Interphase chromosome organization is an important factor linked with chromatin states, gene expression and cell cycle stages. Understanding chromosome organization in cereals with large genomes is still in its infancy. We developed means of exploring dynamic chromosome organization in the small grain temperate cereal model barley, and aims to explore the role of nuclear lamina in the establishment and maintenance of Rabl chromosome configuration.

(Obj1) Quantify the temporal and spatial relationships between centromeres (CENH3) and nuclear lamina (CRWN1, KAKU4) proteins during interphase in wild-type using time-lapse imaging (confocal microscopy) and ChromTrack3D tools for segmentation, tracking and measurements. (Obj2) Elucidate the role of the nuclear lamina in establishing the Rabl configuration by analysing the spatio-temporal dynamics of chromosome positioning in crwn1 and kaku barley mutants as in Obj 1. (Obj3) Quantification of centromere association with nuclear lamina and envelope prior kinetochore binding and during mitotic chromosome segregation using super-resolution imaging of fixed nuclei (3D STED) using STED-optimized dyes for chromosomes, membranes and microtubules.