Large-scale DNA barcoding of ants from Ecuador [results of the first focus of the project] G. Sonet1, T. Delsinne2, D.A. Donoso2, A. Troya3, M. Leponce4, W. Dekoninck5, Z.T. Nagy1, J. Lattke6 T. Backeljau1,7 1 Royal Belgian Institute of Natural Sciences, OD Taxonomy and Phylogeny & JEMU, 29 Vautierstraat, B-1000 Brussels, Belgium; contact: [email protected]; 2 Museo de Colecciones Biológicas MUTPL, Departamento de Ciencias Naturales, Universidad Técnica Particular de Loja, Loja, Ecuador; 3 Escuela Politécnica Nacional; Instituto de Ciencias Biológicas, Sección de Entomología, Ladrón de Guevara E11-253, EC170109, Quito, Ecuador; 4 Royal Belgian Institute of Natural Sciences, OD Natural Environment, Biodiversity Monitoring & Assessment (BIOMON), 29 Vautierstraat, B-1000 Brussels, Belgium; 5 Royal Belgian Institute of Natural Sciences, Scientific Service Heritage, 29 Vautierstraat, B-1000 Brussels, Belgium; 6 Universidad Nacional de Loja, Programa Biodiversidad, Bosques y Servicios Ecosistémicos, Av. Pío Jaramillo Alvarado y Reinaldo Espinosa, La Argelia, Ecuador; 7 University of Antwerp, Evolutionary Ecology Group, 171 Groenenborgerlaan, B-2020 Antwerp, Belgium. Introduction Some ant taxa are morphologically difficult to identify due To facilitate species identification, a joint project was launched First focus of this project3 to caste polymorphism, overlap between intraspecific in 2015, aiming at creating a reference collection of DNA Ecuador has a great habitat diversity and high species We investigated 8 newly collected Leptanilloides variation and interspecific differentiation, uniform worker barcodes for every Ecuadorian ant species or morphospecies. richness but its ant fauna is poorly known. (Formicidae: Dorylinae) specimens. These ants (12 morphology, lack of recent taxonomic revisions, etc. 2 Distribution of ant locality records in Ecuador species) are rarely collected because of their Below: morphologically similar Solenopsis species with subterranean foraging habits. diverging COI sequences (>10%)1. Localities where Leptanilloides specimens were collected before4. Newly collected Leptanilloides spp. © 2002-2015 AntWeb4 Results (morphology) Results (COI barcode – 658 bp) Results (wingless – 412 bp) Three putative new species were recognized in the newly collected Pairwise p-distances among the 3 putative new species: 18.8-21.7%. Pairwise p-distances between the nuclear wingless sequences of L. sp2 and 5 other material based on 22 measurements and 6-7 diagnostic characters. Divergences with the 2 Leptanilloides species in BOLD (6 specimens): 18.0-21.8%. formally described Leptanilloides species from GenBank: 6.4-10.4%. This is in the Below: NJ tree with p-distances & bootstrap values (>80%). interspecific range of Leptanilloides species (2.0 – 10.4%). L. sp1 (1 worker) L. sp2 (6 workers) L. sp3 (1 male) Below: NJ tree with p-distances & bootstrap values (>80%). Note that Leptanilloides frontal frontal belongs with Amyrmex to a monophyletic clade on the basis of the recent molecular phylogeny of Dorylinae5. 4052302 4052312 4052311 L. sp2 (worker) 100 4060602 4052314 KJ523541 & DQ353107 L. nubecula 93 4060603 lateral 4870101 L. sp3 (male) 99 DQ353106 L. nomada lateral ASLAM1144-11 L. gracilis KF371235 L spMAS001 100 AY867429 L. mckennae 100 ASLAM1070-11 L. gracilis 4052311 L. sp2 (worker) ASLAM381-11 L. gracilis KJ523539 L. femoralis JF863464 BOLD:AAQ1360 4006302 L. sp1 (worker) 98 FJ588490 Amyrmex sp. BR01 KJ523540 L. gracilis dorsal DQ353302 L. nubecula dorsal AY233723 Cylindromyrmex striatus 100 KJ523535 Cylindromyrmex meinerti AY233640 Cylindromyrmex striatus 0.02 AY867426 Cylindromyrmex striatus 100 0.01 Discussion • COI and wingless divergences correlate with the morphometric distinction of the 3 putative new morphospecies. Yet, further DNA comparisons are needed with Leptanilloides species that are not yet barcoded. • The taxonomy of Leptanilloides is based on the worker morphology L. sp1 and L. sp2 are good new species candidates. • Male L. sp3 could not yet be assigned L. sp3 is either a new species or the male of a species with unknown male caste. References and acknowledgments 1 Delsinne T, Sonet G, Nagy ZT, Wauters N, Jacquemin J & Leponce M. 2012. Invertebrate Systematics, 26, 457 – 469. 2 Salazar F, Reyes-Bueno F, Sanmartin D & Donoso D.A. 2015. Sociobiology 62(2): 132 – 162. 3 Delsinne T, Sonet G, Donoso D.A. (submitted). European Journal of Taxonomy. 4 AntWeb. Available from http://www.antweb.org. Accessed 5 August 2015. 5 Brady SG, Schultz TR, Fisher BL & Ward PS. 2014. BMC Evolutionary Biology 14, 93. We thank the Ecuadorian Senescyt for financial support (Prometeo grant), Belspo Action 1, FWO, BeBoL and EDIT for their support, the Copalinga Private Reserve, the ECSF-Estación Científica San Francisco, Naturaleza y Cultura Internacional, Isabelle Bachy, Julien Cillis, Yves Laurent for their help with ant digitization, Tania Milena Arias-Penna, Delia Magaly Montalván Carrión, and Jaime Peña for assistance during fieldwork .