Identification of Arisaematis Rhizoma<italic style="font-style: italic"> </italic>and<italic style="font-style: italic"> </italic>Pinelliae Rhizoma by<italic style="font-style: italic"> </italic>PCR-RFLP

XIAO Jiancai; YAN Binbin; YANG Jian; SUN Jiahui; WANG Tielin; WAN Xiufu; SUN Kai; JIANG Chao; ZHANG Yan

doi:10.13422/j.cnki.syfjx.20230116

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Identification of Arisaematis Rhizoma and Pinelliae Rhizoma by PCR-RFLP

更新时间：2023-02-15

- Identification of Arisaematis Rhizoma and Pinelliae Rhizoma by PCR-RFLP
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 6, Pages: 194-201(2023)
- 作者机构：
  
  中国中医科学院中药资源中心道地药材国家重点实验室培育基地，北京 100700
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230116
  CLC： R284.2;R285;R289;R22;R2-031;R33
- Published：20 March 2023，
  
  Published Online：16 December 2022，
  
  Received：20 August 2022，
- 稿件说明：
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肖建才,闫滨滨,杨健等.天南星、半夏的PCR-RFLP鉴别[J].中国实验方剂学杂志,2023,29(06):194-201.

XIAO Jiancai,YAN Binbin,YANG Jian,et al.Identification of Arisaematis RhizomaandPinelliae Rhizoma byPCR-RFLP[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(06):194-201.
肖建才,闫滨滨,杨健等.天南星、半夏的PCR-RFLP鉴别[J].中国实验方剂学杂志,2023,29(06):194-201. DOI： 10.13422/j.cnki.syfjx.20230116.

XIAO Jiancai,YAN Binbin,YANG Jian,et al.Identification of Arisaematis RhizomaandPinelliae Rhizoma byPCR-RFLP[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(06):194-201. DOI： 10.13422/j.cnki.syfjx.20230116.

摘要

目的

近年来，随着半夏和天南星的野生资源急剧下降、药材栽培技术不成熟等问题出现，市场的混伪品频繁出现，而其主要鉴别特征在药材流通中大多消失。因此，迫切需要建立一种可以快速、有效鉴别天南星和半夏特异性的分子鉴定方法。

方法

通过比对天南星、半夏及其混伪品的

rbc

L序列，分别选择了天南星和半夏的特异性酶切位点

Hae

Ⅲ和

Dra

Ⅰ，采用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）进行鉴定，建立并优化了PCR-RFLP反应主要体系条件，并对其耐用性和正品、掺杂品及混伪品的检出能力进行了考察。

结果

建立了天南星和半夏药材的PCR-RFLP鉴别方法，在退火温度为54 ℃、循环次数为35个，DNA模板量为3~30 ng时，天南星和半夏经过特异性引物扩增后，分别能被

Hae

Ⅲ和

Dra

Ⅰ限制性内切酶酶切，产生两片段或切开的小片段，并在100~250 bp出现单一条带，而混伪品无法酶切，且均在250 bp出现条带，该方法对天南星或半夏的掺杂品和混伪品具有高度准确的鉴别能力。

结论

该研究建立的PCR-RFLP方法可以快速鉴别天南星、半夏。

Abstract

Objective

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

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

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

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

Pinelliae Rhizoma.

关键词

天南星半夏聚合酶链反应-限制性片段长度多态性方法混伪品分子鉴定

Keywords

Arisaematis RhizomaPinelliae Rhizomapolymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）adulterantsmolecular identification

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Identification of Arisaematis Rhizoma and Pinelliae Rhizoma by PCR-RFLP

DOI：10.13422/j.cnki.syfjx.20230116

摘要

Abstract

关键词

Keywords

references