Specific PCR Identification of Hibisci Cortex and Its Adulterants Based on DNA Signature Sequence Tags
10.13422/j.cnki.syfjx.20221013
- VernacularTitle:基于DSS标记特异性PCR鉴别冷背药材木槿皮基原植物及其混伪品
- Author:
Yanan LIU
1
;
Zhongyi HUA
2
;
Yuyang ZHAO
2
;
Yan JIN
2
;
Huangsheng PENG
1
;
Chao JIANG
2
;
Jingzhe PU
3
;
Yuan YUAN
1
Author Information
1. School of Pharmacy， Anhui University of Chinese Medicine， Hefei 230012， China
2. State Key Laboratory Breeding Base of Dao-Di Herbs， National Resource Center for Chinese Materia Medica， China Academy of Chinese Medical Sciences， Beijing 100700， China
3. Anhui Institute of Food and Drug Inspection， Hefei 230051， China
- Publication Type:Journal Article
- Keywords:
DNA signature sequence tag;
Hibisci Cortex;
adulterants;
allele-specific polymerase chain reaction（PCR）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(17):133-139
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveUncommon medicinal herbs are valuable medicinal resources， but their identification is a difficult problem in Chinese medicine due to their particularity and complexity. It is， therefore， urgent to establish a method for the identification of uncommon medicinal herbs. In this study， DNA signature sequence （DSS） tags were used to establish a specific polymerase chain reaction （PCR） identification method for Hibisci Cortex， the origin plant of Hibisci Cortex， and its adulterants. MethodThe candidate DSS tags were obtained from the chloroplast genome sequence analysis， and the DSS tags were verified by DNA sequencing. The specific identification primers for H. syriacus were designed based on the obtained reliable DSS tags. The PCR reaction conditions were optimized， and the tolerance and feasibility were investigated. ResultA DSS tag for identification of H. syriacus was obtained from the comparison of sequencing results of the amplified products with DSS， which revealed the distinguishing characteristics of Hibisci Cortex and its adulterants. A pair of specific primers for H. syriacus was designed according to the DSS tag. After PCR amplification and gel electrophoresis with the primers， a single bright band of about 270 bp was observed from H. syriacus， which did not appear in the four adulterants. ConclusionA DSS tag obtained in this study can be used to identify H. syriacus. The specific primers designed based on this DSS tag can accurately and simply identify the original plant of Hibisci Cortex and its adulterants， which provides a new method and idea for the molecular identification of genuine and counterfeit products of Hibisci Cortex.
