Trisomy associated with loss of maturation capacity in a long-term embryogenic culture of Abies alba

Abstract

 Karyological studies were made on a 6-year-old embryogenic cell line of Abies alba. Embryogenic cells were obtained from a mature zygotic embryo cultivated on modified MCM-medium and subcultured every 3 weeks. Three years after induction, part of the cell line was transferred to media supplemented with 500 or 1000 mg l^-1 caseine hydrolysate and 500 mg l^-1 L-glutamine. Approximately 3 years after addition of the organic nitrogen to the medium, morphological changes such as malformation of the suspensor cells and a loss of maturation capacity occurred. Chromosome counts revealed that all cells cultivated on medium with organic nitrogen were trisomic. Fluorescent-banding methods and comparison with an euploid cell line showed that the additional chromosome belonged to the group of long, metacentric chromosomes of Abies alba without secondary constriction. Those cells cultured on medium not supplemented with caseine hydrolysate and L-glutamine retained a stable chromosome number of 2n=24. Both normal and deformed suspensor cells were observed. The maturation frequency was very low. The emergence of aneuploidy within one cell line could be the consequence of high selection pressure caused by the different culture conditions.