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Supplementary material 15 from: Hita Garcia F, Fischer G, Liu C, Audisio TL, Economo EP (2017) Next-generation morphological character discovery and evaluation: an X-ray micro-CT enhanced revision of the ant genus Zasphinctus Wheeler (Hymenoptera, Formicidae, Dorylinae) in the Afrotropics. ZooKeys 693: 33-93. https://doi.org/10.3897/zookeys.693.13012

Video 2 : Explanation note: Zasphinctus obamai sp. n. paratype worker (CASENT0764127). 3D rotation video of volumetric surface rendering of mesosoma (head and metasoma mostly virtually removed).

Supplementary material 11 from: Hita Garcia F, Fischer G, Liu C, Audisio TL, Economo EP (2017) Next-generation morphological character discovery and evaluation: an X-ray micro-CT enhanced revision of the ant genus Zasphinctus Wheeler (Hymenoptera, Formicidae, Dorylinae) in the Afrotropics. ZooKeys 693: 33-93. https://doi.org/10.3897/zookeys.693.13012

3D PDF 11 : Explanation note: Zasphinctus wilsoni sp. n. holotype worker (MCZ-ENT-00512764). 3D PDF of volumetric surface rendering model of mesosoma (head and metasoma mostly virtually removed).

Supplementary material 12 from: Hita Garcia F, Fischer G, Liu C, Audisio TL, Economo EP (2017) Next-generation morphological character discovery and evaluation: an X-ray micro-CT enhanced revision of the ant genus Zasphinctus Wheeler (Hymenoptera, Formicidae, Dorylinae) in the Afrotropics. ZooKeys 693: 33-93. https://doi.org/10.3897/zookeys.693.13012

3D PDF 12 : Explanation note: Zasphinctus wilsoni sp. n. holotype worker (MCZ-ENT-00512764). 3D PDF of volumetric surface rendering model of metasoma (virtually separated from mesosoma and most of legs virtually removed).

Supplementary material 13 from: Hita Garcia F, Fischer G, Liu C, Audisio TL, Economo EP (2017) Next-generation morphological character discovery and evaluation: an X-ray micro-CT enhanced revision of the ant genus Zasphinctus Wheeler (Hymenoptera, Formicidae, Dorylinae) in the Afrotropics. ZooKeys 693: 33-93. https://doi.org/10.3897/zookeys.693.13012

3D PDF 13 : Explanation note: Zasphinctus sarowiwai sp. n. worker (CASENT0764652). 3D PDF of volumetric surface rendering of segmented mouthparts excluding mandibles (green= maxillae; yellow=labrum; orang=labium).

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Data from: Next-generation morphological character discovery and evaluation: an X-ray micro-CT enhanced revision of the ant genus Zasphinctus Wheeler (Hymenoptera, Formicidae, Dorylinae) in the Afrotropics

New technologies for imaging and analysis of morphological characters offer opportunities to enhance revisionary taxonomy and better integrate it with the rest of biology. In this study, we revise the Afrotropical fauna of the ant genus Zasphinctus Wheeler, and use high-resolution X-ray microtomography (micro-CT) to analyse a number of morphological characters of taxonomic and biological interest. We recognise and describe three new species: Z. obamai sp. n., Z. sarowiwai sp. n., and Z. wilsoni sp. n. The species delimitations are based on the morphological examination of all physical specimens in combination with 3D scans and volume reconstructions. Based on this approach, we present a new taxonomic discrimination system for the regional fauna that consists of a combination of easily observable morphological characters visible at magnifications of around 80–100 ×, less observable characters that require higher magnifications, as well as characters made visible through virtual dissections that would otherwise require destructive treatment. Zasphinctus are rarely collected ants and the material available to us is comparatively scarce. Consequently, we explore the use of micro-CT as a non-invasive tool for the virtual examination, manipulation, and dissection of such rare material. Furthermore, we delineate the treated species by providing a diagnostic character matrix illustrated by numerous images and supplement that with additional evidence in the form of stacked montage images, 3D PDFs and 3D rotation videos of scans of major body parts and full body (in total we provide 16 stacked montage photographs, 116 images of 3D reconstructions, 15 3D rotation videos, and 13 3D PDFs). In addition to the comparative morphology analyses used for species delimitations, we also apply micro-CT data to examine certain traits, such as mouthparts, cuticle thickness, and thoracic and abdominal muscles in order to assess their taxonomic usefulness or gain insights into the natural history of the genus. The complete datasets comprising the raw micro-CT data, 3D PDFs, 3D rotation videos, still images of 3D models, and coloured montage photos have been made available online as cybertypes (Dryad, http://dx.doi.org/10.5061/dryad.4s3v1).

Fig. 11 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 11. Distribution map, face view, lateral view of petiole-AIV, and dorsal view of AIII-AIV of Syscia madrensis (holotype worker), S. JTL066 (worker, CASENT0610648), S. benevidesae (holotype worker), S. chiapaneca (holotype worker), S. parietalis (holotype worker), S. setosa (holotype worker), S. JTL073 (worker, MCZ-ENT00511569), S. JTL075 (worker, CASENT0601445), S. disjuncta (holotype worker), and S. augustae (worker, CASENT0644275). Species are in order of mean HW, which is shown in the lower left of the distribution map. Scale bars 0.2 mm. On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.

Fig. 10 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 10. Distribution map, face view, lateral view of petiole-AIV, and dorsal view of AIII-AIV of Syscia jennierussae (holotype worker), S. persimilis (holotype worker), S. JTL018 (worker, JTLC000013995), S. machaquila (holotype worker), S. murillocruzae (holotype worker), S. truncata (holotype worker), S. JTL084 (worker, FMNHINS0000095760), S. JTL050 (worker, CASENT0249320), S. JTL082 (worker, CASENT0617700), and S. honduriana (lectotype worker). Species are in order of mean HW, which is shown in the lower left of the distribution map. Scale bars 0.2 mm. On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.

Fig. 7 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 7. Phylogenetic relationships among COI barcode sequences for Syscia. Red samples were sequenced for UCEs. Black samples were downloaded from the BOLD database.The tree was inferred using IQ-TREE with the data partitioned by codon position. Black circles on nodes indicate high support, which we define as ≥95% ultrafast bootstrap support and ≥95% SH-like branch support. Clades of named species are shaded as a visual aid, with gray outlines indicating non-monophyly of species.

Fig. 1 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 1. Variation in Syscia occipital carina. (A) Flange weakly developed, less visible in face view. (B) Flange strongly developed, easily visible in face view.

Fig. 6 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 6. Phylogeny of New World Syscia, inferred using the program IQ-TREE and 1,388 UCE loci.Two outgroup taxa (two species of Ooceraea) are not shown. Node support values (ultrafast bootstrap/SH-like) <100/100 are depicted with red dots.The imaged specimen is S. ticomontana (CASENT0644376).

Fig. 3 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 3. Variation in Syscia profiles and pilosity. (A) AIII in dorsal view trapezoidal, with convex sides. (B) AIII in dorsal view weakly trapezoidal, with convex sides. (C) AIII in dorsal view trapezoidal, with flat sides. (D) AIV in dorsal view, with convex sides, anterior margin not truncate. (E) AIV in dorsal view, with convex sides, anterior margin moderately truncate. (F) AIV in dorsal view, with nearly flat sides, anterior margin strongly truncate. (G) AIII dorsal profile strongly convex. (H) AIII dorsal profile weakly convex. (I) AIII dorsal profile flat. (J) AIV dorsal profile convex. (K) AIV dorsal profile weakly convex. (L) AIV dorsal profile flat. (A, B, G, J) Standing pilosity long, coarse. (C, H, K) Standing pilosity of medium length and thickness. (I, L) Standing pilosity short, fine.

Fig. 13 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 13. Distribution map, face view, lateral view of petiole-AIV, and dorsal view of AIII-AIV of Syscia latepunctata (holotype worker), S. borowieci (holotype worker), S. volucris (holotype worker), S. JTL076 (queen, CASENT0614221),S. JTL064 (worker, CASENT0631661), S. JTL033 (worker, CASENT0611831),S. grandis (holotype worker), and S. JTL003 (worker, INB0003213589). Species are in order of mean HW, which is shown in the lower left of the distribution map. Scale bars 0.2 mm. On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.

Fig. 9 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 9. Distribution map, face view, lateral view of petiole-AIV, and dorsal view of AIII-AIV of Syscia pervagata (holotype worker), S. peten (holotype worker), S. JTL074 (worker, MCZ-ENT00511564), S. brachyptera (holotype worker), S. valenzuelai (holotype worker), S. JTL071 (worker, FMNHINS0000095759), S. quisquillis (holotype worker), S. sumnichti (holotype worker), S. JTL060 (worker, CASENT0644220), and S. JTL085 (worker, CASENT0602939). Scale bars 0.2 mm. Species are in order of mean HW, which is shown in the lower left of the distribution map. On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.

Fig. 2 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 2. Variation in Syscia subpetiolar process. (A) Subtriangular with flat to concave posterior margin. (B) Subtriangular with convex posterior margin. (C) Subtriangular with small tooth on posterior margin. (D) Subquadrate. (E) Subtriangular with large acute tooth on posterior margin. F. With fenestra and notch on posterior margin.

Fig. 4 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 4. Illustrations of Syscia measurements. HL: head length, HW: head width, MSL: mesosoma length, AIIIL: abdominal tergite III length, AIIIW: abdominal tergite III width, AIVL: abdominal tergite IV length, AIIVW: abdominal tergite IV width.

Fig. 12 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 12. Distribution map, face view, lateral view of petiole-AIV, and dorsal view of AIII-AIV of Syscia tolteca (lectotype worker), S. atitlana (holotype worker), S. lacandona (holotype worker), S. JTL049 (worker, CASENT0644222), S. JTL065 (worker, CASENT0602939), S. amblyogyna (holotype worker), S. ticomontana (holotype worker), S. JTL017 (worker, INB0003693097), S. JTL079 (worker, CASENT0642985), and S. transisthmica (holotype worker). Species are in order of mean HW, which is shown in the lower left of the distribution map. Scale bars 0.2 mm. On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality (not shown for S. tolteca, with type locality 'Guatemala').

Fig. 8 in Integrating UCE Phylogenomics With Traditional Taxonomy Reveals a Trove of New World Syscia Species (Formicidae: Dorylinae)

Fig. 8. Distribution map, face view, lateral view of petiole-AIV, and dorsal view of AIII-AIV of Syscia minuta (holotype worker), S. parva (holotype worker), S. JTL067 (worker, CASENT0644012), S. pollula (holotype worker), S. JTL069 (worker, CASENT0644008), S. JTL068 (queen, CASENT0613276), S. austrella (holotype worker), S. JTL037 (worker, CASENT0635747), S. quisquillis Arizona form (worker, FMNHINS0000095772), and S. boudinoti (holotype worker). Species are in order of mean HW, which is shown in the lower left of the distribution map. Scale bars 0.1 mm for S. minuta to S. JTL037, 0.2 mm for S. quisquillis Arizona form and S. boudinoti. On distribution maps, red dots are sites with UCE sequence data. Red boxes are type locality.

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Fossil Dorylinae CT data

Among social insects, army ants are exceptional in their voracious coordinated predation and nomadic life history: the synthesis of these remarkable ecological traits is referred to as the army ant syndrome. Despite molecular evidence that the army ant syndrome evolved twice during the mid-Cenozoic, once in the Neotropics and once in the Afrotropics, fossil army ants are markedly scarce, comprising a single known species from the Caribbean 16 Ma. Here we report the oldest army ant fossil and the first from the Eastern Hemisphere (EH), Dissimulodorylus perseus, preserved in Baltic amber dated to the Eocene. Using a combined morphological and molecular Ultra Conserved Elements dataset spanning doryline lineages, we find that D. perseus is nested among extant EH army ants with affinities to Dorylus "driver ants." Army ants are characterized by limited extant diversification throughout most of the Cenozoic; the discovery of D. perseus suggests an unexpected diversity of now-extinct army ant lineages in the Cenozoic, some of which were present in Continental Europe.