Anti-phosphorylated histone H2AThr120: a universal microscopic marker for centromeric chromatin of mono- and holocentric plant species.

Based on the analysis of 20 different monocot and eudicot species, we propose that the centromeric distribution of the phosphorylated histone H2AThr120 is evolutionary highly conserved across species with mono- and holocentric chromosomes. Therefore, antibodies recognizing the phosphorylated threonine 120 of the histone H2A can serve as a universal marker for the cytological detection of centromeres of mono- and holokinetic plant species. In addition, super resolution microscopy of signals specific to the centromere-specific histone H3 variant CENH3 and to H2AThr120ph revealed that these histone variants are incorporated into different nucleosomes, which form distinct, partly intermingled chromatin domains. This specific arrangement of both histone variants suggests different centromeric functions during the cell cycle.

Holocentric chromosomes of Luzula elegans are characterized by a longitudinal centromere groove, chromosome bending, and a terminal nucleolus organizer region.

The structure of holocentric chromosomes was analyzed in mitotic cells of Luzula elegans. Light and scanning electron microscopy observations provided evidence for the existence of a longitudinal groove along each sister chromatid. The centromere-specific histone H3 variant, CENH3, colocalized with this groove and with microtubule attachment sites. The terminal chromosomal regions were CENH3-negative. During metaphase to anaphase transition, L. elegans chromosomes typically curved to a sickle-like shape, a process that is likely to be influenced by the pulling forces of microtubules along the holocentric axis towards the corresponding microtubule organizing regions. A single pair of 45S rDNA sites, situated distal to Arabidopsis-telomere repeats, was observed at the terminal region of one chromosome pair. We suggest that the 45S rDNA position in distal centromere-free regions could be required to ensure chromosome stability.

B chromosomes of Puschkinia libanotica are characterized by a reduced level of euchromatic histone H3 methylation marks.

Epigenetic marks in the chromosome complement of the liliaceous plant Puschkinia libanotica were studied using immunofluorescence to detect histone modifications. In particular, a comparison was made between the euchromatic and heterochromatic components of supernumerary B chromosomes (Bs) relative to the A chromosomes of the basic diploid set, in order to provide further insights into the enigmatic properties of these 'genetically silent' Bs. No differences were found between A and B chromosomes for the heterochromatic dimethylation marks of histone H3 at lysine positions 9 and 27. However, distinctions between A and B chromosomes were revealed for the euchromatic di- and trimethylation marks of histone H3 at lysine 4. The results indicate that the less-transcriptionally active Bs are not marked by an enriched level of heterochromatic histone marks, but rather by a low level of euchromatin-associated histone modifications.