RAPD Markers for the Study of Genetic Diversity in Echium amoenum Ecotypes.

This chapter details a comprehensive, step-by-step RAPD protocol for assessing genetic diversity in 16 Iranian Echium amoenum ecotypes, a medicinal plant. The method uses STE (Sucrose-Tris-EDTA) buffer to extract DNA from leaf tissue, followed by spectrophotometric and electrophoretic quality control. PCR amplification with 20 decamer primers yields 248 polymorphic bands (80% polymorphism), separated by agarose gel electrophoresis. NTSYS-pc analysis, using the Dice coefficient, UPGMA clustering, and principal component analysis, reveals geographic-climatic groupings. This approach is applicable not only to E. amoenum but also to other less-studied medicinal plants.