Behaviour of chromosomes in anaphase cells in embryogenic callus cultures of maize (Zea mays L.)

Mitotic anaphase cells of highly friable and embryogenic calluses which had been induced from immature embryos of two inbred lines of maize that have contrasting levels of heterochromatic knobs were analysed for the presence of abnormalities 3, 6, 9 and 12 months after the initiation of culture. A total of 500 typical anaphases was scored at each time point, and various aberrations, such as delay in the separation of sister chromatides, chromosome bridges (single, double and multiple) and chromosome fragments, were revealed to occur extensively in the cultures of both genotypes. Preparations after C-banding revealed that primary breakages often occurred inside knobs or at junction regions between the euchromatin and the heterochromatin of the knobs. Figures characterized by the delayed separation of sister chromatids, which originated preferentially at the knob level and was considered to be an initial event in the development of breakages, were observed at constant frequencies throughout the experiment. Increasing numbers of aberrant cells were detected with time, mainly due to the accumulation of cells with chromosome bridges and fragments. Several mitotic figures suggested the occurrence of breakagefusion-bridge cycles that were initiated by broken chromosomes. The overall frequencies of aberrant cells were similar for both genotypes, despite the differences in knob composition. However, callus cultures induced from the genotype having the higher level of knobs had more aberrant cells with abnormalities that involved several chromosomes, such as multiple bridges and multiple fragments.