Variation of the nuclear, subnuclear and chromosomal flavanol deposition in hemlock and rye

Nuclei of hemlock (Tsuga canadensis and Tsuga canadensis var. nana) were investigated for the presence of flavanols. Histochemical staining with p-dimethylaminocinnamaldehyde proved to be a highly valuable method yielding a bright blue flavanol coloration for nuclei. There was a significant variation in flavanol deposition ( 1) among nuclei, ( 2) at the subnuclear level and also ( 3) along the chromosomes during mitosis. The presence of flavanols in nucleoli could not be established probably because they were too small, measuring less than 1 mu m in diameter. In contrast to Tsuga, the cells and nuclei of rootlets from rye ( Secale cereale) were totally devoid of natural flavanols. However, externally added flavanols, catechin and epicatechin, were bound to the rye nuclei, while the rather large nucleoli failed to associate with the flavanols. The strong sink activity of nucleoplasm and chromosomes for flavanols in Tsuga and Secale indicates a process which is apparently widespread even in distantly related plant species. Variations in chromatin-associated flavanols could to some extent be induced by acetylation/deacetylation of histones, as confirmed in the present study by means of UV-VIS spectroscopic titrations of histone sulphate and chemically acetylated histone sulphate.