Resumen: Climate change and invasive species are leading drivers of biodiversity loss, with island ecosystems being especially vulnerable. South Georgia, a remote sub-Antarctic island, is 170 km long with approximately 30,000 ha of vegetated coastal areas, as snow and ice dominate the inland regions. Human activities on the island have historically introduced non-native species, resulting in 41 introduced vascular plant species compared with only 24 native ones. To address this imbalance, the South Georgia Non-Native Plant Management Strategy was implemented (2016–2020) to control non-native plant populations. We assessed emergent seedlings from South Georgia soil samples and wind-dispersed seeds to determine which species persist in the soil seed bank and contribute to dispersal. Using a molecular barcoding approach, we evaluated traditional markers (rbcL and matK) and optimized a high-throughput Angiosperms353 sequencing pipeline for accurate seedling identification. We generated a reference library covering all native and non-native species and applied this to 1,498 emergent seedlings and 737 trapped seeds. Molecular barcoding identified 21 species, including 10 non-natives and 11 natives. Strikingly, 84% of emergent seedlings were non-native, with Class III invasive species (Cerastium fontanum, Poa annua, Taraxacum officinale) dominating across most sites and in all wind traps. By contrast, Class I and II species occurred rarely and only at a few sites, indicating that management efforts have substantially reduced their spread, though viable seeds persist in the soil. These findings highlight both the continued threat from persistent seed banks of dominant invaders and the value of molecular barcoding for long-term monitoring. Our approach provides a framework for biosecurity and restoration management in South Georgia and other vulnerable ecosystems under climate change pressures. Idioma: Inglés DOI: 10.1111/1755-0998.70068 Año: 2025 Publicado en: Molecular Ecology Resources (2025), e70068 [25 pp.] ISSN: 1755-098X Financiación: info:eu-repo/grantAgreement/ES/AEI/RYC2023-042611-I Tipo y forma: Artículo (Versión definitiva)