Cytogenetic stability of maize tissue cultures: a cell line pedigree analysis

Abstract
Cytogenetic instability in maize plants regenerated from tissue culture is a commonly observed phenomenon. In an attempt to understand the origin of this instability meiotic analyses were performed on 370 regenerated plants that were initiated from cell lines of 22 immature embryos of 9 maize genotypes. Cell lineage pedigrees were maintained on these cultures to record the familial relationship between regenerated plants. Overall, 12.4% of the 370 régénérants contained cytological aberrations. The largest category of aberrations involved chromosome breakage (translocations, deletions, and an inversion) between a heterochromatic knob or knob site and the centromere. Late replicating knob heterochromatin might have been responsible for the chromosome breakage events. Cytologically identical aberrations were observed in plants regenerated at various time points in the pedigreed cultures, indicating an early occurrence of the aberration during culture maintenance. Heterogeneity of cell karyotypes also was observed within embryo cell lines. Multiple independent aberrations occurred within a single embryo cell line. The pedigree of the cultures provided evidence that the sectoring of regenerated plants was due to a multicellular origin of meristematic areas. The frequency of cytogenetically abnormal plants recovered increased with culture age. The data presented are consistent with the age effect not being due to an increased mutation rate but due to mutational events that occurred throughout culture development with a subsequent maintenance and accumulation of aberrant cells over time.Key words: Zea mays, cytogenetic stability, tissue culture, somaclonal variation.