Combining the resurrection approach with transplant experiments to investigate adaptation of plant populations to environmental change

Recent climatic changes, such as more frequent droughts and heatwaves, can lead to rapid evolutionary adaptations in plant populations. Such rapid evolution can be investigated using the resurrection approach by comparing plants raised from stored ancestral and contemporary seeds from the same population. This approach has so far only been used in common garden experiments, allowing to reveal genetic differentiation but not adaptation. In this study, we performed a novel approach by testing for evolutionary adaptation in natural plant populations using a resurrection study in combination with in situ transplantations. We cultivated seedlings from ancestors (23-26 years old) and contemporary descendants of three perennial species (Melica ciliata, Leontodon hispidus and Clinopodium vulgare) from calcareous grasslands in the greenhouse and transplanted them back to their collection sites. In addition, we sowed seeds of ancestors and descendants of two species (L. hispidus and C. vulgare) to the collection sites in order to investigate germination rates. In transplanted M. ciliata seedlings, we observed lower mortality, larger plant size and more leaves in descendants compared to ancestors. This indicates that descendants are better adapted than ancestors to the current environmental conditions, which proved to be exceptionally hot and dry during the study period. Descendants of C. vulgare seedlings tended to be smaller and descendants of L. hispidus seedlings produced less leaves compared to their ancestors in their contemporary environmental conditions. In C. vulgare and L. hispidus, we found evolution towards faster germination, and especially descendant seeds of C. vulgare were better adapted to the unfavourable conditions during the experimental period. Concluding, we demonstrate that our novel approach to combine resurrection ecology with transplant experiments is a promising avenue to rigorously test for evolutionary adaptations in changing environments.