Identification of an EcoRI restriction site for a rapid and precise determination of β-asarone-free Acorus calamus cytotypes

The aim of this work was to identify a diploid β-asarone-free Acorus calamus by using chemical and molecular approaches. A. calamus was analyzed by gas chromatography-mass spectrometry (GC-MS) and comparison of the 700 bp sequence of a 5S-rRNA gene spacer region was performed. By aligning the isolated nucleotide sequences of varieties of A. calamus chemotypes present in Genbank, sequence diversities were found in the spacer region. The PCR products were digested by using EcoRI. The restriction profile of the spacer domain resulted different for the two cytotypes.