Crop wild relatives (CWR) represent valuable sources of traits that can enhance the abiotic and biotic stress tolerance of crops. Due to climate change and expansion of land use, they may suffer from population decline, which increases the risk of losing these resources. The in situ conservation of CWR is receiving growing attention to preserve their intraspecific diversity. In contrast to the safeguarding of seeds ex situ, the preservation of taxa in their natural habitat facilitates the development of new adaptations to changing environmental conditions. However, studies aiming at the establishment of their in situ conservation are rare. Therefore, we identified and genotyped Helosciadium nodiflorum occurrences in Germany. Helosciadium nodiflorum is an endangered CWR of cultivated celery (Apium graveolens). For 20 occurrences the number of individuals, the number of sexually reproducing individuals, and potential threats were recorded. Genetic differences were examined using 17 newly developed simple sequence repeats (SSR). Population genetic analyses, like the inference of the compositional genetic differentiation, of population structure, and a Discriminant Analysis of Principal Components were conducted. The analyses revealed that most occurrences were genetically distinct. Geographically closely located populations are often also genetically more similar. Several populations revealed a composite genome indicative of hybridizations between distinct populations. Populations that experienced fixation were also identified. Based on the genetic diversity, the location in different ecogeographic units, and the support of local stakeholders, we recommend the establishment of 15 genetic reserves for H. nodiflorum in Germany.