Flow Cytometry Protocols, Relative Genome Size and Ploidy Levels for 1,103 Species of Non-Apomictic Angiosperms from the Eastern Alps

Flow cytometry provides a reliable and fast method for estimating genome size and ploidy levels in plants. Until recently, most studies employed fresh tissues that usually provide high-quality results. However, the need for fresh tissues limits the use of the method with samples from remote areas or when an extremely high number of samples needs to be processed in a short time. Although there is growing evidence that fixed silica-dried material can be used for ploidy estimation in some taxa, no flora-wide study has been available so far. Here, we provide methodological aspects of an unprecedented study exploring ploidy variation of non-apomictic angiosperms in the Eastern Alps. We have analysed ~45,000 silica-dried samples of 1,134 species using flow cytometry with DAPI as stain. Surprisingly, we were able to obtain ploidy level information from as much as 1,103 (97%) of species. The unsuccessful species included succulent plants of the family Crassulaceae (genera Jovibarba, Rhodiola, Sedum, Sempervivum), the achlorophyllous parasitic or mycoheterotrophic genera Orobanche and Hypopitys, and a handful of others. About 80% of samples were successfully analysed using a single ‘universal’ protocol and leaf tissue, while in the remaining species the use of alternative tissues (such as petioles or flowers) and/or protocol modifications were needed (targeting composition of buffers, duration of incubation or staining time or use of alternative buffers). A total of 384 species (35%) included polyploid cytotypes. Among them, 151 (14%) included diploid and polyploid cytotypes, 25 species (2%) being ploidy variable with multiple polyploid cytotypes, and 208 (19%) being polyploids with just one cytotype. The remaining 719 (65%) species were diploid only. As a community resource, we provide relative genome sizes and ploidy assignments of 1,328 cytotypes retrieved from 1,103 species along with methodological details (e.g. buffers, standards, analysed plant organs, histogram quality, and other remarks). We believe that this dataset will facilitate future research in particular species as well as in flora-wide investigations of ploidy level variation of the central-European flora in general. We are confident that novel cytotypes of many species will be discovered in other geographic areas, and we would be delighted if the present dataset could serve the botanical community for comparison.