Assessment of somaclonal variation of regenerated Ducrosia anethifolia plants using AFLP markers.

Ducrosia anethifolia is a medicinal plant of Apiaceae family. The plant contains active constituents with antimicrobial activity which makes it as a valuable crude drug for use in pharmaceutical and food industries. Besides others, in vitro production of secondary products is an appropriate way of production of theses group of metabolites. In these cases, many factors such as number of subcultures, explant, types of culture media and their constituent's especially exogenous hormones may lead to genetically and epigenetically changes along with DNA methylation which can be exploited as a desirable phenomenon. The present study was done to investigate somaclonal variation in regenerated D. anethifolia) plants and to study importance of DNA methylation in occurrence of somaclonal variation. To do this, eight abnormal plants regenerated from different explants calli along as well as a normal plant were selected and their genomic DNA were extracted using modified Delaporta method. AFLP analysis of selected plants was performed using two different digestion systems. Digested DNA fragments were ligated to adapters and amplification was done using suitable primers. Using two different digestion systems, considerable polymorphism was observed. Considering obtained polymorphism and differences of used enzymes to methylation, it can be concluded that observed variations in regenerated plants are resulted from genome methylation. Additionally, the applied method has the potential to be used for assessment of somaclonal variations for in vitro regenerated plants.