Aims Functional traits are supposed to play an important role in determining the colonization success of new species into established communities. Short-term experimental studies have documented higher resistance of more diverse grasslands against colonization by new species. However, little is known about which traits colonizers should have to successfully invade diverse plant communities in the longer term and how community history may modify the resistance of diverse communities against colonization. Methods In a grassland biodiversity experiment (Jena Experiment) established with different species richness (SR; 1, 2, 4, 8 and 16) and functional group (FG) number and composition (1 to 4; legumes, grasses, small herbs, tall herbs), we studied colonization of naturally dispersed species in split-plots (i) with different duration of weeding (never weeded, weeded for 3 or 6 years and then un-weeded for 1 year) and (ii) with different duration of colonization (7 years, 4 years and 1 year after cessation of weeding). Important Findings Resistance against colonization by new species declined with increased duration of weeding (on average 13, 17 and 22 colonizer species in 1-, 4- and 7-year-old communities, respectively). Communities established at low diversity accumulated more colonizer species with a longer duration of weeding than more diverse communities. Duration of colonization had only small effects on the number of colonizer species. Colonizers with early successional traits, i.e. annual life cycle, reproduction by seeds, small seeds, long-lived seeds and an earlier start of a longer flowering period, were favoured in species-poor newly established experimental plant communities (short duration of weeding) and early after cessation of weeding (short duration of colonization). A change from early- to mid-successional traits, i.e. taller growth, perennial life cycle, vegetative reproduction, characterized colonization at increased plant diversity and in communities with legumes or without grasses. Legume absence/grass presence and increased duration of weeding led to a shift in colonizer strategies from rapid nutrient uptake and cycling (higher specific leaf area) to nutrient retention and symbiotic N 2 fixation. Our study shows that non-random trait spectra of naturally dispersed colonizers encompass trade-offs between different functions (reproduction, persistence, growth) reflected in a change from early- to mid-successional traits at increasing plant diversity, with a longer duration of weeding and a longer time of colonization.