Authentication of Herbal Medicinal Plant-Boerhavia diffusa L. Using PCR-RFLP

Herbal drugs are integrated part of both modern and traditional systems of medicine and are frequently found to be adulterated with other drugs of same morphological features, thereby requires scientific methods for their authentication and standardization. Various methods like chemo-profiling using chromatographic and spectroscopic techniques are used for the purpose but have their limitations because of the variations in the chemistry of the metabolites due to the age of the plants and their varied geographical distribution. DNA-based techniques have been widely used as a method of authentication for such types of herbal drugs. In the present study, using a DNA based technique known as PCR-RFLP, an attempt has been made to authenticate one such herbal drug Boerhavia diffusa which gets adulterated with Trianthema portulacastrum and Trianthema monogyna. The PCR amplified product of ribosomal ITS gene when subjected to restriction digestion with five different restriction enzymes viz. MspI, HinfI, MboI, EcoRI and EcoRV showed varied banding patterns for Boerhavia diffusa and adulterated plants. Of the five enzymes MspI, HinfI and MboI could be used for authentication of B. diffusa as they gave unique patterns for this plant and were different from that of adulterants. Rest of the enzymes either could not digest the ITS product or was not showing sufficient polymorphism to assign it to a particular genotype. Our study has established ITS-RFLP based markers for the authentication of B. diffusa, T. portulacastrum and T. monogyna with enzymes MspI, HinfI and MboI.