Phylogenomic placement and revision of Iranattus Prószyński, 1992 jumping spiders (Salticidae, Plexippini, Plexippina)

The jumping spider genus Iranattus Prószyński, 1992, distributed from Africa to southwestern Asia, has been placed within the Har - mochirina because of their male palp structures and elongated third legs. Here, we present phylogenomic evidence that it belongs instead to the subtribe Plexippina, further supported by the presence of two coupling pockets in the female epigyne. In this study, we redescribe I. principalis (Wesołowska, 2000) and I. rectangularis Prószyński, 1992. Additionally, the female of I. rectangularis , the type species of the genus, is described for the first time, and we report its range extension east to India.


Introduction
When Prószyński (1992) originally described the jumping spider genus Iranattus Prószyński, 1992, based on a single male specimen from Iran, he characterized it by features such as a simple tegulum (bulbus) and embolus, unusual cymbial apophysis, and an extraordinarily long pair of legs (which his text erroneously states are the fourth pair, but which in fact are the third, as in his figures [35][36].These traits led Maddison (2015) to place it within the Harmochirina, some of which have very long third legs (e.g., Neaetha Simon, 1885), and some of which (e.g., Pellenes limbatus Kulczyński, 1895) have an apophysis on the male cymbium very similar to that of Iranattus.A relationship with Harmochirines was suggested by Wesołowska (2000), who, when describing Monomotapa Wesołowska, 2000 (later synonymized with Iranattus; Prószyński 2017), commented on its similarity in body and leg lengths with the harmochirines Neaetha Simon, 1885, andPellolessertia Strand, 1929.Subsequent studies and new material now give the opportunity to reconsider the phylogenetic placement of Iranattus, currently composed of two species (World Spider Catalog 2024).Females were unknown until Wesołowska and Russell-Smith's (2022) recent redescription of the African I. principalis (Wesołowska, 2000), known from Côte d'Ivoire, Nigeria, and Zimbabwe (Wesołowska 2000; Wesołowska andRussell-Smith 2011, 2022).We have recently collected I. rectangularis Prószyński, 1992, in India, allowing us to not only characterize it through living photographs and natural history information but also to describe its female for the first time and to gather genetic data.We set out to clarify its placement phylogenomically using ultraconserved element (UCE) data and with information on female genitalic morphology.Additionally, we provide a comprehensive generic diagnosis and redescribe both species.

Materials examined
The specimens of I. rectangularis were recently collected from the Desert National Park, Rajasthan, India.They are currently housed in the collection of the Centre for Animal Taxonomy and Ecology (CATE), Christ College, Kerala, with plans for eventual transfer to the Research Collections at the National Centre for Biological Sciences (NCBS), Bengaluru, Karnataka, India (http:// collections.ncbs.res.in), for permanent deposition.NRC-AA-#### represent NCBS voucher codes of I. rectangularis used for taxonomic work, where #### represents a four-digit number.
The I. principalis specimens used in this study were in vials in a large jar of poorly labeled salticid specimens in the Natural History Museum, London (NHMUK).All the vials in the jar contained, typically, African salticids.Their labels bore only codes of the form "PNB ###", where ### is a two-or three-digit number.We interpret these to likely be Lamotte's collection from Parc Nacional Banco (hence, "PNB"), Côte d'Ivoire, from which Wanless (1985) cites similar code labels under Sonoita lightfooti Peckham & Peckham, 1903, e.g., PNB 179, PNB 146.Some specimens are identified by voucher codes of the form DDKM21.###, where ### is a three-digit number.

Morphology
We examined and photographed ethanol-preserved specimens using an Olympus OM-D E-M10 II camera mounted on an Olympus SZX12 or a Leica DMC4500 camera attached to a Leica M205 C stereoscope.We used a drawing tube attached to a Nikon ME600L compound microscope to prepare illustrations of I. principalis.We used clove oil for clear viewing of epigyne after digesting the internal epigynal soft tissues with pancreatin.We stacked photographs using Helicon Focus 7.6.6Pro.We prepared the drawings of I. rectangularis specimens by digitally tracing the photographs.
Descriptions of color patterns are based on ethanol-preserved specimens.Carapace length is measured from the base of the anterior median eyes to the posterior margin of the carapace medially, while abdomen length is measured from the anterior to the end of the anal tubercle.All measurements are in millimeters.Leg measurements are represented as follows: total length (femur, patella, tibia, metatarsus, and tarsus).Abbreviations used here are as follows: CO, copulatory opening; ECP, epigynal coupling pocket; PME, posterior median eye; PLE, posterior lateral eye; RTA, retrolateral tibial apophysis.

Taxon sampling for phylogenomic analysis
To test the phylogenetic placement of Iranattus, molecular data was gathered for I. rectangularis and added to Marathe et al.'s (2024) UCE phylogenomic dataset, which included 15 plexippines, two harmochirines, and one salticine.Because Iranattus's former placement in the Harmochirina was based in part on some Pellenes having a similar cymbial apophysis, one such Pellenes (Pellenes limbatus) was added to the dataset to give the harmochirines the best chance to capture Iranattus in the phylogenetic analysis.An extra outgroup taxon, Chrysilla volupe (Karsch, 1879), was also added.The total set of 21 species used in the phylogenomic analysis, with their taxonomic authority indicated, is listed in Table 1.

Ultraconserved element (UCE) data
Molecular data was gathered for UCE loci using target enrichment sequencing methods (Faircloth 2017), using the RTA_v2 probeset (Zhang et al. 2023), and following the protocols of Marathe et al. (2024).

Phylogenetic analysis
Maximum-likelihood phylogenetic and bootstrap analyses were performed with IQ-TREE v. 2.2.0 (Nguyen et al. 2015) using the Zephyr v. 3.31 package (Maddison and Maddison 2023a) in Mesquite v. 3.81 (Maddison and Maddison 2023b) on the concatenated, unpartitioned UCE dataset with 20 taxa.For the phylogenetic tree inference, the option -m TEST (standard model selection followed by tree inference, edge-linked partition model, no partition-specific rates) was used with 10 search replicates.For the bootstrap analysis, a single IQ-TREE search was used for each of the 1000 search replicates.

Data availability
The raw sequence reads obtained from UCE capture are stored within the Sequence Read Archive (BioProject: https://www.ncbi.nlm.nih.gov/bioproject/1101580), and their accession numbers are listed in Table 1.The UCE loci matrices from SPAdes assemblies, pre-Gblocks, and the concatenated matrices used for phylogenetic and bootstrap analysis, along with trees, are available on the Dryad data repository (https://doi.org/10.5061/dryad.ht76hdrpz).

Phylogenetic results
Table 2 lists the sequence data recovered from the 21 taxa.3398 UCE loci were initially recovered.Of these, 3140 remained after removing those represented in fewer than 10 taxa, and 3104 remained after removing those suspected to include paralogies on branch lengths.These were concatenated into the final matrix, whose aligned length is 2779616 base pairs, in which each taxon had on average ~2.2 million base pairs of sequence data (min. 985191, max. 2462121).
The phylogenetic results are shown in Fig. 1.The reciprocal monophyly of the subtribes Plexippina and Harmochirina  is consistent with previous molecular phylogenetic studies with both Sanger sequencing and UCEs (Maddison and Hedin 2003;Maddison et al. 2008;Bodner and Maddison 2012;Marathe et al. 2024).The phylogenetic structure within Plexippina is largely consistent with Marathe et al. (2024) and has generally high bootstrap values.
Iranattus is nestled well within Plexippina, placed as a sister lineage to Evarcha Simon, 1902 sensu lato (see Fig. 1).The harmochirine included in the analysis with a similar cymbial apophysis, Pellenes limbatus, is placed as expected within the harmochirines.Thus, the similarities between Iranattus and harmochirines noted by Wesołowska (2000) and Maddison (2015) are convergences.
The placement of Iranattus in the Plexippina is also supported by the form of the epigyne.Wesołowska and Russell-Smith (2022) report a pair of coupling pockets in I. principalis, one on either side of a central atrium housing the copulatory openings, the same as we have found in I. rectangularis (Figs 22,28).This arrangement is discordant with that of harmochirines, which have a single epigynal coupling pocket placed centrally, anterior to the margin, flanked by copulatory openings on either side.Two pockets are typical, however, for members of the Plexippina (e.g., Evarcha, Baryphas Simon, 1902;Pancorius Simon, 1902;Telamonia Thorell, 1887;Vicirionessa Wesołowska & Russell-Smith, 2022).
We therefore recognize Iranattus as a member of the subtribe Plexippina.

Iranattus Prószyński, 1992
Figs 2-41 Iranattus Prószyński, 1992: 97-98, f. 35-40. Monomotapa Wesołowska, 2000: 159, f. 42-46 (synonymized by Prószyński, 2017: 36.).Prószyński, 1992.Species included.Iranattus principalis (Wesołowska, 2000); Iranattus rectangularis Prószyński, 1992.Table 2. Specifics of molecular data used for this phylogenomic analysis.Molecular data was generated based on the RTA_v2 probeset."SRA" is the Sequence Read Archive accession number available through NCBI; "Reads pass QC" is the number of reads after the removal of adapter contamination and low-quality bases using Illumiprocessor; "Total UCE loci" is the total number of UCE loci recovered with RTA_v2 probeset; "After paralogy filter" is the number of UCE loci after deletion of suspected paralogous loci based on branch length ratios; "In at least 10 taxa" is the number of UCE loci in at least 10 or more taxa after branch length criteria; "Filtered UCE sequence length" is the concatenated sequence length of filtered UCE loci; "Total loci" is the number of UCE loci represented among all taxa.Diagnosis.The remarkably long third legs of Iranattus (Figs 15,18,30,32) and scoop-shaped cymbial apophysis (Fig. 4) differentiate it from all other plexippines.The very robust carapace, bulging outward at the PLE and bearing the PLEs on tubercles, is unusual but shared also with Afrobeata Caporiacco, 1941, andVailimia Kammerer, 2006.Vailimia especially might be confused with Iranattus, as they share erect hairs on the carapace (see Figs 34,38,41) and a compact crouch stance, but, besides the cymbial apophysis and long third legs, Iranattus also has a shorter embolus lacking membrane (membrane-accompanied long embolus in Vailimia), a short RTA (long and curved in Vailimia), and two distinct deep conical ECPs (absent in Vailimia).From Afrobeata, Iranattus differs in having longer third legs, a cymbial apophysis (lacking in Afrobeata), a shorter embolus (longer in Afrobeata), a simple short RTA (bifurcated in Afrobeata), shorter copulatory ducts (long in Afrobeata), and deep conical ECPs (shallow in Afrobeata).Some other plexippines have cymbial apophyses (Plexippoides Prószyński, 1984;Epeus Peckham & Peckham, 1886;and Erasinus Simon, 1899), but their apophyses are different in shape-in Iranattus, a long, broad blade with a rounded tip, concave in front so as to form a scoop; in Plexippoides, sharply pointed, for example.

Materials examined.
In NHMUK, lacking complete labels.These are likely from Parc Nacional Banco, Côte d'Ivoire (see "Materials examined" for explanation).

2♀♀ (PNB21
Diagnosis.Larger than I. rectangularis, with an almost ovoid tegulum with a less prominent shoulder, RTA slightly bent near the tip (Figs 10,11), and a multi-chambered spermatheca sandwiched between copulatory ducts dorsally and the epigynal plate ventrally.
Natural history.Iranattus rectangularis was collected from the branches of non-native Vachellia tortilis alongside artificial water canals in the Desert National Park, a xeric and desert ecosystem located in Rajasthan, India (Figs 42,43).The mosaic of orange, black, and grey body coloration helps them blend in with the branches, making them inconspicuous, except that in the field, the orangish faces of males (Fig. 34) sometimes stood out.
Discussion.Iranattus rectangularis is reported for the first time east of Iran, in western India.This seemingly 'disjunct' distributional pattern is quite possibly due to a lack of collecting between the sites and mirrors that of Stenaelurillus marusiki Logunov, 2001 (Salticidae: Aelurillina), where the type locality of S. marusiki is Iran.However, it has been reported much farther southeast in Maharashtra, India (Marathe et al. 2022).With the transfer of Iranattus to Plexippina, the subtribe now contains 35 genera, and the number of plexippines in India stands at 47 species and 18 genera.

Table 1 .
Specimens used in phylogenomic analysis.
Figure1.The IQ-TREE-based maximum-likelihood tree, represented here, is the best of 10 replicates, inferred from a concatenated dataset of 3104 UCE loci.The numbers at the nodes are the percentage recovery of the clade based on 500 bootstrap replicates.Iranattus rectangularis is recovered distantly from the subtribe Harmochirina and placed as the sister lineage to Evarcha sensu lato within the subtribe Plexippina.