Establishment of a Rapid PCR Detection Method for ＂Antrodia salmonea＂ and ＂A. cinnamomea＂

牛樟芝（＂Antrodia cinnamomea＂）及香杉芝（＂A. ntrodia salmonea＂）均屬多孔菌科（Polyporaceae）真菌。牛樟芝是台灣特有的珍貴真菌，只生長在台灣特有的牛樟樹，貼生於樹幹中空之內面。香杉芝生長在已腐朽的香杉樹幹，以前被誤認為牛樟芝，於2004年才被認定為＂Antrodia＂屬之新種。這兩種真菌由於形態相似，通常以孢子形態、子實體、寄主偏好和交配型作為區隔。香杉芝常被不肖業者用作牛樟芝的替代物，但傳統鑑定方法上，此兩種菌種的鑑定需要專業知識及經驗，因此需要真菌鑑定專業人員參與，鑑定作業實屬不易。在這項研究中，我們測試了六組真菌引子對，並且解析了這兩真菌之間的序列，我們發現其中一引子對進行PCR增幅時，香杉芝比牛樟芝多出一段1.5 kb之核酸片段，我們並根據此片段另外設計了一組引子對Acl-F／Acl-R，增幅片段為219 bp，可針對香杉芝進行快速鑑定。本研究建立快速而準確的牛樟芝與香杉芝菌種鑑定方法，並建立快速鑑定流程，檢測靈敏度高，且鑑定時間僅需5小時，可提供牛樟芝與香杉芝之鑑定辨識，以及提供產業界及公務部門鑑定之依據。

關鍵字

牛樟芝；香杉芝；聚合酵素連鎖反應專一性鑑定

並列摘要

Both ＂Antrodia cinnamomea＂ and ＂A. salmonea＂ are fungi of the Polyporaceae. ＂Antrodia cinnamomea＂, which only grows on the inner side of the trunk of ＂Cinnamomum kanehirai＂ Hayata (Lauraceae), is a precious medicinal fungus; ＂A. salmonea＂ causes brown heart rot disease of ＂Cunninghamia konishii＂ Hayata (Cunninghamieae) in Taiwan. It is difficult to distinguish ＂A. cinnamomea＂ from ＂A. salmonea＂ because of their morphological similarities. ＂Antrodia salmonea＂ is commonly used as a counterfeit substitute for ＂A. cinnamomea＂ and sold by dishonest merchants. In 2004, these 2 fungi were first distinguished by the pore surface color of the basidiomata, host preferences, and mating types. However, accurate identification relies on professional training and experience. Moreconvenient and-persuasive methods are necessary for precisely identifying ＂A. cinnamomea＂. In this study, we applied 6 fungal primer pairs in polymerase chain reaction (PCR) assays, and analyzed sequences of amplified DNA fragments between ＂A. cinnamomea＂ and ＂A. salmonea＂. Results showed that one of these primer pairs could amplify a particular DNA fragment from ＂A. salmonea＂, which was approximately 1.5 kb longer than that from ＂A. cinnamomea＂. Based on this 1.5-kb difference in sequences, another primer pair named Acl-F/Acl-R was designed for the specific detection of ＂A. salmonea＂. A specific fragment, of 219 bp, was yielded only from A. salmonea, whereas no fragment was yielded from A. cinnamomea by the PCR assay. In this study, we established a rapid and accurate identification technique for A. salmonea, which will be helpful in rapidly differentiating ＂A. salmonea＂ from ＂A. cinnamomea＂. Additionally it also saves significant time as the entire procedure only takes about 1 h and has high sensitivity. This identification method can provide objective evidence for industry and public institutions.

並列關鍵字

Antrodia cinnamomea ； Antrodia salmonea ； specific PCR detection

國際替代計量

Establishment of a Rapid PCR Detection Method for ＂Antrodia salmonea＂ and ＂A. cinnamomea＂

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