臺灣博碩士論文加值系統

本研究經由共培養使菌種間進行相互作用以誘導多樣的二次代謝產物，並對菌絲體施以LED燈光照，探討共培養及LED燈光照對牛樟芝(Antrodia cinnamomea)菌絲體生理、產物組成與產量之影響。前培養實驗確立牛樟芝最適生長環境，溫度25°C且環境酸鹼值於pH4和pH5時的生長直徑達到最佳的狀態。共培養三週後，牛樟芝分別與赤芝菌 (Ganoderma lucidum) 及蜜環菌 (Armillaria mellea) 之間會產生拮抗反應，在不同菌種之間會有深色代謝物的產生，相互作用下牛樟芝菌絲體有生理上的變化。不照光條件下，共培養之牛樟芝菌絲體生長會受到抑制，平均降低25-27%，藍光2.0W/m2照射後最高可增加14%。HPLC分析顯示牛樟芝經由LED紅光 (640-750nm) 照射有助於提高樟芝三萜類中去氫層孔菌酸(Dehydroeburicoic acid, Dea)及去氫硫色多孔菌酸(Dehydrosulphurenic acid, Dsa)的含量，牛樟芝與赤芝菌共培養會顯著增加樟芝酸B (Antcin B)、Dea及Dsa的含量，分別上升38%、35%及50%，於相同共培養條件下紅光照射會造成Dea及Dsa的含量降低。牛樟芝單培養條件下LED紅光照射有助於提高腺苷的含量，最高可達0. 5 mg/g，牛樟芝與赤芝菌共培養條件下牛樟芝的腺苷含量達到最高，由0.25 mg/g增加至0. 78 mg/g，共培養條件下照射紅光有礙於牛樟芝中腺苷的合成，藍光照射條件下皆會造成牛樟芝腺苷的含量降低。

This thesis is dedicated to the study of co-culture of Antrodia cinnamomea with two other fungal species, Ganoderma lucidum and Armillaria mellea, respectively, on the same solid culture environment. The optimal mycelial growth of A. cinnamomea were determined at 25oC and pH4-5. After about 3 weeks culture, a dark brownish strip made up by metabolite could be found between the two mutual antagonistic fungal colonies. The coculture systems were illuminated by red and blue LED light with dark as a control. The mycelial growth rate of A. cinnamomea was reduced 25-27% in dark, while blue light illumination enhances 14%. Red light illumination raised the levels of dehydroeburicoic acid (Dea) and dehydrosulphurenic acid (Dsa). Co-culturing with G. lucidum could increase the production of triterpenoids of A. cinnamomea, such as Actin B, Dea and Dsa for 38, 35 and 50%, respectively. However, the level of the terpenoids were reduced after red light illumination. Red light illumination may enhanced the production of adenosines of pure cultured A. cinnamomea to 0.5mg/g. Co-cultured with G. lucidum in dark improved the production of adenosine of A. cinnamomea from 0.25 mg/g to 0.78 mg/g. However, coculture with red and blue light illuminations inhibited the adenosine production of A. cinnamomea.

中文摘要 X
英文摘要 XI
第一章 前言 1
1.1 研究背景及動機 1
1.2 研究目的 3
第二章 文獻回顧 4
2.1 食用蕈類的介紹 4
2.2 蜜環菌背景介紹 4
2.3 赤芝背景介紹 5
2.4 牛樟芝背景簡介 7
2.4.1 牛樟芝-命名地位 7
2.4.2 牛樟芝-型態特徵 8
2.5 牛樟芝成分分析 12
2.5.1 三萜類化合物(Triterpenoides) 12
2.5.2 腺苷 (Adenosine) 19
2.5.3 多醣體 (Polysaccharides) 19
2.6 影響生長之環境因子 22
2.6.1 碳源及氮源 22
2.6.2 溫度 23
2.6.3 光照 23
2.6.4 pH值 25
2.6.5 競爭菌株共生培養 26
第三章 材料與方法 27
3.1 實驗架構 27
3.1.1 前實驗 27
3.1.2 主實驗-光照及共培養 28
3.2 材料 29
3.2.1 實驗菌株 29
3.2.2 藥品 30
3.2.3 標準品 31
3.2.4 儀器 31
3.2.5 光照相關設備 33
3.3 菌種培養 33
3.3.1 菌種保存 33
3.3.2 培養基製備 33
3.4 培養方法與條件 36
3.4.1 前培養 36
3.4.2 主培養 37
3.5 菌絲體物理評估 42
3.5.1 表面生長情形 42
3.6 化學成分分析 43
3.6.1 樣品收集及保存 43
3.6.2 三萜類 48
3.6.3 腺苷 47
3.6.4 總多醣 47
3.7 統計分析 50
第四章 結果與討論 51
4.1 前實驗-牛樟芝於不同條件下生長之情形 51
4.1.1 pH值對牛樟芝菌絲體生長之影響 51
4.1.2 溫度對牛樟芝菌絲體生長之影響 51
4.1.3 共生培養對牛樟芝菌絲體之影響 52
4.2 光照及共培養條件下生長之情形 55
4.2.1 不照光培養28天菌絲體生長之情形 55
4.2.2 紅光照射對菌絲體生長之影響 61
4.2.3 藍光照射對菌絲體生長之影響 74
4.3 三萜類成分分析 91
4.3.1 不照光條件下菌絲體三萜類成分分析 91
4.3.2 紅光照射條件下菌絲體三萜類成分分析 96
4.3.2 藍光照射條件下菌絲體三萜類成分分析 107
4.4 真菌菌絲體之腺苷含量 117
4.4.1 不同LED照射條件下菌絲體之腺苷含量 118
第五章 結論 122
參考文獻 124

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