| 引用本文:张嘉丽1,3，黄宇航1，宋明2，任阳阳1，张梦婷1，刘霞1，孙伟3，陈士林1,3.基于ITS序列鉴别真菌类药材马勃及其混伪品[J].世界中医药,2016,(05):. |
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| 基于ITS序列鉴别真菌类药材马勃及其混伪品 |
| Identification of Lasiosphaera Calvatia and Its Adulterants Using ITS Barcode |
| 投稿时间：2016-04-12 |
| DOI：10.3969/j.issn.1673-7202.2016.05.006 |
| 中文关键词: 马勃 混伪品 ITS 条形码 |
| English Keywords:Lasiosphaera Calvatia,ITS,adulterants,Barcoding |
| 基金项目:国家重大科技专项子课题“中药新药安全性检测技术与标准研究”——中药原料药混伪品分子基因鉴定技术与标准（编号：2014ZX09304-307-001-020） |
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| 中文摘要: |
| 目的:运用ITS序列鉴别真菌类药材马勃Lasiosphaera calvatia及其混伪品。方法：收集来自重庆、北京等9个地方的21份脱皮马勃、大马勃、紫色马勃样品，提取基因组DNA，经PCR扩增后双向测序。同时从GenBank上下载常见马勃药材混伪品的序列。将所有序列进行剪切校准拼接后分析，基于K2P（Kimura 2-Parameter）模型计算马勃及其混伪品的种内和种间遗传距离，并构建Neighbor-Joining(NJ)系统聚类树。结果：大马勃的ITS序列种内最大K2P遗传距离为0；脱皮马勃的ITS序列种内最大K2P遗传距离为0.003；紫色马勃的ITS序列种内最大遗传距离为0.003。大马勃与混伪品种间最小K2P遗传距离为0.019，脱皮马勃与混伪品种间最小K2P遗传距离为0.031，紫色马勃与混伪品种间最小K2P遗传距离为0.634。大马勃、脱皮马勃、紫色马勃的种内最大遗传距离均小于它们与混伪品的种间最小遗传距离。NJ树结果显示大马勃、紫色马勃、脱皮马勃各自聚为一支，表现出良好的单系性，能够与混伪品明显的区别开来。结论：基于ITS序列的条形码技术可以将马勃药材及其混伪品有效进行鉴别。 |
| English Summary: |
| ITS sequence was used as a barcode to identify Lasiosphaera Calvatia and its adulterants.Methods:We collected 21 samples of Lasiosphaera Calvatia and extracted their DNA，then amplified their ITS sequences.We also downloaded sequences of Lasiosphaera Calvatia’s adulterants from GenBank.Sequences were assembled and analysis their variable sites.The Kimura 2-Paramter(K2P)genetic distances and compute the Neighbor-joining(NJ)phylogenetic tree were computed.Results:The maximum intraspecific K2P genetic distances of the ITS sequence of Calvatia gigantea(Batsch ex Pers.)Lloyd is 0.The maximum intraspecific K2P genetic distances of the ITS sequence of Lasiosphaera fenzlii Reich.is 0.003.The maximum intraspecific K2P genetic distances of the ITS sequence lengths of Calvatia lilacina(Mont.et Berk.)Lloyd is 0.003.The minimum interspecific K2P genetic distances of Calvatia gigantea and adulterants is 0.019，Lasiosphaera fenzlii and adulterants is 0.031，while Calvatia lilacina and adulterants is 0.634.The maximum intraspecific genetic distances of Calvatia gigantea、Lasiophaera fenzlii and Calvatia lilacina were all less than the minimum interspecific genetic distances of themselves and their adulterants.the NJ tree based on ITS sequence showed that Calvatia gigantea、Lasiosphaera fenzlii and Calvatia lilacina can gather into a branch sperately.Conclusion:ITS sequence was able to identify Lasiosphaera Calvatia from their adulterants,which is suitable in the identification of Lasiosphaera Calvatia and its adulterants. |
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