Caucasiana 3: 119-126 (2024) DOI: 10.3897/caucasiana.3.e120656 =, CAUCASIANA Journal on the biodiversity of the Caucasus and the adjacent regions Research Article A new Psammitis species (Araneae, Thomisidae) from an extinct volcano in Georgia with reevaluation of the generic position of Xysticus marmoratus Thorell, 1875 Armen Seropian'’®, Levan Mumladze2® 1 Institute of Ecology, Ilia State University, Cholokashvili av. 3/5 Tbilisi, 0162, Georgia 2 Institute of Zoology, Ilia State University, Cholokashvili av. 3/5 Tbilisi, 0162, Georgia Corresponding author: Armen Seropian (armen. seropiani@iliauni.edu.ge) OPEN Qrceess Academic editor: Nils Hein Received: 9 February 2024 Accepted: 29 March 2024 Published: 29 April 2024 ZooBank: https://zoobank. org/21D13F14-2F8D-41BE-BD32- 80900D3C3628 Citation: Seropian A, Mumladze L (2024) A new Psammitis species (Araneae, Thomisidae) from an extinct volcano in Georgia with reevaluation of the generic position of Xysticus marmoratus Thorell, 1875. Caucasiana 3: 119-126. https://doi.org/10.3897/ caucasiana.3.e120656 Copyright: © Seropian and Mumladze This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0). Abstract Anew species, Psammitis abuliensis sp. nov., from Didi Abuli Mt. (Georgia, Samtskhe-Ja- vakheti region), is described based on two male specimens. Diagnostic drawings, mea- surements, collecting data, diagnosis, and DNA barcodes are given. A new combination, Psammitis marmoratus (Thorell, 1875), comb. nov. (ex. Xysticus), is proposed based on the COI subunit barcode results and morphological characters. Key words: Arachnida, Caucasus, crab spider, description, diagnosis, new taxa, Samtskhe-Javakheti Introduction The long and complicated taxonomic history of Psammitis Menge, 1876, in- volves its separation from Xysticus C.L. Koch, 1835, and recognition as an in- dependent genera (Menge 1876; Dalmas 1922; Ono 1978; Wunderlich 1987; Lehtinen 2002; Breitling 2019), alternatively with synonymizations with Xysti- cus (Simon 1895; Ono 1988). To date, 32 taxa (30 species and 2 subspecies) of Psammitis are known, mostly distributed in the Palaearctic Region (WSC 2023), characterized by different diameters of lateral eyes and palp with anapophys- ate disc-shaped flat tegulum at best having simple folds or sclerotized ridges. From the seven species reported from the Western Palaearctic (Nentwig et al. 2023), only two congeners ~ P. ninnii (Thorell, 1872) and P. sabulosus (Hahn, 1832) - are recorded in the Caucasus and Georgia (Otto 2023; WSC 2023). In September 2018, two male specimens of an unknown Psammitis species were sampled by the second author on Didi Abuli Mt. - an extinct volcano and the highest peak in the Lesser Caucasus in Georgia, located in the Abul-Samsa- ri Range. Herein the specimens are described as Psammitis abuliensis sp. nov., accompanied by photos of the holotype, paratype, and terra typica, diagnostic drawings, collecting details, diagnosis, barcoding results, and a discussion on the generic placement of Xysticus marmoratus Thorell, 1875, based on the phy- logenetic analysis. 119 Seropian and Mumladze: A new species of Psammitis (Araneae, Thomisidae) from South-West Georgia Materials and methods The material for the present study was collected on September 12, 2018, during fieldwork in Javakheti Highland, at the peak of the mountain Didi Abuli, by hand under small stones. The collected material was preserved in 96% ethanol and stored in a freezer under -22°C at the scientific collections of Ilia State University (ISU), Georgia, Tbilisi, for further DNA barcoding. Photos of the preserved speci- mens (Figs 1-3) were taken using a Canon EOS 60D camera with a Canon EF-S 60mm f/2.8 Macro USM and Raynox DCR-250 Super Macro Snap-On Lens and a Canon Macro Twin Lite MT-26EX-RT attached, and the whole set was mounted on a Novoflex Castel-L Focusing Rack. Digital images were prepared using Zerene Stacker image stacking software and Adobe Photoshop CS6 (version 13.0). Diag- nostic drawings were made based on microscope photographs using a Wacom CTH-690 Intuos Medium Pen and Touch Tablet with the programs Krita (version 2.9.7) and Photoshop CS6 (version 13.0). All measurements are given in mm. The following abbreviations are used in the text, tables, and figures: ALE (an- terior lateral eyes), AME (anterior median eyes), d (dorsal), Et (embolus tip), Fe (femur), Mt (metatarsus), Pa (patella), p (prolateral), PLE (posterior later- al eyes), PME (posterior median eyes), r (retrolateral), Ta (tarsus), Ti (tibia), TuA (tutacular apophysis), v (ventral). Leg spination system according to Ono (1988). The prolateral femoral spines of leg | and ventral metatarsal spines of leg Il are not paired, thus the number is indicated. DNA processing DNA extraction at the ZFMK (Zoological Research Museum Alexander Koenig) followed the standard protocols of the GBOL (German Barcode of Life) project (Geiger et al. 2016; http://www.bolgermany.de). DNA extraction at ISU was per- formed following the implemented protocol (Seropian et al. 2023a). Extracted DNA was deposited in the scientific collections of Ilia State University, Tbilisi, Georgia, and aliquots will be deposited at The Leibniz Institute for the Analysis of Biodiversity Change (LIB) Biobank at Museum Koenig, Bonn, Germany, while the sequences have been submitted to Barcode of Life Data System (BOLD) databases (http://www.boldsystems.org/index.php). The newly obtained DNA barcodes of COI sequences were checked against the BOLD Systems data- base. The Barcode Index Numbers (BIN) (Ratnasingham and Hebert 2013) for the sequenced taxa and their nearest neighbor in BOLD Systems (if they had a BIN) are also given. For the calculation of sequence differentiation, we used p-distance as performed in the BOLD Systems. In addition to the newly generated DNA barcodes, the published CO! subunit barcodes of Psammitis spp. deposited in BOLD Systems and NCBI GenBank, along with a single barcode of P. ninnii originating from Georgia (obtained from the specimen collected within the CaBOL project), were included to evaluate relationships between Psammitis spp. based on uncorrected p-distance. As outgroups, COI barcodes obtained from the locally collected Xysticus spp., Spir- acme spp., and Bassaniodes spp. were used (Table 1). A Neighbor-Joining tree based on pairwise distances with 1000 bootstrap replicates was constructed with MEGA 11 software (Tamura et al. 2021) to visualize the phylogenetic relationships among specimens (Fig. 8). Caucasiana 3: 119-126 (2024), DOI: 10.3897/caucasiana.3.e120656 120 Seropian and Mumladze: A new species of Psammitis (Araneae, Thomisidae) from South-West Georgia Table 1. List of the CO] sequences used for molecular analyses. Species Country of origin | CaBOL-ID number | BOLD/GenBank Acc. N. Psammitis abuliensis sp. nov. 1012780 BOLD:AFP6894 Psammitis abuliensis sp. nov. 1012781 BOLD:AFP6894 Psammitis ninnii Georgia 1012634 BOLD:ACU8868 Spiracme Striatipes BOLD:AAD6911 Xysticus acerbus Georgia 1010349 BOLD:AAG1214 Xysticus marmoratus BOLD:AAF8321 Bassaniodes pseudorectilineus* Georgia 1010063 BOLD:AAO1 746 Psammitis deichmanni Serge _|__ tothe _. BOLD:AAB7094 Psammitis labradorensis Canada BOLD:AAB1154 Psammitis rugosus ——— BOLD:AE01986 Psammitis sabulosus Norway BOLD:ABU5838 Psammitis sabulosus eee BOLD:ABU5838 Psammitis ninnii | Spain fs M9985889.1 * The first record in Georgia, which will be featured in an upcoming article. Results Family Thomisidae Sundevall, 1833 Genus Psammitis Menge, 1876 Type species. Thomisus sabulosus Hahn, 1832 Psammitis abuliensis sp. nov. FIgs:1=5 https://zoobank.org/325DC4FB-4400-4DE1-8803-B7FODA12527B Type material. Holotype: 4 (CaBOL-ID 1012781): Georgia: Samtskhe-Javakheti, Akhalkalaki municipality, Didi Abuli Mt.; N41.4380°, E43.6462°; 3267 m a.s.l., under rocks; 12 September 2018; leg. L. Mumladze. Paratype: 1’ (CaBOL-ID 1012780): same collecting data. Both specimens are deposited in the scientific collections of Ilia State University, Georgia, Tbilisi. Diagnosis. The new species resembles P. sabulosus (Hahn, 1832) that occurs in Georgia (Thaler 1981: figs 52, 57; Jantscher 2002: figs 5-6) and P. demirsoyi (Demir, Topcu & Tiirkes, 2006) from Turkey (Demir et al. 2006: figs 1-2, 5-7). The males of the new species can be distinguished from those of P. sabulosus by having a bow tie-shaped very short and broad embolus tip (vs. long and thin), a longer basal embolus, and a larger tutacular apophysis. From the males of P. demirsoyi, those of P. abuliensis sp. n. can be distinguished by the presence of a large tutacular apophysis (vs. absence) and a bow tie-shaped very short and broad embolus tip (vs. differently shaped longer and thinner) (Figs 3, 5). Description. Male (holotype/paratype). Total length: 6.90/6.85; Carapace: 3.41/3.39 long, 3.35/3.35 wide; Abdomen: 3.80/3.85 long, 3.30/3.32 wide; Clypeal height: 0.35/0.34; Clypeus with 15 long and robust bristles; Chelicer- al length: 1.15/1.15; AME: 0.11/0.11; ALE: 0.20/0.20; PME: 0.11/0.11; PLE: 0.14/0.14. Color and pattern as shown in Figs 4-5. Sternum yellow, with irreg- Caucasiana 3: 119-126 (2024), DOI: 10.3897/caucasiana.3.e120656 121 Seropian and Mumladze: A new species of Psammitis (Araneae, Thomisidae) from South-West Georgia Figures 1-5. Psammitis abuliensis sp. nov., male (1 holotype, dorsal habitus 2 paratype, dorsal habitus 3 left palp, ventral view 4 ditto, retrolateral view 5 tip of embolus, rotated). Scale bars: 2 mm (1-2); 0.2 mm (3-4); 0.1 mm (5). ular dark spots. Leg coloration: Fe and Pa I-IV dark brown (Fe and Pa III-IV apically lightened), Ti, Mt, and Ta I-IV dark yellow (Ti I-II basally darkened). Leg measurements and spination as in Tables 2 and 3. Female unknown. Etymology. The specific epithet is an adjective referring to the type locality of the new species in Akhalkalaki municipality. Habitat. Two males of the new species were obtained by hand collecting under the small stones on a mountaintop (Figs 6-7). Distribution. Known from the type locality only. Barcoding results. Two identical barcodes were obtained from the specimens with CaBOL-IDs 1012780 and 1012781 (BOLD:AFP6894) with the nearest neighbor inthe BOLD Systems P. ninnii from Spain with a private status (p-distance 6.3%). Table 2. Length (mm) of leg segments. Male (holotype/paratype). Tota | 6.517351 1.42/1.41 2.60/2.58 2.50/2.50 1.21/1.20 11.24/11.20 II 3.31/3.30 Tee 2h leoz: 2:52/2:50 211/214 M21 2A 10.67/10.64 III T5S/1.53 1 ht Al 2.04/2.02 MAZZA 1.04/1.04 7.46/7.41 IV 22/222 1.02/0.98 2.11/2.09 1.80/1.80 10271202 8.20/8.11 Caucasiana 3: 119-126 (2024), DOI: 10.3897/caucasiana.3.e120656 122 Seropian and Mumladze: A new species of Psammitis (Araneae, Thomisidae) from South-West Georgia Table 3. Leg spination. ee ae me d0-1-1-1-1 p11 pi=t-161-1-1 p1-1-1-1-lap r1-1-1-2ap v2-1-2-2-2-1-2ap v1-1-2-1-1-lap ddepe2- 1-4 past leel=121 p1-1-1-lap r1-1-1ap V2-1= 11-11 lZap v11-2ap dO-1-1-1-1-1 p1-1-lap r1-1-0 p1-1-lap r1-1-lap sel lat Pa v2-2-2 d1-1-1 4 Re id FL p1-1-lap r1-1-1lap V1-1-1-1-2ap v1-1-1-1-2ap Figures 6-7. Type locality of Psammitis abuliensis sp. nov. in Georgia, Didi Abuli Mt. (6 — sampling area; 7 - sampling plot). Psammitis marmoratus (Thorell, 1875), comb. nov. Xysticus marmoratus Mcheidze, 1997: 163, figs 305-306 (3). Xysticus embriki Hepner et al., 2011: 38, figs 5-10 (4). Note. For full nomenclatural references see WSC (2023). Comments. This species is transferred to this genus due to the similarity of the male palp to the generotype, which possesses an anapophysate tegulum. As for the Psammitis spp. females, there seem to be no defined characters allowing reliable separation from Xysticus spp. The generic reevaluation is also supported by the phylogenetic tree generated within the preset study, which includes Xys- ticus marmoratus, a species not involved in a subset of the analyses performed by Breitling (2019), who supported the recognition of Psammitis as an indepen- dent genus. Our results suggest a close relationship between X. marmoratus and Psammitis spp. (Fig. 8); therefore, we propose a new combination, Psammitis marmoratus (Thorell, 1875), comb. nov. (= Xysticus marmoratus Thorell, 1875). Discussion Recent studies and collecting efforts in Georgia within the last half a decade have resulted in an additional 5 species of Thomisidae, of which one species is new to science (the herein-described Psammitis abuliensis sp. nov.) (Seropian Caucasiana 3: 119-126 (2024), DOI: 10.3897/caucasiana.3.e120656 123 Seropian and Mumladze: A new species of Psammitis (Araneae, Thomisidae) from South-West Georgia 39 90 HH 0.010 68 68 CaBOL-1012780|Psammitis abuliensis. Georgia|COI-5P 100 CaBOL-1012781|Psammitis abuliensis. Georgia|COI-5P CaBOL-1012634||Psammitis ninnii. Georgia|COI-5P| MW398589.1||Psammitis ninnii. Spain|COI-5P| YDBB1043-21|Psammitis rugosus. Canada|COI-5P ACHAR1773-18|Psammitis deichmanni. Canada|COI-5P|MN675458 GBA23879-15|Psammitis labradorensis. Canada|COI-5P|KJ496494 CaBOL-1016820]||Xysticus marmoratus. Georgia|COI-5P| ARTRD244-15|Psammitis sabulosus. Norway|COI-5P 100 FBARB567-11|Psammitis sabulosus. Germany|COI-5P|KX537376 CaBOL-1010063||Bassaniodes pseudorectilineus. Georgia|COI-5P| CaBOL-1010349)||Xysticus acerbus. Georgia|COI-5P| CaBOL-1023862||Spiracme striatipes. Georgia|COI-5P| Figure 8. Phylogenetic relationships are presented by the Neighbor-Joining tree based on the mitochondrial COI barcode using the p-distance model with other default parameters provided by Mega 11. The analyses involved 13 COI nucleotide sequences of 10 thomisid species. Numbers indicate bootstrap support values from 1000 replicates. et al. 2023a, b; present article). The herein reevaluation of P. marmoratus (ex. Xysticus) and the result of the research conducted by Breitling (2019) suggest a strong need for further revision of Xysticus spp. The exploration of remote, poorly studied, and hard-to-reach localities in Georgia could result in findings of more new species and redescriptions of very poorly defined three species en- demic to the Caucasus region, namely X. caucasius L. Koch, 1878, X. charitono- wi Mcheidze, 1971, and X. kalandadzei Mcheidze & Utotschkin, 1971, described by single females and known from single publication(s). Acknowledgements AS is indebted to Nino Kachlishvili for performing the DNA extraction from the specimens. We are grateful to Nils Hein (subject editor), the anonymous re- viewer, and Konrad Wisniewski for providing constructive and valuable sugges- tions on the manuscript. The responsibility for the content of this publication lies with the authors Caucasiana 3: 119-126 (2024), DOI: 10.3897/caucasiana.3.e120656 124 Seropian and Mumladze: A new species of Psammitis (Araneae, Thomisidae) from South-West Georgia Additional information Conflict of interest The authors have declared that no competing interests exist. Ethical statement No ethical statement was reported. Funding The fieldwork of LM was supported by the Conchological Society of Great Brit- ain & Ireland under the small research grant framework (https://conchsoc.org/ pages/grants.php). DNA research was founded by the Federal Ministry of Edu- cation and Research under grant number 01DK20014A. Author contributions AS made a morphological description of the specimens and wrote the first draft of the manuscript. LM Collected material and revised the final version of the manuscript. Author ORCIDs Armen Seropian ® https://orcid.org/0000-0003-3777-9954 Levan Mumladze © https://orcid.org/0000-0002-2172-6973 Data availability All of the data that support the findings of this study are available in the main text or Supplementary Information. References Breitling R (2019) A barcode-based phylogenetic scaffold for Xysticus and its relatives (Araneae: Thomisidae: Coriarachnini). Ecologica Montenegrina 20: 198—206. https:// doi.org/10.37828/em.2019.20.16 Dalmas R (1922) Catalogue des araignées récoltées par le Marquis G. Doria dans l'ile Gi- glio (Archipel toscan). Annali del Museo Civico di Storia Naturale di Genova 50: 79-96. Demir H, Topcu A, Tiirkes T (2006) A new species of the genus Xysticus C.L. Koch from Turkey (Araneae: Thomisidae). 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