Four new species of the primitively segmented spider genus Qiongthela from Hainan Island, China (Mesothelae, Liphistiidae)

Abstract The primitively segmented spider genus Qiongthela Xu & Kuntner, 2015 consists of seven species that are distributed in Hainan Island, China and southern Vietnam. Of the seven species, five are known from Hainan Island. In this study, four more Qiongthela species collected from Hainan Island are diagnosed and described as new to science based on morphological characters: Q. baotingsp. nov. (♂♀), Q. qiongzhongsp. nov. (♂♀), Q. sanyasp. nov. (♂♀), Q. yinggezuisp. nov. (♂♀). To facilitate future identification, the GenBank accession codes of the DNA barcode gene, cytochrome c oxidase subunit I (COI), for all the type specimens are also provided.

The genus Qiongthela was established by Xu and Kuntner in 2015 based on both morphological and molecular characters (Xu et al. 2015a, b). Until now, there are only seven named species, five of which are known from Hainan Island, China: Q. baishensis Xu, 2015, Q. bawang Xu, Liu, Kuntner & Li, 2017, Q. jianfeng Xu, Liu, Kuntner & Li, 2017, Q. wuzhi Xu, Liu, Kuntner & Li, 2017, and Q. yini Xu, Liu, Kuntner & Li, 2017; the other two of which, Q. australis (Ono, 2002) and Q. nui (Schwendinger & Ono, 2011), are distributed in southern Vietnam (Fig. 1C) (Ono 2002;Schwendinger and Ono 2011;Xu et al. 2015aXu et al. , b, 2017bWord Spider Catalog 2020). In this study, we diagnosed and described four more new Qiongthela species collected from Hainan Island based on both male and female genital morphology. In addition, we also provided the COI sequences of the holotypes for facilitating future identification.

Materials and methods
All specimens were collected from Hainan Island, China. All the type and voucher specimens are deposited at the College of Life Sciences, Hunan Normal University (HNU), Changsha, Hunan Province, China. We collected the spiders alive and fixed them in absolute ethanol if they were adults. For juvenile/subadult males, we took them back to the laboratory and reared them until they reached adulthood. We removed the right four legs of adults, preserved them in 100% ethanol and kept at -80°C for molecular work. We preserved the remains in 80% ethanol as vouchers for morphological identification and examination. We examined and dissected the specimens using an Olympus SZ51 stereomicroscope. We cleaned the female genitalia in 10 mg/ml trypsase (Bomei Biotech Company, Hefei, Anhui, China) for at least 3 hours at the room temperature to dissolve soft tissues. We took the photos under the Olympus BX53 compound microscope using a digital camera CCD, and generated compound focussed images using Helicon Focus v6.7.1. All measurements were carried out under a digital camera MC170HD mounted on stereomicroscope Leica M205C and given in millimeters. Leg and palp measurements are given in the following order: leg total length (femur + patella + tibia + metatarsus + tarsus), palp total length (femur + patella + tibia + tarsus).
Qiongthela baoting sp. nov. http://zoobank.org/C104261D-DBFB-4A70-84FD-5CF0BD15B82E Figure 2 Type material.  ductor with a pointed apex ( Fig. 2A-D); from all the other Qiongthela species by the contrategulum with four edges distally ( Fig. 2A, D), and by the marginal apophysis of the tegulum with a flake-like, semi-translucent apex ( Fig. 2A, D, F) . Female of Q. baoting sp. nov. differs from that of Q. baishensis and Q. nui by the base of the lateral receptacular clusters close to the inners, and by the genital stalks of the inners thicker than those of the laterals (Fig. 2G, H); from the other Qiongthela species by two paired receptacular clusters all along the anterior margin of the bursa copulatrix, with distinct genital stalks, and the inners larger than the laterals (Fig. 2G, H).
Palp. Cymbium with a short, thick projection dorsally (Fig. 2F); paracymbium unpigmented and unsclerotised prolaterally, with numerous setae at the tip ( Fig. 2A, B). Contrategulum with an irregular dentate edge proximally and four edges distally: the inner edge sharp, very short; the middle two edges serrate, one towards the proximal portion of contrategulum, the other ended at the centre of the contrategulum; the outer edge short, smooth, slightly sclerotised ( Fig. 2A, D, F). The marginal apophysis of tegulum long, wide basally, with a flake-like and semi-translucent apex distally ( Fig.  2A, D), a proximally directed terminal apophysis of tegulum with smooth margin, narrowing to a slightly hooked apex ( Fig. 2A-E). Conductor situated ventro-proximally on embolus, basal portion fused with embolus, distal free narrowing to a pointed apex ( Fig. 2A-E). Embolus largely sclerotised, with a wide, flat opening of sperm duct distally ( Fig. 2A, D, E).
Female (Fig. 1A). Carapace dark brown; opisthosoma reddish brown, with 12 redbrown tergites, close to each other, the first 2-7 larger than the others, and the fourth largest; sternum narrow, nearly twice as long as wide; a few fine pointed hairs running over the ocular area; chelicerae robust with promargin of cheliceral groove containing 10 denticles of variable size; legs with firm hairs and spines; 7 spinnerets. Measurements: BL 16.35, CL 7.30, CW 6.12, OL 7.59, OW 6.25; ALE > PLE > PME > AME; palp 10.09 ( Female genitalia. Two pairs of receptacular clusters along the anterior margin of the bursa copulatrix, close to each other, the inner ones distinctly larger than the laterals, with genital stalks thicker than those of the laterals (Fig. 2G, H).
Etymology. The species epithet, a noun in apposition, refers to the type locality. Diagnosis. Males of Q. qiongzhong sp. nov. resemble those of Q. yinggezui sp. nov., but can be distinguished from those of the latter by the marginal apophysis of the tegulum with a blunt apex (Fig. 3A, D); from Q. baoting sp. nov. by the tegulum marginal apophysis with a non-translucent apex (Fig. 3A, D), by the contrategulum with two edges distally (Fig. 3A, D), and by the cymbial projection long and thin (Fig.  3G); from Q. australis by the conductor with a slightly bent apex (Fig. 3C, E, G), and by the contrategulum lacking beak-like extension (Fig. 3F); from Q. jianfeng by the terminal apophysis of the tegulum abruptly narrowed distally ( Fig. 3A-C); from Q. nui, Q. sanya sp. nov., and Q. wuzhi by the marginal apophysis of the tegulum with a blunt apex (Fig. 3A, D). Females of Q. qiongzhong sp. nov. can be distinguished from those of Q. bawang and Q. jianfeng by the receptacular clusters with indistinct genital stalks (Fig. 3H, J); from those of Q. baishensis, Q. baoting sp. nov., Q. nui, Q. yini, and Q. wuzhi by the similar-sized receptacular clusters or the laterals slightly larger than the inners (Fig. 3H-M).
Description. Male (holotype). In alcohol carapace light reddish brown; opisthosoma light brown, with brown 12 tergites, close to each other, the first 2-7 larger than others, and the fourth largest; sternum narrow, nearly twice as long as wide; a few fine pointed hairs running over the ocular area; chelicerae with promargin of cheliceral groove containing 10 denticles of variable size; legs with firm hairs and spines; 7 spinnerets.  Palp. Cymbium with a long, thin projection dorsally (Fig. 3G); paracymbium unpigmented and unsclerotised prolaterally, with numerous setae at the tip (Fig. 3A, B). Contrategulum with a proximally irregular dentate edge and two distal edges: the inner one irregularly dentate, and the outer one sharp, semi-translucent (Fig. 3A, D-F). The marginal apophysis of tegulum with a blunt, slightly dentate apex distally, a proximally directed terminal apophysis of tegulum with several denticles and an abruptly narrowed and slightly hooked apex (Fig. 3A-E). Conductor situated ventro-proximally on embolus, the basal portion fused with embolus, distal free, narrowing to a slightly bent apex (Fig. 3A-C, E). Embolus largely sclerotised, retrolaterally with numerous longitudinal ribs, and with a wide, flat sperm duct opening distally (Fig. 3A, D, F).
Female genitalia. Two pairs of receptacular clusters along the anterior margin of the bursa copulatrix, receptacular clusters similar size or the inner ones slightly smaller than the lateral ones, with indistinct genital stalks (Fig. 3H-M).
Etymology. The species epithet, a noun in apposition, refers to the type locality.  Diagnosis. Males of Q. sanya sp. nov. can be distinguished from those of Q. baoting sp. nov. by the longer tegulum marginal apophysis with a non-translucent apex (Fig. 4A, D), and by the conductor with a bent apex (Fig. 4C, E, F, G); from those of the other Qiongthela species by the conductor base with a triangular apophysis ventrally (Fig. 4A-E). Females of Q. sanya sp. nov. can be distinguished from Q. australis, Q. yini and Q. yinggezui sp. nov. by the inner receptacular clusters smaller than the lateral ones ( Fig. 5A-H); from those of the other Qiongthela species by the inner receptacular clusters along the anterior margin of the bursa copulatrix, the laterals located slightly on the dorsal wall of the bursa copulatrix, and by the trapezoidal bursa copulatrix (Fig. 5A-H).
Palp. Cymbium with a short projection dorsally (Fig. 4G); prolateral side of paracymbium unpigmented and unsclerotised, with numerous setae at the tip ( Fig. 4A-C). Contrategulum with two distal edges: the inner one strongly dentate, and the outer one smooth, sharp, semi-translucent (Fig. 4A, D, F). Tegulum with a long, pointed, distally directed marginal apophysis, the proximally directed terminal apophysis with a dentate margin and continuously narrowing to a rounded, hooked apex ( Fig. 4A-E). Conductor situated ventro-proximally on embolus, fused with embolus at the basal portion, distal free narrowing to a bent apex (Fig. 4B, C, E-G); conductor base with a triangular apophysis ventrally (Fig. 4A-E). Embolus largely sclerotised, with a wide, flat sperm duct opening distally, retrolaterally with numerous longitudinal ribs (Fig. 4B, C, E).
Female genitalia. The inner receptacular clusters along the anterior margin of the bursa copulatrix, the lateral ones located slightly on the dorsal wall of the bursa copulatrix; the inner ones smaller than the lateral ones, with short or long genital stalks. The bursa copulatrix trapezoidal (Fig. 5A-H).
Variation. Males and females vary in body size. The range of measurements in males (N = 3): BL 13.40-15.01, CL 6.47-7.21, CW 5.87-6.53, OL 6.16-7.53, OW  . In addition, female genitalia show intraspecific variation: the inner pair of the receptacular clusters along the anterior margin of the bursa copulatrix upward, with short or long genital stalks ( Fig. 5A-E, G), or clusters toward the dorsal margin (Fig. 5F, H).
Etymology. The species epithet, a noun in apposition, refers to the type locality. Diagnosis. Male of Q. yinggezui sp. nov. differs from that of Q. australis by the conductor base wide and with a bent apex (Fig. 6A-F), and by the shorter paracymbium (Fig. 6A); from Q. nui by the embolus with a smooth surface retrolaterally (Fig. 6B, C, E); from Q. baoting sp. nov. by the cymbium with an elongated projection (Fig. 6F), and by the conductor with a bent apex (Fig. 6B-E); from Q. jianfeng, Q. qiongzhong sp. nov. and Q. sanya sp. nov. by the scutiform marginal apophysis of the tegulum thick basally and pointed distally ( Fig. 6A-F), and by the embolus with a smooth surface retrolaterally (Fig. 6B, C, E). Females of Q. yinggezui sp. nov. can be distinguished from those of Q. australis by the similar-sized receptacular clusters, and the lateral ones slightly located on the dorsal wall of the bursa copulatrix (Fig. 6G); from Q. yini by the receptacular clusters with more granula (Fig. 6G, H); from Q. sanya sp. nov. by the lack of genital stalks (Fig. 6G, H); from those of the other Qiongthela species by the inner receptacular clusters situated at the anterior margin of bursa copulatrix, the lateral pair located on the dorsal wall of the bursa copulatrix (Fig. 6G, H).
Female genitalia. Two paired of the similar-sized receptacular clusters, the inner ones along the anterior margin of the bursa copulatrix, and the lateral ones located slightly on the dorsal wall of the bursa copulatrix, without genital stalks (Fig. 6G, H).