- 學位論文
細辛對於牛樟芝產孢作用及菌絲生長之影響
Effect of Asarum heterotropoides F. Schmidt var. mandshuricumon sporulation and mycelial growth of Antrodia cinnamomea
摘要
藥用菇類為現在大家注目的趨勢，因為它們具有療效，特別是當作化學治療和免疫調節劑。牛樟芝是一種傳統的藥用真菌，只生長在台灣特有的牛樟樹（Cinnamomum kanehirae）的褐腐性真菌。但由於寄主植物受到限制且生長速度緩慢，所以利用人工栽培的方式將牛樟芝培養於樟木上，試圖將牛樟芝以量產的方式來提高產率是大家想達到的目標。細辛屬於馬兜鈴科植物，有文獻報告指出它可以促進牛樟芝(也稱為台灣牛樟芝)的生長。因此，本實驗的目的要觀察利用細辛在正常情況下對於牛樟芝的產孢和菌絲生長之影響，是否可縮短菌絲生長的時間。 此實驗利用人工培養的方式選用兩種常用菌株AC1CK及ACN，以麥芽抽取物內含細辛萃取物當培養基，再用顯微鏡觀察牛樟芝的型態變化及蛋白質合成時RNA表現量的變化。 我們用了五種的細辛部分分層做測試，分別是細辛酒萃物(Ashe)、細辛水萃分層(Ashe-W)、細辛氯仿萃分層(Ashe-C)、細辛氯仿-己烷萃分層(Ashe-CH)、細辛氯仿-甲醇萃分層(Ashe-CM)，實驗濃度為500, 1000, 2000, 5000, 8000和 10000 ppm。從實驗結果裡面發現不管是AC1CK 或AC-N只有在加入2000 ppm細辛氯仿-己烷萃分層時促進生長活性最好，可以促進牛樟芝99.28 -150.36 %的菌絲生長。在液態培養下也是加入2000 ppm細辛氯仿-己烷萃分層時最佳，所測得的菌絲乾種分別AC1CK為908 mg/100 ml、AC-N為929 mg/100 ml、而未加細辛萃取物乾種分別為393 mg/100 ml 和 484 mg/100 ml。細辛的兩種純化合物asarinin和sesamin之實驗中，加入兩種純化合物與未加入化合物之牛樟菌絲可促進約120%的牛樟菌產孢率。從此實驗結果發現細辛氯仿-己烷萃分層可以促進牛樟芝菌絲的生長，此結果顯示可以在實驗室的環境下利用培養的方式縮短牛樟芝的生長時間及控制產量。
並列摘要
Medicinal mushrooms have attracted much attention recently owing to their potent therapeutic activity, especially as chemotherapy and immunomodulatory agents. Antrodia cinnamomea is an endemic medicinal fungus that grows mainly inside the rotten trunk of Cinnamomum kanehirae in natural conditions in Taiwan. Due to the limited distribution of the host plant and the slow growth rate of the fungus, the mass production of the fungus through in vitro culture systems including on the artificial cultural camphor tree for pharmaceutical usage has been attempted. Asarum heterotropoides F. Schmidt var. mandshuricumon that was assigned to birthwort family Aristolochiaceae, have been reported to promote growth of A. cinnamomea, also named Taiwanofungus camphorates. Therefore, the purpose of this study is to determine the effect of A. heterotropoides (Ashe) on sporulation and mycelial growth of A. cinnamomea in vitro and to detect the shorten growth time and formation of fruiting body quality compared with A. cinnamomea in natural. For sporulation and mycelial growth tests, two different strains of A. cinnamomea: AC1CK and AC-N were used and cultured with the extract from A. heterotropoides (Ashe). The malt extract agar (MEA) and a series of dilution of Ashe were made to culture for A. cinnamomea spores. Five fractions of Ashe, including Ashe E, Ashe W, Ashe C, Ashe CH, Ashe CM with different concentration 500 ppm, 1000 ppm and 2000 ppm were tested in this study. The results indicated that strain AC1CK and strain AC-N grew fastest in 2000 ppm among tested concentration as compared with other dosage. Ashe CH was the best among the partial fraction of Ashe to promote mycelial growth of A. cinnamomea at 2000 ppm. For mycelial growth tests, the results revealed that the Ashe CH obviously promoted the mycelial growth of A. cinnamomea to 99.28 -150.36 % in dose dependent manner with control. For pH tests, the maximum yield of 908 mg/100 ml, and 929 mg/100 ml, occurred at pH 5.0 in PDB with 2000 ppm Ashe CH of both strains AC1CK and AC-N. Furthermore, pure compounds from Ashe CH, asarinin and sesamin were used at concentrate 500 ppm to detect the effect on spore germination. The results indicated that spore germination rate of A. cinnamomea could be promoted to 120% as compared with untreated control. Based on the above results, the reliable production of the medicinal fungus can promote by Ashe treatment under laboratory conditions.