长梗黄精的SCAR分子鉴别

叶碧欢, 陈友吾, 胡传久, 宋其岩, 廖荣俊, 翁永发, 杜国坚, 李海波

中国药学杂志 ›› 2019, Vol. 54 ›› Issue (20) : 1647-1652.

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中国药学杂志 ›› 2019, Vol. 54 ›› Issue (20) : 1647-1652. DOI: 10.11669/cpj.2019.20.003
论著

长梗黄精的SCAR分子鉴别

  • 叶碧欢1, 陈友吾1, 胡传久1, 宋其岩1, 廖荣俊2, 翁永发2, 杜国坚1, 李海波1*
作者信息 +

SCAR Molecular Identification of Polygonatum filipe

  • YE Bi-huan1, CHEN You-wu1, HU Chuan-jiu1, SONG Qi-yan1, LIAO Rong-jun2, WENG Yong-fa2, DU Guo-jian1, LI Hai-bo1*
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摘要

目的 建立一种快速鉴别长梗黄精种的分子技术手段。方法 基于简单序列重复区间扩增多态性(ISSR)和相关序列扩增多态性(SRAP)分子标记对多花黄精与长梗黄精的基因组DNA进行种间的多态性分析,获得ISSR和SRAP-PCR差异片段,经测序设计种特异性的特征性片段扩增区域(SCAR)引物,用于二种黄精属植物的准确快速鉴别。结果 3对SCAR引物在各自最适退火温度下,分别只从长梗黄精中扩增出了150、354和518 bp的特异性片段,而多花黄精不扩增任何片段,实验操作便捷,结果重复性好。通过对市场上8份黄精块茎材料的鉴定,SCAR分子鉴别技术的可靠性进一步得以验证。结论 本实验开发的SCAR分子技术可用于长梗黄精种的辅助鉴别。

Abstract

OBJECTIVE To establish a rapid molecular identification method for Polygonatum filipe species. METHODS Polymorphism analysis on DNA of P. filipe and P. cyrtonema was performed by using inter simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) molecular markers. Differential ISSR and SRAP bands between the two species were sequenced and species-specific sequence characterized amplified region (SCAR) primers were designed for the identification of P. filipe and P. cyrtonema. RESULTS Under respective optimal annealing temperature, three pairs of SCAR primers can specifically amplify three fragments of 150, 354 and 518 bp only from P. filipe, respectively, not from P. cyrtonema. The SCAR-PCR test was simple and convinent to operate, and reproducible. The molecular identification technology based on SCAR markers was further validated by testing 8 samples of Polygonatum tubes sold in market. CONCLUSION SCAR molecular technology developed in this study can be used for the assistant identification of P. filipe species.

关键词

长梗黄精 / 多花黄精 / 种间鉴别 / 分子标记

Key words

Polygonatum filipe / P. cyrtonema / species identification / molecular marker

引用本文

导出引用
叶碧欢, 陈友吾, 胡传久, 宋其岩, 廖荣俊, 翁永发, 杜国坚, 李海波. 长梗黄精的SCAR分子鉴别[J]. 中国药学杂志, 2019, 54(20): 1647-1652 https://doi.org/10.11669/cpj.2019.20.003
YE Bi-huan, CHEN You-wu, HU Chuan-jiu, SONG Qi-yan, LIAO Rong-jun, WENG Yong-fa, DU Guo-jian, LI Hai-bo. SCAR Molecular Identification of Polygonatum filipe[J]. Chinese Pharmaceutical Journal, 2019, 54(20): 1647-1652 https://doi.org/10.11669/cpj.2019.20.003
中图分类号: R282   

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基金

浙江省中央财政林业科技推广项目资助(2017-TS04);浙江省衢州市科技攻关项目资助(2018-K45)
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