Abstract Genital morphology, a cornerstone in taxonomy that predates Linnaeus’s Systema Naturae, is vital for species delimitation. However, the widely accepted paradigm that genitalia are taxonomically informative lacks robust testing between closely related species, and supporting evidence is often limited to taxonomic literature in which genitalia are assumed a priori to be species-specific. The cosmopolitan ant genus Nylanderia Emery includes 123 described species, with most in the Neotropics still undescribed. Workers are often morphologically cryptic, and males are rare in collections but required for morphological delimitation. Using Ultraconserved Elements (UCEs) from 236 samples, including 53 Neotropical Nylanderia species, we reconstructed a phylogenetic framework to compare the genitalia (gonopods) of males collected alongside workers. We used geometric morphometrics on images of slide-mounted genitalia from 16 species and nano-CT scans of Nylanderia fulva (Mayr) and Nylanderia pubens (Forel) genital capsules and interpreted results considering phylogeny under maximum likelihood and the multispecies coalescent. We found strong morphological and molecular support for 2 distantly related American clades, identifiable by gonopod shape, with significant differences observed among most species. Three previously reported COI clades of N. fulva were not supported as monophyletic, nor were their gonopods significantly different. However, N. pubens was supported as distinct by all phylogenetic and 3DGM results. Our findings emphasize the importance of male genitalia for delimiting species boundaries and revising Neotropical Nylanderia. Given their importance, particularly in morphologically cryptic taxa, we recommend a greater focus on linking male and worker phenotypes, which can be facilitated through comprehensive nest series collection. 
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                            Using an integrative taxonomic approach to delimit a sibling species, Mycetomoellerius mikromelanos sp. nov. (Formicidae: Attini: Attina)
                        
                    
    
            The fungus-growing ant Mycetomoellerius (previously Trachymyrmex ) zeteki (Weber 1940) has been the focus of a wide range of studies examining symbiotic partners, garden pathogens, mating frequencies, and genomics. This is in part due to the ease of collecting colonies from creek embankments and its high abundance in the Panama Canal region. The original description was based on samples collected on Barro Colorado Island (BCI), Panama. However, most subsequent studies have sampled populations on the mainland 15 km southeast of BCI. Herein we show that two sibling ant species live in sympatry on the mainland: Mycetomoellerius mikromelanos Cardenas, Schultz, & Adams and M . zeteki . This distinction was originally based on behavioral differences of workers in the field and on queen morphology ( M . mikromelanos workers and queens are smaller and black while those of M. zeteki are larger and red). Authors frequently refer to either species as “ M . cf. zeteki ,” indicating uncertainty about identity. We used an integrative taxonomic approach to resolve this, examining worker behavior, chemical profiles of worker volatiles, molecular markers, and morphology of all castes. For the latter, we used conventional taxonomic indicators from nine measurements, six extrapolated indices, and morphological characters. We document a new observation of a Diapriinae (Hymenoptera: Diapriidae) parasitoid wasp parasitizing M . zeteki . Finally, we discuss the importance of vouchering in dependable, accessible museum collections and provide a table of previously published papers to clarify the usage of the name T . zeteki . We found that most reports of M . zeteki or M . cf. zeteki —including a genome—actually refer to the new species M . mikromelanos . 
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                            - Award ID(s):
- 1927161
- PAR ID:
- 10284835
- Date Published:
- Journal Name:
- PeerJ
- Volume:
- 9
- ISSN:
- 2167-8359
- Page Range / eLocation ID:
- e11622
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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