Objective

2

In recent years， with the sharp decline of wild resources in Arisaematis Rhizoma and Pinelliae Rhizoma and the immaturity of medicinal cultivation technology， their adulterants have appeared frequently in the market， and the main identifying characteristics have mostly disappeared in the circulation of medicinal materials. Therefore， there is an urgent need to establish a molecular identification method that can quickly and effectively identify the specificity of Arisaematis Rhizoma

and Pinelliae Rhizoma

.

Method

2

After comparison of the

rbc

L sequences of Arisaematis Rhizoma，Pinelliae Rhizoma， and their adulterants， the specific enzyme cleavage sites

Hae

Ⅲ and

Dra

Ⅰ of Arisaematis Rhizoma

and Pinelliae Rhizoma， respectively， were selected and identified by polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）. The main system conditions of PCR-RFLP reaction were established and optimized， and their durability and the ability to detect genuine， adulterants， and mixed counterfeits were investigated.

Result

2

The PCR-RFLP identification method of Arisaematis Rhizoma

and

Pinelliae Rhizoma

was established. After specific primer amplification， Arisaematis Rhizoma

and Pinelliae Rhizoma could be digested by

Hae

Ⅲ and

Dra

Ⅰ-restricted endonucleases respectively， at annealing temperature of 54 ℃， the number of cycles of 35， and the amount of DNA template of 3-30 ng， producing two fragments or small cut fragments with a single band between 100-250 bp， whereas the mixed counterfeits were not cleaved and both showed a band at 250 bp. The method is highly accurate in identifying adulterants and mixed counterfeits of Arisaematis Rhizoma or Pinelliae Rhizoma.

Conclusion

2

The PCR-RFLP method developed in this study allows for the rapid identification of Arisaematis Rhizoma and

Pinelliae Rhizoma.