Genetic and Phytochemical Evaluation of Tissue Culture-derived Plants in Fennel (Foeniculum vulgar) under Different Concentrations of Growth Regulators

The genetic diversity among plants derived from tissue culture is called somaclonal variation, which provides a valuable source of genetic variation for the improvement of medicinal plants. The present study was conducted to investigate the efficiency of molecular markers in detection of somaclonal variation and to assess the importance of DNA methylation in occurrence of genomic changes. The genomic DNA of a normal plant and eight abnormal regenerated plants from calluses cultured in different conditions were extracted using modified Delaporta method. The AFLP procedure was performed with application of two different double digestion methods using restriction enzymes. The digested fragments were ligated to appropriate adaptors and amplification was carried out using appropriate primers. Also percentage and component of essential oil were indicated by GC/MS analysis. Analysis of banding patterns showed high differences in amount of polymorphism detected between two different double digestion methods. According to the results of cluster analysis based on the Jaccard's similarity coefficient, all tested plants divided into two main group. While the first group contained only normal sample, other abnormal samples were placed in the second group. Phytochemical analysis showed that the important secondary metabolites such as Limonene, Fenchone, Estragole, Anethole didn’t produce in invitro culture condition. In contrast some metabolites like Cineol, Terpineol, 2,4 Decadienal produce just in invitro culture. The results indicated that the used method has the potential to be used for assessment of somaclonal variations in regenerated plants. Additionally, considering characters of served enzymes in this study, phenotypic variations in abnormal plants that are resulted from somaclonal variation can be related to genome methylation.