A survey of Pireneitega from Tajikistan (Agelenidae, Coelotinae)

Abstract Five new species of Pireneitega species from Tajikistan are described: Pireneitega zonsteini sp. n. (♂♀), Pireneitega muratovi sp. n. (♀), Pireneitega tyurai sp. n. (♀), Pireneitega ramitensis sp. n. (♀) and Pireneitega kovblyuki sp. n. (♂). Pireneitega major (Kroneberg, 1875) is redescribed for the first time based on the lectotype designated here. DNA barcodes for the five new species are documented for future use and as proof of molecular differences between these species.


Introduction
Coelotinae is the largest subfamily of Agelenidae with more than 650 species distributed in the Holarctic and southeast Asia (World Spider Catalog 2016). Pireneitega Kishida, 1955 with 30 species distributed across the Palaearctic (World Spider Catalog 2016, Zhang et al. 2016) is one of the most species-rich genera of the subfamily. It is relatively well studied in comparison to other species-rich (and non-monophyletic) genera Coelotes Blackwall, 1841 and Draconarius Ovtchinnikov, 1999. The species of Pireneitega found in Caucasus and Xinjiang were recently revised (Kovblyuk et al. 2013;Zhang et al. 2016) but the genus remains poorly studied in Central Asia. Of three species known from central Asia (Mikhailov 2013: P. birulai (Ermolajev, 1927 (currently considered a junior synonym of P. luctuosa (L. Koch, 1878)), P. fedotovi (Charitonov, 1946) and P. major (Kroneberg, 1875)), P. fedotovi is known only from the original description and P. major only from two very short descriptions supplied with sketchy figures. A short trip by the junior author to Tajikistan revealed five new morphospecies of Pireneitega, each separated by distinct genetic gaps. The goal of this paper is to provide descriptions of the new species (including records of their molecular markers) and a redescription of P. major whose type locality lies in northern Tajikistan.

Material and methods
Specimens were examined and measured with a Leica M205C stereomicroscope. Images were captured with an Olympus C7070 wide zoom digital camera (7.1 megapixels) mounted on an Olympus SZX12 dissecting microscope. Epigynes and male palps were examined after dissection. Epigynes were cleared by boiling it in 10% KOH solution before taking photos of the dorsal view. All measurements are given in millimeters. Pireneitega major was photographed and drawn using an MBS-9 stereomicroscope with Pro-MicroScancamera. Leg measurements are given as: total length (femur, patella + tibia, metatarsus, tarsus).
Terminology used for copulatory organ characters in the text and figure legends follows Wang (2002) with some modifications.
Abbreviations used in the text and figure legends are: A epigynal atrium; ALE anterior lateral eye; AME anterior median eye; AME-ALE distance between AME and ALE; AME-AME distance between AME and AME; DNA barcodes were obtained for future use: a partial fragment of the mitochondrial gene cytochrome oxidase subunit I (COI) was amplified and sequenced for five new species using Primers LCO1490-oono (5'-CWACAAAYCATARRGATATTGG-3') (Folmer et al. 1994;Miller et al. 2010) and HCO2198-zz (5'-TAAACTTCCAGGT-GACCAAAAAATCA-3') (Folmer et al. 1994;. For additional information on extraction, amplification, and sequencing procedures, see Zhao et al. (2013). All sequences were blasted in GenBank; accession numbers are provided in Table 1.
Holotypes and some paratypes will be deposited in the Zoological Museum of the Moscow State University (ZMMU). Most paratypes are deposited in the Institute of Zoology, Chinese Academy of Sciences (IZCAS) in Beijing, China.

Note.
Pireneitega was long considered a nomen nudum (Yaginuma, in Brignoli 1983: p. 468). Kishida (1955), in a general survey of Agelenidae, considered Pireneitega to have been described by himself in 1928, although he had no publications that year. The genus "Pireneitega Kishida, 1928[Genotype: roscida (Koch, 1868]" was considered among the tribe Tegenariini Kishida, 1928(Kishida 1955). Although eye pattern was mentioned in the key to the genera of "Tegenariini", Kishida (1955) did not provide a formal description of the genus. Brignoli (1982) described Paracoelotes (type species Coelotes armeniacus Brignoli, 1978) from Turkey. Subsequently, Wang and Jäger (2007) revalidated Pireneitega with Paracoelotes as a junior synonym. Diagnosis. The chelicerae in most species of Pireneitega (including the type species) have 3 promarginal and 3 retromarginal teeth; other coelotines have either 2 or 4 retromarginal teeth (Zhang et al. 2016). The females can be distinguished by the widely separated epigynal teeth, the large atrium with subparallel margins, and the broad copulatory ducts ( Fig. 2A-B); other coelotines usually have a small atrium and copulatory ducts. The males can be distinguished by the absence of a dorsal "apophysis" on the conductor, the small RTA, and the distinct median apophysis ( Fig. 1A-C); other coelotines usually have a broad dorsal apophysis on the conductor and a reduced or indistinct median apophysis.
Composition. Thirty species of Pireneitega are known from Spain to Sakhalin (World Spider Catalog 2016; Mikhailov 2013). One species, P. major, was known from Tajikistan before the current study. Etymology. The species is named after Sergei Zonstein (University of Tel-Aviv, Israel) a partner of the junior author in the expedition to Tajikistan; noun (name) in genitive case. Diagnosis. The male can be distinguished from all other Pireneitega species except P. involuta (Wang et al., 1990) by having a broad conductor and thick patellar apophysis. From P. involuta it is distinguished by the blunt tip of the patellar apophysis (vs a tapering tip in P. involuta) ( Fig. 1; Wang et al. 1990: figs 13-15). The female can be distinguished from all other Pireneitega species except P. fedotovi by having a nearly trapezoidal atrium, long copulatory ducts, and short receptacles. From P. fedotovi it can be distinguished by its short epigynal teeth, about 0.5 times as long as length of the atrium (vs long epigynal teeth in P. fedotovi, about as long as the length of the atrium) (Fig. 2;Charitonov 1946: fig. 4).

Fe
Pt Ti Mt Ta   I  3d 2p 1r

Spination in male
Palp as in Fig. 1: patellar apophysis long, more than half length of tibia; tibia short, about 1/4 length of tarsus; VTA subequal to the tibial length, without pointed tip, extending beyond the tibia; RTA short, about 1/6 length of VTA; cymbial furrow long, more than half length of cymbium; conductor broad and with two spiraling loops; median apophysis broad and nearly triangular; embolus with broad base originating proximally on base of tegulum.
Epigyne as in Fig. 2A-B: epigynal teeth narrow and relatively short (shorter than width of atrium); septum short with weakly sclerotized tip, about 0.3 times as long as wide; atrium with well delimited posterior margin, about 1.3 times longer than wide, about 4 times longer than septum, subequal to width of septum; copulatory opening hidden by anterior margin of atrium; receptacles long, about 2 times longer than wide, separated by 2.5 times their diameters; copulatory ducts with 3 parts, the basal part running from receptacle posteriorly (Bd), median part running anteriorly (Md), and terminal part (Td) running posteriorly and leading to copulatory opening; median part as wide as terminal and 2 times longer than basal part; median part 1.5 times longer than receptacle; median parts touching each other; hoods indistinct.
Spination in female

Distribution.
Known only from the type locality (Fig. 8). Etymology. The species is named after Tajikistan zoologist Rustam Muratov (Dushanbe, Tajikistan) who was very helpful in organizing the expedition to Tajikistan; noun (name) in genitive case.
Diagnosis. The female can be distinguished from all other Pireneitega species except P. fedotovi, P. luniformis (Zhu & Wang, 1994), and P. major by having narrow epigynal teeth and an elongate oval atrium. It can be distinguished from P. fedotovi by the pointed tip of septum (vs blunt tip in P. fedotovi), from P. luniformis by the elongate oval receptacles (vs spiralled in P. luniformis), and from P. major by its short epigynal teeth, ca. 0.8 times as long as length of the atrium (vs long epigynal teeth in P. major, about as long as the length of the atrium) (Figs 3, 7;Charitonov 1946: fig. 4;Zhu and Wang 1994: figs 5-6).
Epigyne as in Fig. 3A-B: epigynal teeth narrow, their length equal to width of the narrowest part of the atrium; septum with well delimited tip, ca. 0.5 times as long as wide; copulatory opening distinct; atrium with well delimited posterior margin, about 1.4 times longer than wide, ca. 2 times longer than and 0.7 times as wide as septum; receptacles long, about 2.5 times as long as wide, bases of receptacles separated by 2 diameters; copulatory ducts with 3 parts, median part as long as receptacles, and anterior part slightly wider than receptacles; hoods indistinct. Spination Distribution. Known only from the type locality (Fig. 8). Etymology. The species is named after Sergei V. Tyura (Magadan, Russia) a friend of the junior author; noun (name) in genitive case.
Diagnosis. The female can be distinguished from all other Pireneitega species except P. tianchiensis (Wang et al., 1990) by having short receptacles and copulatory ducts. It can be distinguished from P. tianchiensis by the broad and long epigynal teeth, about 0.85 times as long as atrium (vs short and narrow epigynal teeth in P. tianchiensis, about 0.5 times as long as atrium) (Fig. 4A-B; Wang et al. 1990: figs 84-85).
Epigyne as in Fig. 4A-B: epigynal teeth long (nearly as long as atrium); septum with weakly sclerotized tip, about 0.5 times as long as wide; atrium with weakly sclerotized posterior margin, about 0.7 times as long as wide, about 1.8 times longer than and 0.7 times as wide as septum; copulatory opening hidden; receptacles large, ca. 2 times longer than wide; copulatory ducts with two parts, terminal parts (Tp) not touching each other, about 0.5 length of receptacles, basal parts (Bp) shorter than width of receptacle; hoods indistinct. Spination Distribution. Known only from the type locality (Fig. 8).   Etymology. The specific name is an adjective and refers to the type locality; adjective. Diagnosis. The female can be distinguished from all other Pireneitega species except P. muratovi sp. n., P. fedotovi, P. luniformis and P. major, by having an elongate oval atrium, narrow epigynal teeth, and long copulatory ducts. It can be distinguished from P. muratovi sp. n. and P. luniformis by the narrow tip of the copulatory ducts (vs round tip in P. muratovi sp. n. and P. luniformis) and from P. fedotovi and P. major by the bent epigynal teeth (vs straight epigynal teeth in P. fedotovi and P. major) (Figs 3, 5, 7;Charitonov 1946: fig. 4;Zhu & Wang 1994: figs 5-6).
Palp as in Fig. 6A-C: patellar apophysis absent; tibia long, ca. 0.5 length of cymbium; VTA short and wide, about 1/3 length of tibia, without pointed tip; RTA short, about 1/5 length of VTA, poorly visible; cymbium long, its tip as long as or longer than genital bulb; conductor short, with hook-shaped, partially looped tip, tip located distally from tegulum; median apophysis broad and nearly triangular; embolus with broad, nearly tongue-shaped base, beginning at 6:30 o'clock position. Spination  Schenkel (1936), Hu and Wu (1989), and Song et al. (1999;copied from Hu and Wu 1989) are of a species other than P. major, the identity of which is currently unknown. All records of this unknown species are from Xinjiang, China.
Diagnosis. This species is easily distinguished from other species of Pireneitega found in Tajikistan by its larger size (carapace length >6 mm vs <4.75) and having 5 spines on tarsus IV (vs other species with 0-4). The epigyne of P. major is most similar  to that of P. muratovi sp. n. and P. ramitensis sp. n. It can be distinguished from P. muratovi sp. n. by its shorter receptacles with length/width ratio of 2.3 (vs 2.6 in P. muratovi), shape of copulatory ducts, and shorter teeth (cf. Figs 3A-B and 7A-B). Pireneitega major can be separated from P. ramitensis sp. n by its wider epigynal atrium and shorter, wider receptacles as well as by its shorter and wider copulatory ducts (cf. Figs 5A-B and 7A-B).