
中药材蟾皮及其混伪品的DNA条形码鉴定研究
Identification of the Skin of Bufo bufo gargarizans and Its Adulterants Based on DNA Barcoding
目的 利用COI序列对中药材蟾皮及其混伪品进行DNA条形码鉴定研究,为蟾皮药材的快速准确鉴定提供新的技术手段。方法 收集16个不同地区的蟾皮药材、基原动物及其混伪品总计6个种54份样本,提取总DNA,进行PCR扩增及双向测序,测序结果采用CodonCode Aligner V 4.2进行拼接校对,运用MEGA5.0进行比对分析,计算种内及种间Kimura-2-Parameter(K2P)遗传距离并构建系统发育树。结果 蟾皮两个基原物种的种内最大K2P遗传距离均远远小于其与混伪品的种间最小K2P遗传距离。NJ树图显示,蟾皮药材的两个基原物种及其混伪品分别聚为独立一支,得到了很好区分,并显示出良好的单系性。结论 应用COI序列作为DNA条形码能够准确有效地鉴别中药材蟾皮及其混伪品。
OBJECTIVE To provide new technique for quick and accurate identification of the skin of Bufo bufo gargarizans using DNA barcoding method based on the COI sequence. METHODS Fifty-four samples of the skin of Bufo bufo gargarizans and its four kinds of adulterants were collected from 16 different localities. The DNAs were extracted, the COI squences were amplified and sequenced bi-directionally. The obtained sequences were assembled using the CodonCode Aligner V 4.2. Then the genetic distances were accumulated using Kimura-2-Parameter(K2P)model, and the NJ tree was constructed by MEGA5.0. RESULTS The intra-specific genetic distances were much smaller than inter-specific ones between the skin of Bufo bufo gargarizans and its adulterants. Furthermore, the NJ tree showed that the skin of Bufo bufo gargarizans can be distinguished from its adulterants clearly. CONCLUSION The COI sequence can be used as the DNA barcode to identify the skin of Bufo bufo gargarizans and its adulterants with accuracy and efficiency.
蟾皮 / DNA条形码 / 鉴别 / COI序列 {{custom_keyword}} /
skin of Bufo bufo gargarizans / DNA barcoding / identification / COI sequence {{custom_keyword}} /
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国家重大科技专项子课题资助项目(2014ZX09304-307-001-020)
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