Asianopis gen. nov., a new genus of the spider family Deinopidae from Asia

Abstract A new genus of the spider family Deinopidae C.L. Koch, 1850 is described from Asia: Asianopis Lin & Li gen. nov., with A. zhuanghaoyuni Lin & Li sp. nov. as the type species. The new genus is divided into two species groups, of which the liukuensis-group includes two species: A. dumogae (Merian, 1911) sp. reval. comb. nov. (♀) and A. liukuensis (Yin, Griswold & Yan, 2002) comb. nov. (♂♀); and the zhuanghaoyuni-group comprises five species: A. celebensis (Merian, 1911) comb. nov. (♂), A. konplong (Logunov, 2018) comb. nov. (♂), A. wangi Lin & Li sp. nov. (♂♀), A. wuchaoi Lin & Li sp. nov. (♂♀), and A. zhuanghaoyuni Lin & Li sp. nov. All previously described species are transferred from Deinopis MacLeay, 1839. Deinopis scrubjunglei Caleb & Mathai, 2014 is treated as a junior synonym of Asianopis liukuensiscomb. nov.


Introduction
The spider family Deinopidae C.L. Koch, 1850 (Araneae, Deinopoidea), known as net-casting or ogre-faced spiders, is a small family that consisted of two genera and 64 species prior to the current study (World Spider Catalog 2019). The genus Deinopis was established by MacLeay (1839) based on Deinopis lamia MacLeay, 1839 (♂♀) from Cuba. The other genus, Menneus, was established by Simon (1876) based on Menneus tetragnathoides Simon, 1876 (♂) from Angola.

Material and methods
All specimens were preserved in 80% ethanol. Metatarsi and tarsi were removed for preservation in 100% ethanol for subsequent molecular work. Epigynes were cleared in proteinase K at 56 °C to dissolve non-chitinous tissues for three hours. Specimens were examined under a LEICA M205C stereomicroscope. Photomicroscope images were taken with an Olympus C7070 zoom digital camera (7.1 megapixels). Laboratory habitus photographs were taken with a Canon 5D Mark III digital camera equipped with a Canon MP-E 65 mm lens. Photos were stacked with Helicon Focus (version 6.7.1) or Zerene Stacker (version 1.04) and processed in Adobe Photoshop CC 2018. Photographs of Asianopis celebensis comb. nov. were taken by a KEYENCE. Photographs of Asianopis liukuensis comb. nov. from India (i.e., the type materials of D. scrubjunglei) were taken using a Leica DFC500 HD camera mounted on a Leica M205A stereomicroscope.
All measurements are in millimetres. Eye sizes are measured as the maximum diameter from either the dorsal or frontal view. Leg measurements are given as follows: total length (femur, patella+tibia, metatarsus, tarsus). Copulatory duct turns are defined by the number of apparent loops on the lateral margin of the copulatory/fertilization duct complex in dorsal view. The length of the embolic tip fold is measured as from the beginning of the fold to the embolic tip (Fig. 22D, E). The terminology used in the text and figures follows Coddington et al. (2012). Distribution maps were generated using ArcMap software (version 10.2).
A total of 31 specimens of Deinopidae were collected for phylogenetic analysis (Suppl. material 1: Table S1). Sequences of seven specimens were from the National Center for Biotechnology Information (NCBI) public data, and the other 24 were from recent field collections. Whole genomic DNA was extracted from 2-4 legs using a TIANamp Genomic DNA kit (TIANGEN Inc., Beijing, China) following the manufacturer's protocol. Seven gene fragments were amplified in 20-μL reactions: COI (~640 bp), 12S (~330 bp), 16S (~470 bp), 18S (~1700 bp), 28S (~1200 bp), H3 (~310 bp) and wnt (~330 bp). Primers and PCR conditions for each locus are listed in Suppl. material 1: Table S2. Sequence chromatograms were proofed and edited using Sequencher version 4.2 Demo (Gene Codes Corporation, Ann Arbor, MI USA). The COI, H3 and wnt fragments were translated in MEGA version 7 (Kumar et al. 2016) to check for the presence of stop codons. A representative of the family Uloboridae was used as the outgroup, with the corresponding sequences downloaded from NCBI. The complete list of 32 taxa and GenBank accession numbers are provided in Suppl. material 1: Table S1.
Multiple sequence alignments were carried out with MAFFT version 7.243 (Katoh and Standley 2013). Alignments of the protein-coding COI, H3 and wnt genes were produced using the L-INS-i method. As for the highly variable ribosomal genes, the E-INS-i method was used to generate alignments of 12S, 16S, 18S, and 28S. To exclude the ambiguously aligned regions, alignments of the ribosomal genes were processed with the program trimAl version 1.3 (Capella-Gutiérrez et al. 2009). The alignments are shown in the supplementary data.
The concatenated gene matrix was partitioned by gene using PartitionFinder version 1.1.1 (Lanfear et al. 2012). The best partitioning scheme was selected based on the Akaike information criterion (AIC) (Suppl. material 1: Table S3). Maximum likelihood (ML) analysis was performed using RAxML version 8.2.9 with a GTR + Γ + I model applied to each partition (Stamatakis 2014). One thousand non-parametric bootstrap replicates were conducted to obtain the best-scoring ML tree.
Bayesian analysis was performed using MrBayes version 3.2.6 (Ronquist et al. 2012). Two independent runs, each with four independent chains, were carried out for 20,000,000 generations and were sampled every 1,000 generations with a burn-in of 25%. Partitions and models followed the result of PartitionFinder. Convergence of the runs was determined with the standard deviation of split frequencies (<0.01). Effective sampling sizes (>200) of all parameters were checked in Tracer version 1.6 (Rambaut et al. 2014). A 50% majority-rule consensus tree was then constructed from the postburnin sampled trees to estimate posterior probabilities (PP). Etymology. The generic name is a combination of the word "Asia", referring to the distribution of the genus, and the generic name Deinopis. The gender is feminine.

Abbreviations
Diagnosis. Asianopis gen. nov. can be easily distinguished from Deinopis by the following characters: a prominent setal fringe can be found above the posterior median eyes in both sexes of Asianopis species (Fig. 4A, B), which is absent in Deinopis (Coddington et al. 2012: fig. 3a); the embolic tip of male Asianopis has an embolic middle apophysis (liukuensis-group, Fig. 21A), an embolic terminal apophysis or is weakly folded apically (zhuanghaoyuni-group, Fig. 21B-E), whereas none of these characters is present in Deinopis (Coddington et al. 2012: fig. 11m); the MADL in Asianopis is small and has a basal lobe, while in Deinopis, the median apophysis is larger than the MABL and covers the entire base (Coddington et al. 2012: fig. 11m); female chelicerae with many denticles between the promarginal and retromarginal teeth ( Description. Male. Total length 12.14-16.10 (n = 8), carapace pear-shaped, yellow-brown (liukuensis-group) or brown (zhuanghaoyuni-group) with white edge, white line extending from cephalic area to posterior margin and small spines sparsely dis-tributed; fovea longitudinal, indistinct. Chelicerae with a promarginal tooth and one or two retromarginal teeth (liukuensis-group) or with four promarginal teeth and 2-6 retromarginal teeth (zhuanghaoyuni-group), no denticles. Endites and labium brown, distally white; sternum diamond-shaped, brown with median light band and few small spines. Legs brown, ventrally with black pattern and short spines, leg formula 1243. Opisthosoma cylindrical, brown or dark-brown with small black spots and irregular pattern. Cribellum entire, spinnerets brown (Figs 4,10,13,16).
Epigyne with anchor-shaped median plate, CO distinct, CD with three turns, S oval, SpD consistently wide (liukuensis-group) or with a well-developed MP, obscuring CO, CD with 7-8 turns, S oval, SpD consistently thin (zhuanghaoyuni-group).
Molecular phylogeny. The molecular phylogenetic analysis indicates with strong support that all the species in this study do not belong to Deinopis. Based on the 4893 bp-aligned sequences of seven gene fragments, the ML and Bayesian analyses produced the same topology, showing a split of a Southwest China clade from other clades and is strongly supported (Bootstrap value: 88; PP: 0.98) ( Natural habitat. All the species of Asianopis gen. nov. were collected from bushes in low-elevation forests. Composition. This new genus comprises two species groups: the liukuensis-group with two species: A. dumogae (Merian, 1911) sp. reval. comb. nov. and A. liukuensis (Yin, Griswold & Yan, 2002) comb. nov. and the zhuanghaoyuni-group with five species: A. celebensis (Merian, 1911)
Diagnosis. This species can be distinguished from A. liukuensis comb. nov. by the MP nearly covering the CO, S round, and the overall equal thickness of the CD (Figs 4, 6).
Description. See Merian (1911). Photos of the epigyne of the syntype are given in Figure 6.
Distribution. Indonesia (North Sulawesi). Comments. Merian (1911) reported D. celebensis based on three specimens from different localities in Sulawesi, Indonesia. One male (NMB-ARAN-00514b, "Zen-tral-Celebes, nördlich vom Golf von Bone", South Sulawesi, north of the Gulf of Boni (precise locality not known), one female from North Sulawesi (NMB-ARAN-00514a, "Wald bei Duluduo", Sulawesi Utara, forest near Duluduo, 00°31'33"N, 123°57'10"E and one female from Central Sulawesi (NMB-ARAN-00514c, Larga, südlich vom Posso-See, unterhalb Patiro Rano, bei 900 m, Central Sulawesi, south of Lake Poso at an elevation of 900 m (the localities "Larga" and "Patiro Rano" could not be located on maps; the epigyne of this specimen is missing, but the specimen is clearly larger than the others).   Merian (1911) stated that the male and the females may not represent the same species and suggested the name D. celebensis for the male, and D. dumogae for the female. According to the International Code of Zoological Nomenclature (International Commission on Zoological Nomenclature 1999: Article 11.5.1), such conditionally proposed species names are potentially available as valid names if published before 1961. The species has not been listed in any of the catalogues. We examined the types and concluded the male and the two females are indeed three different species. The palp of the male D. celebensis exhibits features of the zhuanghaoyuni group: the tegulum is obscured by the embolic coil, and the embolus is long and strongly coiled around the MA. The female from North Sulawesi (Doloduo) has features of the liukuensis group: an anchor-shaped median plate, CO distinct, CD with three turns. Thus, we revalidated the female D. dumogae as Asianopis dumogae (Merian, 1911), sp. reval. comb. nov.    Diagnosis. This species can be distinguished from other congeners by the distinct female copulatory opening, oval S, and CD tapering from the copulatory opening to spermatheca (Figs 6, 8).
Comments. Type materials of D. scrubjunglei syn. nov. were examined and no differences between A. liukuensis and D. scrubjunglei were observed. Thus, we consider D. scrubjunglei to be a synonym of A. liukuensis, and the figures of D. scrubjunglei are given for comparison (Figs 7, 8, 19C).
Diagnosis. The male can be distinguished from other congeners by having the distal lobe of the MA distinctly smaller than the basal lobe; in other Asianopis spp., the distal lobe is slightly smaller than the basal lobe (Fig. 9A, C).
Description. See Merian (1911). Photos of holotype male habitus and palps are shown in Fig. 9A-F.
Etymology. The species is named after Mr Dongdong Wang, the collector of the holotype; noun (name) in genitive case.
Diagnosis. The males resemble A. zhuanghaoyuni sp. nov. but can be distinguished from other species by the ratio of the length of the embolic opening to the length of the embolic tip fold, which is 1:6 in A. wangi sp. nov. and 1:8 in A. zhuanghaoyuni sp. nov. The fold is more developed in A. wangi sp. nov. (Fig. 21C, D). The median plate is triangular in A. wangi sp. nov. and subtriangular in A. zhuanghaoyuni sp. nov. (Figs 12, 19).
Distribution. China (Hainan).  Etymology. The species is named after Mr Chao Wu, the collector of the holotype male; noun (name) in genitive case.