biostudies-literatureGutierrez-Rodriguez JDoctores Junta de AndalucíaMinisterio de Ciencia e Innovación, SpainJuan de la Cierva-Formación, Ministerio de Ciencia e Innovación, SpainConsejo Nacional de Humanidades, Ciencias y TecnologíasDoctores Junta de Andalucía (FEDER EU/Consejería de Economía, Conocimiento, Empresas y Universidad, Junta de Andalucía)33883https://www.ebi.ac.uk/biostudies/studies/S-EPMC12484681biostudies-literatureUnknown15(1)Curated DNA barcode reference libraries are crucial for advancing environmental DNA (eDNA) studies, monitoring biological invasions, reliable biodiversity assessments, accurate species identification, etc. However, DNA barcode databases remain highly incomplete for most invertebrate taxa. In this study, we present the most comprehensive reference library to date for the family Tettigoniidae (Orthoptera) from the Iberian Peninsula-the most species-rich orthopteran family globally, with over 8,000 valid species. We generated 402 new DNA barcodes from at least 121 tettigoniid species from the Iberian Peninsula and integrated these with 169 previously published sequences. The resulting dataset comprises 571 barcoded specimens, representing 49 genera and 123 species, including many recently desScientific reportsDNA barcode reference library of bush-crickets (Orthoptera, Tettigoniidae) from the Iberian Peninsula.PMC12484681PID2019-110243GB-100/AEI/10.13039/501100011033DOC_0066858548FJC2018-035899-IZaldivar-Riveron AGutierrez-Rodriguez JBarranco PGarcia-Paris MLopez-Estrada EKfalseDNA barcode reference library of bush-crickets (Orthoptera, Tettigoniidae) from the Iberian Peninsula.Curated DNA barcode reference libraries are crucial for advancing environmental DNA (eDNA) studies, monitoring biological invasions, reliable biodiversity assessments, accurate species identification, etc. However, DNA barcode databases remain highly incomplete for most invertebrate taxa. In this study, we present the most comprehensive reference library to date for the family Tettigoniidae (Orthoptera) from the Iberian Peninsula-the most species-rich orthopteran family globally, with over 8,000 valid species. We generated 402 new DNA barcodes from at least 121 tettigoniid species from the Iberian Peninsula and integrated these with 169 previously published sequences. The resulting dataset comprises 571 barcoded specimens, representing 49 genera and 123 species, including many recently des2025-01-01T00:00:00Z2025 Sep2026-06-04T01:44:11.625Z2026-05-04T03:13:23.006ZS-EPMC124846814102801710.1038/s41598-025-06695-2