The effect of subculture on somaclonal variation in callus culture of Hyoscyamus niger using ISSR markers

To evaluate the genetic variation in an in vitro culture of two samples of Hyoscyamus niger callus including new (one-month-old) and old (one-year-old) calli versus fresh leaves of the plant, the Inter Simple Sequence Repeat (ISSR) marker was used. Using 10 primers, 6.49% polymorphism was observed in the three mentioned samples. Analysis of Molecular Variance (AMOVA) revealed 68% genetic variation among samples. Jaccard and Dice coefficients (similarity matrix) were also used by UPGMA and NJ methods. The best dendrogram to show genetic diversity (and distance or proximity of individuals) was known as the Jaccard similarity coefficient. The results of cluster analysis grouped the samples into two different groups. This classification was also confirmed by principal component analysis (PCoA). The first major group consisted of old calli and the second involved new calli and plant leaves. The result showed that the frequency of somaclonal variation in H. niger calli tended to increase with increasing the number of subculture times.