Genetic monitoring of Antennaria dioica
Antennaria dioica (L.) Gaertn. is a diploid (2n = 28; Bayer, 1984), dioecious, perennial hemicryptophyte inhabiting dry, nutrient-poor meadows, grasslands, and open woodlands. It is protected by law, nationally endangered and threatened with extinction in the study region of central Germany, mainly due to land-use change, eutrophication, and shrub encroachment.  We collected plant material in populations in Saxony-Anhalt, Germany, surveying all sites reported in the past 20 years and sites previously sampled by Rosche et al. (2014) in Bavaria, Brandenburg, Saxony, Thuringia, and the Czech region Ústecký kraj. To trace spatial-temporal changes, herbarium specimens from nine herbaria were examined and in total, 71 herbarium leaf samples were used for DNA extraction. DNA from 2022/2023 samples was extracted using the NucleoSpin Plant II kit (Macherey-Nagel); 2010 samples had been extracted via the ATMAB method (Stein et al., 2014) and stored at –80 °C. Herbarium DNA was extracted using the CTAB method (Doyle & Doyle, 1990) with a sorbitol prewash (Inglis et al., 2018). The double digest Restriction-site Associated DNA (ddRAD) protocol from Peterson et al. (2012) was applied as described in Durka et al. (2025) to identify SNP (single nucleotide polymorphism) markers. In total, 480 samples were processed: 43 historical, 139 from 2010, and 298 from 2022/2023. In short, 100 ng of DNA were restricted with EcoRI and MspI enzymes, adapters containing one of 96 7-bp barcodes were ligated to fragments, which were then pooled, size selected to 350-450 bp, PCR-amplified, thereby introducing a plate-specific 5 bp index, with intermediate and final cleanup steps. Multiple plates were finally equimolarly multiplexed for sequencing on an Illumina Novaseq 6000, resulting in an average of 4,483,114 sequences per sample. Demultiplexing of the raw reads was performed using process_radtags from the Stacks pipeline (Catchen et al. 2013, doi:10.1534/g3.111.000240).
Antennaria dioica (L.) Gaertn. is a diploid (2n = 28; Bayer, 1984), dioecious, perennial hemicryptophyte inhabiting dry, nutrient-poor meadows, grasslands, and open woodlands. It is protected by law, nationally endangered and threatened with extinction in the study region of central Germany, mainly due to land-use change, eutrophication, and shrub encroachment.  We collected plant material in populations in Saxony-Anhalt, Germany, surveying all sites reported in the past 20 years and sites previously sampled by Rosche et al. (2014) in Bavaria, Brandenburg, Saxony, Thuringia, and the Czech region Ústecký kraj. To trace spatial-temporal changes, herbarium specimens from nine herbaria were examined and in total, 71 herbarium leaf samples were used for DNA extraction. DNA from 2022/2023 samples was extracted using the NucleoSpin Plant II kit (Macherey-Nagel); 2010 samples had been extracted via the ATMAB method (Stein et al., 2014) and stored at –80 °C. Herbarium DNA was extracted using the CTAB method (Doyle & Doyle, 1990) with a sorbitol prewash (Inglis et al., 2018). The double digest Restriction-site Associated DNA (ddRAD) protocol from Peterson et al. (2012) was applied as described in Durka et al. (2025) to identify SNP (single nucleotide polymorphism) markers. In total, 480 samples were processed: 43 historical, 139 from 2010, and 298 from 2022/2023. In short, 100 ng of DNA were restricted with EcoRI and MspI enzymes, adapters containing one of 96 7-bp barcodes were ligated to fragments, which were then pooled, size selected to 350-450 bp, PCR-amplified, thereby introducing a plate-specific 5 bp index, with intermediate and final cleanup steps. Multiple plates were finally equimolarly multiplexed for sequencing on an Illumina Novaseq 6000, resulting in an average of 4,483,114 sequences per sample. Demultiplexing of the raw reads was performed using process_radtags from the Stacks pipeline (Catchen et al. 2013, doi:10.1534/g3.111.000240).
HELMHOLTZ CENTRE FOR ENVIRONMENTAL RESEARCH - UFZ.