Leaves and Seeds as Materials for Flow Cytometric Estimation of the Genome Size of 11 Rosaceae Woody Species Containing DNA-Staining Inhibitors

Abstract

The presence of some secondary metabolites in the cell cytosol can cause a stoichiometric error in the flow cytometric estimation of nuclear DNA content. There is no fully reliable method to completely eliminate the effect of these compounds on nuclei fluorescence, and therefore using plant organs/parts free of staining inhibitors is recommended. Eleven species of Rosaceae with high concentrations of propidium-iodide-staining inhibitors were studied to check the possibility of using seeds instead of leaves for genome size estimation. Despite optimizing the concentration and composition of antioxidants in nuclei-isolation buffer for each species, the effect of cytosolic compounds present in the leaves could not be avoided entirely. None of the seeds of the studied species contained inhibitors, and they produced histograms of good quality. The genome size of the studied species ranged from 1.15 to 3.17 pg/2C; for 10 species the DNA content was estimated for the first time.