北蟲草栽培鑑定與生物活性之研究

The culture, identification and bioactivity analysis of Cordyceps militaris

丘錦朋(Ching-Peng Chiu)

指導教授 ： 錢佑

中山醫學大學/醫學院/醫學應用微生物研究所/碩士(2011年)

摘要

冬蟲夏草(Cordyceps sinensis)，又稱作中華蟲草，自古以來就是很珍貴的藥用真菌，冬蟲夏草生長環境嚴苛因此取得不易。北蟲草(Cordyceps militaris)為麥角菌科(Clavicipitaceae)，蟲草屬的一種藥用真菌，其生活史與冬蟲夏草相同。在古典籍中有提到北蟲草的子實體及蟲體也可作為冬蟲夏草入藥，更有科學研究指出北蟲草和冬蟲夏草有很相似成份。在自然界中北蟲草亦是一種罕見的藥用真菌，但最大特點在於北蟲草可以用人工栽培的方式生產。本實驗室經過了多年努力，在培養基、溫度、溼度、光照項目經研究最佳條件，建立北蟲草子實體無菌人工量化栽培方法。本研究使用隨機擴增多態性DNA技術分析法(Random amplified polymorphic DNA，簡稱RAPD)來檢測北蟲草與中華蟲草的基因組序列，以5組中華蟲草基因鑑定常用的引子(Primer)來探討兩者異同之處。在這項研究中發現本實驗室栽培的北蟲草與標準品冬蟲夏草比較。此外，進一步使用不同的分配係數(Distribution coefficient)原理設計了萃取方法，研發出六種不同萃取物CM-1~CM-6。研究其化學成份、不同產率、成份組成以及其抗氧化相關活性進行研究。綜合以上結果，我們得到結論本實驗室量化生產北蟲草子實體並與中華蟲草基因序列進行比較，且經由不同分配係數萃取產生之粗萃取物，可以獲得良好的抗氧化活性。將進一步針對成份及其藥理活性進行研究。

關鍵字

冬蟲夏草 ； 北蟲草 ； RAPD ； NMR ； 抗氧化活性

並列摘要

Cordyceps sinensis is the most seldom and valuable herb used in Chinese medicine. Cordyceps sinensis are difficult to obtain because of harsh growth conditions in the natural environment. Cordyceps militaris belongs to Clavicipitaceae. Traditional Chinese medicine books described Cordyceps militaris as “fruiting bodies and caterpillar can be used as medicine for Cordyceps sinensis”. In addition, scientific studies have proved that Cordyceps militaris has ingredients similar to Cordyceps sinensis.
The wild Cordyceps militaris is a rare medicinal fungus with the specific advantage of being able to be produced by artificial culture. Our laboratory attempted diligently for many years to determine the optimal conditions of culture medium, temperature, humidity, and light source to establish a mass production method for Cordyceps militaris. This study defines genetic DNA sequences of Cordyceps militaris and Cordyceps sinensis using a Random Amplification of Polymorphic DNA (RAPD) technique, and analyzed the relationship between Cordyceps militaris and Cordyceps sinensis using five identification primers. The result in comparison of standard Cordyceps sinensis and culture Cordyceps militaris was discovered and revealed in this study. Moreover, extraction methods were designed following different the theory of solvent distribution coefficient. The design lead to six extracts CM-1~CM-6. The yields chemical compositions and their antioxidant effects were also studied. On the basis of results obtained, we conclude that: the genomic sequences of Cordyceps militaris optimally quantity-yielded are similar to those of Cordyceps sinensis. All of the extracts show positive antioxidant activity Cordyceps militaris is numerous potentially valuable to be developed, and the pharmacological activity of Cordyceps militaris is worthy of further investigation.

並列關鍵字

Cordyceps sinensis ； Cordyceps militaris ； RAPD ； NMR ； antioxidant activity

參考文獻

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