Molecular method for the identification of Bulbus Fritillariae Cirrhosae

The origin identification of Bulbus Fritillariae Cirrhosae is essential for clinical safety and efficacy.Previously reported identification methods based on macroscopical，microscopical or chemical characteristics lack specificity，therefore，the molecular identification method is of significance.Based on the continous investigation on the identification of Bulbus Fritillariae Cirrhosae in our laboratory，a simple and feasible PCR-RFLP method was newly developed.The experimental procedure was described as follows: after the pretreatment of 20 mg dried bulbs of Fritillaria with 75% ethanol and sterilized ultrapure water，genomic DNA was extracted with the novel and universal plant genomic DNA extraction kit，then PCR amplification of ITS₁ regions and restriction enzyme digestion reaction was carried out successively.The resultant electrophoresis spectrum showed that ITS₁ regions of Bulbus Fritillariae Cirrhosae were recognized by restriction enzyme SmaⅠwith providing 2 distinct fragments between 100 bp and 200 bp，while other species could not be digested.Finally，12 batches of commercial Bulbus Fritillariae were correctly differentiated through the above-mentioned method.