Dark diversity reveals assembly constraints across trophic levels in large-scale grassland restoration

Abstract

1. Grassland biodiversity restoration efforts have largely focused on re-establishing diverse plant communities, with less attention paid to higher trophic levels such as invertebrates. 2. We established a controlled, field-scale restoration experiment across twelve lowland grassland sites in Switzerland in close collaboration with farmers. We manipulated seed addition (hay transfer, directly harvested seeds, or multiplied seed mixtures) and soil disturbance (shallow or deep tillage), with treatments applied to meadows covering, on average, 0.5 ha. Four to six years after restoration, we quantified the components of species richness change (lost, persisting, gained from site surroundings, and gained from donors), and estimated dark diversity (species that are absent from a location but could potentially occur there) for plants, Auchenorrhyncha and Orthoptera. 3. Compared to plants, invertebrates exhibited greater species turnover, with higher proportions of species lost and gained from the surroundings, but much lower contributions from the donor meadows. However, species persistence was high across taxa. Restoration operations strongly influenced components of species richness change in plants but had weaker effects on invertebrates. In particular, multiplied seed mixtures + deep tillage had the highest proportion of plant species not transferred, suggesting that many species in the seed mixture were poorly suited to local conditions. In both plants and Auchenorrhyncha, components of species richness change were strongly mediated by site characteristics, with initial species richness in both receiver and donor emerging as the strongest predictors. 4. Within each taxonomic group, 41% of species not transferred from the donor belonged to the dark diversity, suggesting substantial unrealized restoration potential. Restoration operations impacted dark diversity for plants and Auchenorrhyncha but not for Orthoptera. In plants, dark diversity was substantially lower under multiplied seed mixtures + deep tillage than under other treatments. The probability of a species belonging to dark diversity was associated with specific leaf area in plants and initial relative abundance in Auchenorrhyncha. 5. Synthesis and applications. Components of species richness change were influenced by both restoration operations and site characteristics, with their relative importance varying among taxa, underscoring the need for multi-taxon restoration strategies.