Identification of <italic style="font-style: italic">Panax ginseng </italic>and <italic style="font-style: italic">Panax quinquefolius </italic>by Mitochondrial <italic style="font-style: italic">Cox</italic> Ⅱ Intron

Feng-jiao XU; Xiao-ran TIAN; Ming-ge QI; An-qi CHEN; Hong-tao WANG

doi:10.13422/j.cnki.syfjx.20201511

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Identification of Panax ginseng and Panax quinquefolius by Mitochondrial Cox Ⅱ Intron

Pharmacy Fundamentals | 更新时间：2020-08-11

- Identification of Panax ginseng and Panax quinquefolius by Mitochondrial Cox Ⅱ Intron
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 13, Pages: 163-167(2020)
- 作者机构：
  
  烟台大学生命科学学院，山东烟台 264005
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20201511
  CLC： R22;R289;R2-031;R284.2
- Published Online：13 April 2020，
  
  Published：05 July 2020
- 稿件说明：
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徐风娇,田潇然,齐明阁等.利用线粒体,cox Ⅱ内含子对西洋参和人参的分子鉴别[J].中国实验方剂学杂志,2020,26(13):163-167.

XU Feng-jiao,TIAN Xiao-ran,QI Ming-ge,et al.,Identification of ,Panax ginseng and ,Panax quinquefolius by Mitochondrial ,Cox,Ⅱ, Intron[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(13):163-167.
徐风娇,田潇然,齐明阁等.利用线粒体,cox Ⅱ内含子对西洋参和人参的分子鉴别[J].中国实验方剂学杂志,2020,26(13):163-167. DOI： 10.13422/j.cnki.syfjx.20201511.

XU Feng-jiao,TIAN Xiao-ran,QI Ming-ge,et al.,Identification of ,Panax ginseng and ,Panax quinquefolius by Mitochondrial ,Cox,Ⅱ, Intron[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(13):163-167. DOI： 10.13422/j.cnki.syfjx.20201511.

摘要

目的

基于西洋参和人参在线粒体细胞色素C氧化酶亚基Ⅱ（

cox

Ⅱ）的内含子对西洋参和人参进行鉴别，为人参类药材提供简单准确的分子检测手段。

方法

利用通用引物对人参和西洋参线粒体

cox

Ⅱ基因的内含子进行聚合酶链式反应（PCR）扩增，将扩增后的序列进行双向测序，通过序列比对挖掘人参和西洋参的插入/缺失序列。根据人参和西洋参的特异性插入序列分别设计两者的品种特异性引物，建立鉴别人参和西洋参的多重PCR体系，并确定该多重PCR体系的检测限。

结果

在线粒体

cox

Ⅱ的内含子中发掘了人参和西洋参的插入/缺失序列。在多重PCR体系下，人参产生了729 bp 的条带，西洋参产生了141 bp的条带，人参和西洋参的混合品则分别产生了729 bp和141 bp的条带。该多重PCR体系可以检测人参中0.1%的西洋参掺入，检测限低至0.001 ng。

结论

该方法建立的多重PCR体系既不需要对反应体系进行优化，也不需要引入额外的碱基错配，可以对不同来源的人参和西洋参进行准确的鉴定，为人参和西洋参植物来源的鉴定提供了新的分子标记方法。

Abstract

Objective

To develop a simple and accurate method for molecular authentication of

Panax ginseng

and

P. quinquefolius

Method

The mitochondrial

cox

Ⅱ sequences of

P. ginseng

and

P. quinquefolius

were amplified by polymerase chain reaction（PCR）with universal primers. PCR products of the two species were sequenced in both directions

and sequence alignments were conducted for intron length polymorphisms exploitation. Multiplex PCR was established for the identification of

P. ginseng

and

P. quinquefolius

with their specific primers，which were designed respectively based on their insertion sequences. And the limit of detection of the multiplex PCR was also determined.

Result

The insertion/deletion sequences were exploited in mitochondrial

cox

Ⅱ. Under the established multiplex PCR assay，

P. ginseng

generated a 729 bp specific band

while

P. quinquefolius

yielded a 141 bp specific amplicon，and the mixture of the two species yielded both 729 bp and 141 bp fragments. The established multiplex PCR assay could detect 0.1% of intentional adulteration of

P. quinquefolius

into

P. ginseng

with down to 0.001 ng of genomic DNA.

Conclusion

The established multiplex PCR assay can accurately identify

P. ginseng

and

P. quinquefolius

from different sources

without the optimization of reaction system and the introduction of additional mismatches，so as to provide a new molecular marker method for identifying botanical origin of

P. ginseng

and

P. quinquefolius.

关键词

Keywords

references

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Identification of Panax ginseng and Panax quinquefolius by Mitochondrial Cox Ⅱ Intron

DOI：10.13422/j.cnki.syfjx.20201511

摘要

Abstract

关键词

Keywords

references