| 基于ITS2序列及二级结构的维吾尔族药材孜然及其混伪品鉴别 |
| 投稿时间：2022-11-20 点此下载全文
|
| 引用本文：地力努尔·吐尔逊江,程波,何江.基于ITS2序列及二级结构的维吾尔族药材孜然及其混伪品鉴别[J].中国现代中药,2024,26(2):328-333 |
| DOI：10.13313/j.issn.1673-4890.20221120001 |
| 摘要点击次数: 255 |
| 全文下载次数: 0 |
|
|
| 基金项目:新疆维吾尔自治区公益性科研院所基本科研业务费项目（KY2021098）；国家中医药管理局青年岐黄学者项目 |
|
| 中文摘要:目的 应用DNA条形码技术，建立维吾尔族药材孜然及其混伪品小茴香、芫荽子的分子鉴别方法。方法 提取38批孜然及其混伪品的基因组总DNA，聚合酶链式反应（PCR）扩增内转录间隔区2（ITS2）序列并双向测序，测序结果经CodonCode Aligner软件进行序列拼接，利用MEGA软件分析序列特征，计算种内、属间遗传距离，构建邻接（neighbor-joining，NJ）系统聚类树，预测其ITS2二级结构。结果 孜然药材ITS2序列中主体单倍型为A1，序列长度均为226 bp，鸟嘌呤（G）+胞嘧啶（C）占比为47.79%~48.23%；小茴香ITS2序列中，主体单倍型为B1，序列长度均为224~227 bp，G+C占比为53.30%~55.36%；芫荽子ITS2序列中，主体单倍型为C1，序列长度均为220~227 bp，G+C占比为56.82%~56.83%。孜然的种内遗传距离明显小于属间遗传距离。二级结构表明，孜然及其混伪品螺旋区的茎环大小、数目、位置及螺旋角度均有明显差异。结论 ITS2序列作为DNA条形码能稳定、准确地鉴别孜然及其混伪品药材，为保障孜然临床安全用药提供了有效技术手段。 |
| 中文关键词:孜然 内转录间隔区2 二级结构 分子鉴别 |
| |
| Identification of Uighur Medicine Cuminum cyminum L. and Its Adulterants Based on ITS2 Sequences and Secondary Structure Analysis |
|
|
| Abstract:Objective Using DNA barcode technique, a molecular method was established to identify Cuminum cyminum L. and its adulterants, fennel and coriander seeds.Methods The total genomic DNA of 38 batches of Uygur medicine Cuminum cyminum L. and its adulterants was extracted, and the sequences of internal transcribed spacer region 2 (ITS2) were amplified by polymerase chain reaction (PCR) and sequenced in both directions. The sequencing results were assembled using CodonCode Aligner software, and the sequence characteristics were analyzed using MEGA software. Intraspecific and intergeneric genetic distances were calculated, and a neighbor-joining (NJ) phylogenetic tree was constructed to predict the secondary structure of ITS2.Results In the ITS2 sequence of Cuminum cyminum L., the main haplotype was A1, the sequence length was 226 bp, and the guanine (G) cytosine (C) content was 47.79%-48.23%. In the ITS2 sequence of Foeniculum vulgare Mill, the main haplotype was B1, the sequence length was 224-227 bp, and the GC content was 53.30%-55.36%. In the ITS2 sequence of Coriandrum sativum L., the main haplotype was C1, the sequence length was 220-227 bp, and the GC content was 56.82%-56.83%. The intraspecific genetic distance of Cuminum cyminum L. was significantly smaller than the interspecific genetic distance. The secondary structure analysis showed that there were significant differences in the size, number, position and helix angle of stem loops in the helical region between Cuminum cyminum L. and its adulterants.Conclusion As a DNA barcode, ITS2 sequence can stably and accurately distinguish Cuminum cyminum L. from its adulterants, providing a new and effective technical means for ensuring the clinical safety of cumin. |
| keywords:Cuminum cyminum L. ITS2 secondary structure molecular identification |
|
查看全文
查看/发表评论 下载PDF阅读器 |
|
|
|
