| 诃子基原物种及其混伪品的形态鉴别与分子鉴定 |
| 投稿时间：2024-02-22 点此下载全文
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| 引用本文：薛旭,武晓丽,丁自勉,奎学华,张娜娜,吴斌,邹忠梅.诃子基原物种及其混伪品的形态鉴别与分子鉴定[J].中国现代中药,2024,26(5):780-787 |
| DOI：10.13313/j.issn.1673-4890.20240222001 |
| 摘要点击次数: 157 |
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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 |
| 薛旭 |
XUE Xu |
中国医学科学院 北京协和医学院 药用植物研究所 濒危药材繁育国家工程实验室，北京 100193 |
National Engineering Laboratory of Endangered Medicinal Materials Breeding, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 武晓丽 |
WU Xiao-li |
中国医学科学院 北京协和医学院 药用植物研究所 濒危药材繁育国家工程实验室，北京 100193 |
National Engineering Laboratory of Endangered Medicinal Materials Breeding, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 丁自勉 |
DING Zi-mian |
中国医学科学院 北京协和医学院 药用植物研究所 濒危药材繁育国家工程实验室，北京 100193 |
National Engineering Laboratory of Endangered Medicinal Materials Breeding, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 奎学华 |
KUI Xue-hua |
云南省临沧市永德县林业调查规划设计队，云南 临沧 677600 |
Forestry Survey, Planning and Design Team of Yongde County, Lincang, Yunnan Province, Lincang 677600, China |
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| 张娜娜 |
ZHANG Na-na |
中国医学科学院 北京协和医学院 药用植物研究所 濒危药材繁育国家工程实验室，北京 100193 |
National Engineering Laboratory of Endangered Medicinal Materials Breeding, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 吴斌* |
WU Bin |
中国医学科学院 北京协和医学院 药用植物研究所 濒危药材繁育国家工程实验室，北京 100193 |
National Engineering Laboratory of Endangered Medicinal Materials Breeding, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 邹忠梅* |
ZOU Zhong-mei |
中国医学科学院 北京协和医学院 药用植物研究所 濒危药材繁育国家工程实验室，北京 100193 |
National Engineering Laboratory of Endangered Medicinal Materials Breeding, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 基金项目:中国医学科学院医学与健康科技创新工程项目（2022-I2M-1-017） |
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| 中文摘要:目的 建立能够准确鉴别诃子、绒毛诃子及其混伪品的分子鉴定方法。方法 对诃子、绒毛诃子及其混伪品的新鲜果实进行烘干，观察其干燥前后的形态；提取样品DNA，采用DNA条形码内转录间隔区2（ITS2）序列和psbA-trnH序列进行聚合酶链式反应扩增并测序，采用DANMAN 7、MAGE 6软件对测序结果进行序列比对，计算Kimura 2-paramete遗传距离，并构建邻接法系统发育树。结果 形态观察发现，诃子、绒毛诃子果实干燥前后与混伪品有较大的差异，可以通过形态差异进行鉴别；DNA条形码序列表明，诃子和绒毛诃子ITS2序列基本一致，仅发现4个单核苷酸多态性位点，但不能由此对两者进行区分；诃子和绒毛诃子的psbA-trnH序列存在2处明显差异，可对两者进行准确区分；绒毛诃子中存在2种类型的单倍型，其明显差异在于是否在psbA-trnH序列上存在1段16 bp的串联重复序列；混伪品与诃子、绒毛诃子之间具有较大的psbA-trnH序列差异，可直接在美国国家生物技术信息中心比对进行区分；对市场上流通的诃子炮制品进行鉴定发现，其多为2种单倍型绒毛诃子，且存在毛诃子混伪的情况。结论 利用psbA-trnH序列可准确鉴别诃子、绒毛诃子及其混伪品，发现绒毛诃子中存在2种类型的单倍型。 |
| 中文关键词:诃子 绒毛诃子 形态鉴别 DNA条形码 内转录间隔区2 psbA-trnH序列 分子鉴定 |
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| Morphological and Molecular Identification of Original Species of Chebulae Fructus from Adulterants |
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| Abstract:Objective To develop a precise molecular identification method for distinguishing Chebulae Fructus (Terminalia chebula), T. chebula var. tomentella, and their adulterants.Method The fresh fruits of T. chebula, T. chebula var. tomentella, and their adulterants were dried, and their morphological characteristics were observed before and after drying. Sample DNA was extracted, amplified, and sequenced using DNA barcode ITS2 and psbA-trnH sequences via polymerase chain reaction. The sequencing results were aligned using DANMAN 7 and MAGE 6 software to calculate Kimura 2-parameter genetic distance and create a neighbor-joining (NJ) phylogenetic tree.Results Morphological observations revealed significant differences between the fruits of T. chebula, T. chebula var. tomentella, and their adulterants before and after drying, which could be identified based on morphological distinctions. The DNA barcode sequence showed that the ITS2 sequences of T. chebula and T. chebula var. tomentella were largely similar, with only four single nucleotide polymorphism sites, making them indistinguishable based on this sequence. However, there were two distinct differences in the psbA-trnH sequences between T. chebula and T. chebula var. tomentella, allowing accurate differentiation between these two species. Two haplotypes were identified in T. chebula var. tomentella, with a significant variation related to the presence of a 16 bp tandem repeat sequence in the psbA-trnH sequence. Adulterants could be differentiated from T. chebula, T. chebula var. tomentella based on significant differences in the psbA-trnH sequence and confirmed through the National Center for Biotechnology Information. Identification of processed T. chebula in the market revealed that most samples were two haplotypes of T. chebula var. tomentella, with instances of adulterant T. bellirica.Conclusion The psbA-trnH sequence can be applied to accurately identifying T. chebula, T. chebula var. tomentella, and their adulterants, discovering two haplotypes in T. chebula var. tomentella. |
| keywords:Terminalia chebula Retz. Terminalia chebula Retz. var. tomentella Kurt. morphological identification DNA barcode ITS2 psbA-trnH sequence molecular identification |
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