Identification of medicinal plants using PCR-RFLP in Dasamula - an Ayurvedic drug

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DOI: 10.4103/2231-4040.197331

Dasamula, an herbal drug formulation consisting of roots of 10 plants, is used widely for the treatment of various diseases but no reliable procedure is available to identify the presence of different prescribed plant material in the drug. Adulteration is rampant in the process of drug preparation. Molecular taxonomy is a reliable and easier way to identify any dried and processed medicinal plants and its adulterants from medicine by using DNA markers. Six out of ten plants of Dasamula i.e. Desmodium giganicum, Aegle marmelos, Solanum xanthocarpum, Solanum indicum, Tribulus terresteris, Oroxylum indicum plants have been studied to establish their identification using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique. Internal transcribed spacer (ITS) region of these plants were PCR amplified by using primers ITS1 (F) and ITS4(R). The size of ITS region varied from 650 bp to 850 bp. RFLP was carried out by restriction digestion of amplified ITS region using BamHI, HindIII, MspI, MboI, EcoRI, EcoRV, HinfI, AluI. RFLP showed species-specific variation in the number and size of digested DNA fragments. MspI and EcoRV proved to be the best enzymes as it could generate unique pattern for all the six plants. Variations in the ITS region among the species can be used for conclusive identification of these plant species.

Keywords: DNA molecular markers, ITS-RFLP, Dasamula herbal drug formulation, Restriction enzymes, Adulteration, Authentication