| 不同种源银柴胡及其伪品的ITS2序列分析与鉴别 |
| 投稿时间：2022-04-11 点此下载全文
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| 引用本文：李振凯,冯璐,杨燕,宋乐,王红,李彦青,高跳,彭励.不同种源银柴胡及其伪品的ITS2序列分析与鉴别[J].中国现代中药,2022,24(12):2335-2341 |
| DOI：10.13313/j.issn.1673-4890.20220411004 |
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| 作者中文名 | 作者英文名 | 单位中文名 | 单位英文名 | E-Mail |
| 李振凯 |
LI Zhen-kai |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 冯璐 |
FENG Lu |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 杨燕 |
YANG Yan |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 宋乐 |
SONG Le |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 王红 |
WANG Hong |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 李彦青 |
LI Yan-qing |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 高跳 |
GAO Tiao |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 彭励* |
PENG Li |
宁夏大学 生命科学学院，宁夏 银川 750021 |
School of Life Sciences, Ningxia University, Yinchuan 750021, China |
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| 基金项目:宁夏回族自治区自然科学基金项目（2021AAC03103）；吴忠市科技创新驱动计划项目；吴忠市同心县科技发展计划项目 |
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| 中文摘要:目的 采用基于内转录间隔区2（ITS2）序列的分子鉴定法分别对17份不同种源银柴胡和6种伪品基原植物进行分析和鉴别。方法 对植物样品进行DNA提取、扩增、测序，并检索GenBank数据库，获取17份不同种源银柴胡及6种伪品的ITS2序列。采用ITS2数据库进行注释，去除5.8S和28S片段，获得完整的ITS2序列，利用MEGA 8.0软件进行序列比对、计算K2P遗传距离、构建邻接（NJ）系统发育树，采用美国国家生物技术信息中心（NCBI）数据库和“中药材DNA条形码鉴定系统”进行BLAST分析和物种鉴定，并利用ITS2数据库预测二级结构。结果 17份银柴胡样品共得到3个单倍型ITS2序列（YCH1、YCH2和YCH3），经与银柴胡标准ITS2序列比对，发现YCH1和YCH2与标准序列一致，但YCH3有4处碱基缺失，表现出0.936的遗传距离，在系统发育树中被单独聚为一个分支，并具有不同的二级结构；6种伪品的ITS2序列在长度、鸟嘌呤+胞嘧啶占比、变异位点、遗传距离、NJ系统发育树和二级结构上均与银柴胡ITS2序列表现出明显的差异。结论 基于ITS2序列的分子鉴定方法可以有效鉴别银柴胡真伪，不同种源银柴胡的遗传物质可能存在一定差异。 |
| 中文关键词:银柴胡 内转录间隔区2 DNA条形码 种质资源 真伪鉴别 |
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| Identification of Stellariae Radix and Its Counterfeits from Different Provenances Based on ITS2 Sequences |
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| Abstract:Objective To ensure the correct provenance and construct the seed breeding base of Stellariae Radix from the source, we identified 17 samples of Stellariae Radix from different sources and the original plants of 4 counterfeits based on internal transcribed spacer 2 (ITS2) sequence.Methods The DNA samples were extracted, amplified, sequenced, and searched against GenBank to obtain the ITS sequences of 17 samples of Stellariae Radix and the original plants of 6 counterfeits. MEGA 8.0 was used for sequence alignment, calculation of Kimura′s two-parameter (K2P) genetic distance, and construction of neighbor-joining (NJ) phylogenetic tree. Blast analysis and species identification were conducted via the NCBI database and the DNA barcoding system for Chinese herbal medicines. The secondary structure was predicted using the ITS2 Database online.Results A total of 3 haplotypes (YCH1, YCH2, and YCH3) of ITS2 sequences were obtained from 17 Stellariae Radix samples. YCH1 and YCH2 were consistent with the standard ITS2 sequence of Stellariae Radix. YCH3 had 4 base deletions, showing a genetic distance of 0.936, which was clustered into a branch alone in the phylogenetic tree and had a unique secondary structure. The ITS2 sequences of the original plants of 6 counterfeits showcased significant differences in length, guanine+cytosine content, mutation site, genetic distance, position on the phylogenetic tree, and secondary structure compared with the standard ITS2 sequence of Stellariae Radix.Conclusion The ITS2 sequence-based molecular method can effectively identify the authenticity of Stellariae Radix, and there may be genetic differences among different provenances. |
| keywords:Stellariae Radix ITS2 DNA barcode germplasm resources identification of authenticity |
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