Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method for distinguishing honeysuckle and lonicerae flos

Abstract

The invention belongs to the technical field of distinguishing traditional Chinese medicinal materials, and in particular relates to a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method for distinguishing honeysuckle and lonicerae flos. The feature DNA base sequence of the honeysuckle is 5'-Y GGCCR-3' and can be identified and cut by restriction enzyme CfrI or EaeI. The distinguishing process comprises the following steps of: extracting medicinal material DNA; performing PCR amplification by a pair of primers; digesting a PCR product by the restriction enzyme CfrI or EaeI; performing agarose gel electrophoresis analysis on the result; and comparing the result with a PCR-RFLP feature result of the lonicerae flos, and judging whether the test piece is the honeysuckle or the lonicerae flos. The quick, convenient and reliable PCR-RFLP method is realized according to the difference between the DNA sequences of the honeysuckle and the lonicerae flos; and the PCR-RFLP method has an important significance for guaranteeing the quality of a honeysuckle medicinal material, cracking down the ill behavior that sellers sell the lonicerae flos as the honeysuckle and protecting the rights and interests of customers.

Description

technical field

[0001] The invention belongs to the technical field of germplasm identification of traditional Chinese medicinal materials, and in particular relates to a PCR-RFLP method for identifying honeysuckle and mountain silver flowers. Background technique

[0002] The 2010 edition of "The Pharmacopoeia of the People's Republic of China" records that honeysuckle is the plant Lonicera Lonicera ( Lonicera japonica Thunb.) dry flower buds or with first-opened flowers, mountain silver flower is the Lonicera chinensis Lonicerae Lonicerae ( Lonicera macranthoides Hand. -Mazz.), red gland honeysuckle ( Lonicera hypoglauca Miq.), South China honeysuckle ( Lonicera confusa DC.) and Tawny Honeysuckle ( Lonicera fulvotomentosa Hsu et S.C.Cheng.) dry buds or with first flowers. Honeysuckle and mountain silver flower are very similar in plant morphology and medicinal material shape, so it is difficult to distinguish them. However, there are obvious differences in the e...

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