Cytological and RAPD (Random Amplified Polymorphic DNA) Analyses of Somaclonal variation in Easter Lily (Lilium longiflorum Thunb.)

Abstract

Long-term maintenance of plant cells in tissue culture is known to induce somaclonal variation in regenerated plants. Here we examined somaclonal variation in regenerated plants of Easter lily (Lilium longiflorum Thunb.) using cytological and molecular analyses. Forty nine 10-mer and 12-mer arbitrary primers were used to amplify DNAs extracted from sixty plants regenerated after more than two years of callus culture originally derived from bulb tissues. Sixteen primers were found to produce 34 polymorphic bands in the regenerated plants, including both loss of parental bands and appearance of novel, non-parental bands. The presence of polymorphic fragments among the regenerants indicates that genomic alterations occurred during the long-term culture of the cells. Although no variation was observed from karyotype analysis, the present study revealed the high frequency of the occurrence of somaclonal mutations at the DNA level during the callus culture of Easter lily.