Liao Baosheng, Chen Xiaochen, Han Jianping, Dan Yang, Wang Lili, Jiao Wenjing, Song Jingyuan, Chen Shilin
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100193 People's Republic of China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700 People's Republic of China.
Chin Med. 2015 Aug 19;10:22. doi: 10.1186/s13020-015-0056-7. eCollection 2015.
DNA barcoding can be used to authenticate Ganoderma species for safe use. This study aims to identify commercial products containing Ganoderma using DNA barcoding.
We used 63 internal transcribed spacer (ITS) 2 sequences of Ganoderma species from 33 newly-sequenced wild samples, crude drugs, mycelia, spores, and authentic extracts and spore oils collected from various locations and markets, as well as 30 sequences from GenBank. Sequences were assembled and aligned using CodonCode Aligner V3.71. Intra- and inter-specific distances were estimated by MEGA 6.0, and phylogeny reconstruction was based on the K2P model. SNP(s) and RNA secondary structure of ITS2 were analyzed and compared among closely related Ganoderma species.
G. lucidum cultivated in China was different from those cultivated in Europe. "Chizhi" (G. lucidum) and "Zizhi" (G. sinense) were clustered into two clades that were separated from the other Ganoderma species. The fruiting bodies and commercial products of G. lucidum and G. sinense were successfully distinguished from those of other Ganoderma species by comparing the ITS2 sequences and RNA secondary structures.
The DNA barcoding method is applicable to the authentication of commercial products containing Ganoderma species.
DNA条形码技术可用于鉴定灵芝物种，以确保其安全使用。本研究旨在利用DNA条形码技术鉴定含灵芝的商业产品。
我们使用了63条灵芝物种的内部转录间隔区（ITS）2序列，这些序列来自33个新测序的野生样本、中药材、菌丝体、孢子，以及从不同地点和市场收集的正品提取物和孢子油，另外还使用了GenBank中的30条序列。使用CodonCode Aligner V3.71对序列进行组装和比对。通过MEGA 6.0估计种内和种间距离，并基于K2P模型进行系统发育重建。对亲缘关系密切的灵芝物种的ITS2的单核苷酸多态性（SNP）和RNA二级结构进行分析和比较。
中国栽培的灵芝与欧洲栽培的灵芝不同。“赤芝”（灵芝）和“紫芝”（紫芝）聚为两个分支，与其他灵芝物种分开。通过比较ITS2序列和RNA二级结构，成功区分了灵芝和紫芝的子实体及商业产品与其他灵芝物种的子实体及商业产品。
DNA条形码技术适用于含灵芝物种的商业产品的鉴定。
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