon-like, the stout corpus bursae scobinategranulate and usually with sclerotized patches or folds in female genitalia (Figs 5,6).
Pima resembles Epischnia Hübner, but they can be separated by the following characters: in Pima, the male flagellum with a row of tooth-like spines near the base, the labial palpus with terminal two segments approximately of equal length; male genitalia with a broad, apically slightly forked costa, and two stout cornuti in the aedeagus; female genitalia with a strongly sclerotized, funnel-shaped antrum, the corpus bursae scobinate-granulate throughout and with sclerotized patches or folds. Whereas, in Epischnia, the male flagellum lacks a tooth-like spine, the third of the labial palpus is less than half the length of the second; the costa is weak and not forked at the apex, and the aedeagus has a bunch of spinules in the male genitalia; the antrum is weak or represented by a band-shaped plate, the corpus bursae is smooth on the inner surface except for one big sclerotized plate or a line of small thorns and one bunch of spinules in the female genitalia.  (Roesler 1990: fig. 9); apophyses posteriores slightly shorter than anteriores (Roesler 1990: fig. 10 (Heinrich 1956: fig. 306); corpus bursae more rounded, with a projecting shield at junction with ductus bursae (Heinrich 1956: fig. 782)  cell respectively, whereas, most of the other congeners have a white subcostal streak. It is superficially similar to P. parkerella (Schaus), but with differences in genitalia: juxta with globular lateral lobes, costa projected beyond apex of valva, and corpus bursae heart-shaped in the new species; juxta with short finger-like lateral lobes, costa terminated at end of valva, and corpus bursae rounded in P. parkerella. It resembles Pima boisduvaliella (Guenée) in genitalia except for some slight differences: lateral lobes the juxta is globular, the vinculum is ca 2× length of its greatest width, the aedeagus is approximately equal to valva in length in the male genitalia, and the corpus bursae has an irregular sclerotized plate in the female genitalia. In P. boisduvaliella, lateral lobes the juxta is slender, finger-like, the vinculum is ca 1.5× length of its greatest width, and the aedeagus is 1.2× length of valva in the male genitalia; the corpus bursae has a couple of tortuous, sclerotized plates in the female genitalia.
Description. Adult (Fig. 1). Wingspan 25.5-31.0 mm. Head (Fig. 1a, b) grayish white. Antenna grayish white, scape ca 1.5× as long as wide, flagellum of male with short cilia, of female pubescent. Labial palpus of male grayish white mixed with a few brown scales, of female brown mixed with a few grayish white scales; first and second segments obliquely upturned, third second projected forward; third segment as long as second, twice as long as first. Maxillary palpus minute, grayish brown, in form of an aigrette. Patagium, tegula and thorax pale yellow, mottled a few brown scales. Forewing yellow, costa dorsum and lower margin of cell overlaid with a longitudinal grayish black streak respectively, more or less peppering of whitish scales; some scattered black dotting along veins and termen; antemedial line white, arched, white, from costal 1/5 slightly oblique to dorsum 1/4, inner bordering ashy black on lower half, outer edging of grayish brown; postmedial line indistinct; discal spots brown, separated; postmedial line black, obscure; cilia yellowish write. Hindwing pale gray, cilia grayish white.
Male genitalia (Fig. 3). Uncus oval, lateral margins enfolded at distal half. Apical process of gnathos conical, ca 1/3 length of uncus. Transtilla absent. Valva narrow, 5× as long as wide; clasper a narrowed triangular process, with a globular, haired base; costa stout, slightly longer than and ca 2/3 width of valva, its apex blunt, slightly forked; sacculus ca 2/5 length of valva, broader at base, tapering toward pointed apex, bearing dense, spine-like hairs along ventral margin. Juxta a broad, semicircular plate, with a pair of short, globular lateral lobes. Vinculum twice as long as its greatest width, narrowly rounded anteriorly. Aedeagus nearly as long as valva, slightly curved towards base, with a tuft of granulations near base; Cornuti two stout thorns, longer one slightly less than half length of aedeagus. Culcita one pair of long hair tufts, 2/3 length of valva.
Female genitalia (Fig. 5). Ovipositor triangular, 3× as long as wide. Apophyses posteriores slender, 3/4 length of apophyses anteriores. Eighth tergite 2/3 length of its width. Antrum strongly sclerotized, funnel-shaped, broader than eighth segment. Ductus bursae sclerotized, 1.2× as long as corpus bursae, of nearly equal width throughout, slightly broader at junction with corpus bursae. Corpus bursae heart-shaped, scobinate-granulate on inner surface, with two sclerotized patches: one oval sclerotized plate near middle; one irregular large plate from junction with ductus bursae to anterior 1/3, its posterior half smooth, forming a shallow fold along its edge, anterior half granulated and wrinkled. Ductus seminalis from posterior margin of corpus bursae. DNA barcode. One DNA barcode from a female paratype was obtained and deposited in GenBank (accession numbers: MT749678), DNA voucher slide no. DNAYLL18119.
Etymology. The specific name is derived from the Latin prefix tri-, meaning three, and the Latin word striatus, meaning streak, referring to three grayish black streaks on the forewing.
Distribution. China (Ningxia). Host plant. Unknown. ( Diagnosis. Adults (Fig. 2) with wingspan 15.0-22.0 mm. Pima boisduvaliella is characterized by the yellowish brown forewing with a white subcostal streak; the elongate valva with a well-developed costa that produced and weakly notched apically, the broad semicircular juxta with a pair of short, finger-like lateral lobes, the V-shaped vinculum ca 1.5× length of its greatest width, and the aedeagus with two thorns that slightly less than half the length of the aedeagus in the male genitalia (Fig. 4); the rounded antrum, the heart-shaped corpus bursae with dense microtrichia in anterior 1/3, with a small oval sclerotized plate and a couple of tortuous, sclerotized plates in the female genitalia (Fig. 6 (Fig. 7), Europe (Slamka 2019: 128, fig. 145), Canada, USA.

Discussion
Pima is a genus containing 25 species of which 15 are Palaearctic (Roesler 1990;Leraut 2014;Tsvetkov 2016;Vives Moreno and Gastón 2017;Slamka 2019), nine species are Nearctic (Heinrich 1956;Neunzig 2003), and two are Afrotropical (Joannis 1927) ( Table 1). Species of Pima might be expected to occur at higher elevations, as most of them were recorded from mountainous areas. In China (Fig. 7), P. boisduvaliella is mainly distributed in the north, but also occurred in the west, such as Xinjiang and Tibet; P. trifidella is distributed in Xinjiang; and P. tristriata sp. nov. is only found in Zhongwei, Ningxia. Adults of the two species were collected from mountain areas with altitudes ranging from 900 m to 3050 m.