臺灣博碩士論文加值系統

樟芝( Taiwanofungus camphoratus Wu et al.)是台灣特有的真菌，含有高產量之三萜類及其他生理活性物質，價格昂貴，目前難以人工栽培出子實體，是一種高價值的藥材。本研究發現樟芝雙核菌絲單核化的現象，挑選樟芝的雙核菌絲所產生的分節孢子(Arthroconidia)，進行培養後，利用玻片培養及cotton blue stain觀察，發現沒有扣子體( clamp-connection )的產生，推測為無性的單核孢子，利用此現象探討樟芝的交配型，分別挑選出節孢子進行培養，將單核無性孢子進行所有可能交配，結果所有的交配菌株皆沒有扣子體的產生。推測可能雙核菌絲形成節孢子時，只會釋出一種性別的核。利用單核孢子的交配於育種，分析無性單核菌絲所含三萜類成份，取含量高者進行交配，發現沒有加乘的作用。利用選種的方法分離出型態及分泌物不同的兩株菌株，分別為菌株7-1、7-4，其中菌株7-4具有一特有成分是菌株7-1所稀少或是缺乏的，透過TLC、PLC、HPLC、及液相層析質譜儀進行純化分析，推測此物質分子量為488。加入靈芝三萜類於培養基中，觀察樟芝菌絲體的形態並分析三萜類的生成，形態方面，加入少量的靈芝三萜類可以促進菌絲體孔洞的產生，透過光學顯微鏡及電子掃描顯微鏡觀察孔洞，並沒有發現有性世代的證據。三萜類分析方面，25℃培養180天仍然無法將靈芝三萜類轉換成樟芝三萜類。

Taiwanofungus camphoratus Wu et al. contained high triterpenoids and other physiologically active compounds were used as a precious herb and difficult to curture the fluting body with artificial environment in Taiwan. It was obvious that the clamp-connection was disappeared through the hyphae of the single-asexual-spore isolates and all the septate hypha containing only one nucleus with HCl-Giemsa stain. It suggested that the isolates were monokaryons. The mating reaction of isolates were now under investigation. It was obvious that no clamp-connection all the mating reaction. It was suggested dikaryon realese only one sexual nucleus to form asexual spore and then all the asexual spores had only one sexual spore. The mating reaction was not been observed. Even though, triterpenoid of all the monokaryon was analized by TLC, and triterpenoid of all the monokaryon was different. Triterpenoid contend of the after mating reaction with all monokaryon was not increased. Two different types 7-1 and 7-4 had the different appearance and TLC pattern were selected. A component found in mycelium 7-4 was purified with PLC and identified LC/MS/MS with molecular weight 488. T. camphoratus was cultured in PDA plate with additional triterpenoid of ganoderma tsuae in 25℃ for 180 days. It was obvious that pore formation were observed on the surface of mycelium. However, T. camphoratus mycelium could not transfer triterpenoid of Ganoderma tsugae.

中文摘要……..........................Ⅱ
英文摘要……………......................Ⅲ
致謝………………………………………………..
目錄………………………………………………..1
第一章 緒論………………………………………2
第一節、文獻整理……………………………..2
第二節、研究動機與目的……………………..23
第二章 實驗架構…………………………………24
第三章 研究材料與方法…………………………25
第一節、實驗材料與儀器……………………..25
第二節、實驗方法……………………………..27
第四章 結果………………………………………33
第五章 討論與結論………………………………36
第六章 參考文獻…………………………………40
圖…………………………………………………..44

第六章、參考文獻
1.Zang M, Su CH.: Ganoderma comphoratum, a new taxon in genus Ganoderma from Taiwan. China Acta Bot Yunnancia, 1990:395-396.
2.Chang TT, Chou WN: Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. Mycological Research 1995, 99(6):756-758.
3.Wu SH, Ryvarden L, Chang TT: Antrodia camphorata ('niu-chang-chih'), new combination of a medicinal fungus in Taiwan. Botanical Bulletin of Academia Sinica 1997, 38(4):273-275.
4.Wu SH, Yu ZH, Dai YC, Chen CT, Su CH, Chen LC, Hsu WC, Hwang GY: Taiwanofungus, a polypore new genus. Fung Sci 2004, 19:109-116.
5.張東柱: 牛樟之病害, 牛樟生物學及育林技術研討會論文集. 林業叢刊 1997, 72:127-131.
6.許勝傑, 陳清農,陳勁初: 樟芝宿主專一性之探討. 台灣農業化學與食品科學 2000, 38:533-539.
7.陳清農, 高炳煌,楊梅春,陳勁初: 台灣森林中的紅寶石樟芝. 鄉間小路 1999, 22.
8.王伯徹, 陳啟楨,華傑: 食藥用菇類的培養與應用. 食品工業發展研究所: 第M-87-019號 1998:187 頁.
9.Chang TT, Chou W.N.: Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. 1995, 99(Mycol. Res.):756-758.
10.廖英明: 菇類中的許不了~樟芝. 農業世界 1998, 176:76-79.
11.Lee IH, Chen CT, Chen HC, Hsu WC,Lu MK: Sugar flux in response to carbohydrate-feeding of cultured Antrodia camphorata, a recently described medicinal fungus in Taiwan. J of Chinese Medicine 2002, 13(1):21-31.
12.陳啟楨, 蘇慶華,藍明煌: 樟芝固體栽培其生物活性研究. Fung Sci 2001, 16(1,2):65-72.
13.陳勁初, 林文鑫,陳清農,許勝傑, 黃仕政,陳炎鍊,: 台灣特有真菌-樟芝菌絲體之開發. Fung Sci 2001, 16:7-22.
14.李學勇: 基礎植物學. 台北市: 國立編譯館; 1990.
15.易希珍: 普通植物學. 台北市: 環球出版社; 1976.
16.Cherng IH, Wu DP, Chiang HC: Triterpenoids from Antrodia cinnamomea. Phytochemistry 1996, 41(1):263-267.
17.Chang CY, Lee CL, Pan TM: Statistical optimization of medium components for the production of Antrodia cinnamomea AC0623 in submerged cultures. Appl Microbiol Biotechnol 2006.
18.Lu MK, Cheng JJ, Lai WL, Lin YR, Huang NK: Adenosine as an active component of Antrodia cinnamomea that prevents rat PC12 cells from serum deprivation-induced apoptosis through the activation of adenosine A(2A) receptors. Life Sci 2006.
19.Mizuno T, Hagiwara, T., Nakamura, T., Ito, H., Shimura, K., Sumiya, T. and Asakura, A.: Antitumor activity and some properties of water-soluble polysaccharides from “Himematsutake,” the fruiting body of Agaricus blazei murill. Agric Biol Chem 1990, 54:2889-2896.
20.Liu JJ, Huang, T. S., Hsu, M. L., Chen, C. C., Lin, W. S., Lu, F. J. and Chang, W. H.: Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. Toxicology & Applied Pharmacology 2004, 201:186-193.
21.李一宏: 樟芝菌絲體之培養及其多醣體抗乙型肝炎病毒活性評估. 中國醫藥學院中國藥學研究所; 2003.
22.黃鈴娟: 樟芝與姬松茸之抗氧化性質及其多醣組成分析. 國立中興大學 食品科學系; 2000.
23.范貞綺: 樟芝固態栽培與液態發酵菌絲體之成份及其活性之研究. 南台科技大學化學工程研究所; 2003.
24.蔡雁暉: 樟芝深層培養液及其多醣體之抗氧化特性. 國立中興大學食品科學研究所; 2002.
25.李一宏: 樟芝菌絲體之培養及其多醣體抗乙型肝炎病毒活性評估,. 2003.
26.Turner, W.B.and Aldridge, D.C.: Fungus Metabolies II; 1983.
27.Manitto: Biosynthesis of Natural products,. 1981:319.
28.高曉薇: 台灣靈芝屬新種樟芝之三萜類成分研究. 台北醫學院天然物
醫學研究所碩士論文; 1992.
29.Chen CH, Yang SW, Shen YC: New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum. Journal of Natural Products 1995, 58(11):1655-1661.
30.Yang SW, Shen YC, Chen CH: Steroids and triterpenoids of Antrodia cinnamomea - A fungus parasitic on Cinnamomum micranthum. Phytochemistry 1996, 41(5):1389-1392.
31.Cherng IH, Chiang HC, Cheng MC, Wang Y: Three new triterpenoids from Antrodia cinnamomea. Journal of Natural Products (Lloydia) 1995, 58(3):365-371.
32.Shen CC, Kuo YC, Huang RL, Lin LC, Don MJ, Chang T T , Chou CJ: New ergostane and lanostane from Antrodia camphorata. J Chin Med 2003, 14(4):247-258.
33.Shen YC, Wang YH, Chou YC, Chen CF, Lin LC, Chang TT, Tien JH, Chou CJ: Evaluation of the anti-inflammatory activity of zhankuic acids isolated from the fruiting bodies of Antrodia camphorata. Planta Medica 2004, 70(4):310-314.
34.Huang LC, Huang, S. J., Chen, C. C. and Mau, J. L.: Antioxidant properties of Antrodia camphorate, Proceedings of 3rd International Conference on Mushroom Biology & Mushroom Products. Proceedings of 3rd International Conference on Mushroom Biology & Mushroom Products 1999:275-283.
35.Hseu YC, Chang WC, Hseu YT, Lee CY, Yech YJ, Chen PC, Chen JY, Yang HL: Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sci 2002, 71(4):469-482.
36.Song TY, Yen GC: Antioxidant properties of Antrodia camphorata in submerged culture. J Agric Food Chem 2002, 50(11):3322-3327.
37.Hsiao G, Shen MY, Lin KH, Lan MH, Wu LY, Chou DS, Lin CH, Su CH, Sheu JR: Antioxidative and hepatoprotective effects of Antrodia camphorata extract. J Agric Food Chem 2003, 51(11):3302-3308.
38.魏佳瑜: 固體栽培樟芝甲醇萃取物抗變異自由基功效與
抗血小板凝集作用探討. 台北醫學大學公共衛生學研究所碩士論文; 2003.
39.簡淑娟: 人工固體栽培樟芝成分抽離暨自由基清除
抗變異活性探討. 臺北醫學大學公共衛生學系碩士班; 2005.
40.Nakamura N, Hirakawa A, Gao JJ, Kakuda H, Shiro M, Komatsu Y, Sheu CC, Hattori M: Five new maleic and succinic acid derivatives from the mycelium of Antrodia camphorata and their cytotoxic effects on LLC tumor cell line. J Nat Prod 2004, 67(1):46-48.
41.Liu JJ, Huang TS, Hsu ML, Chen CC, Lin WS, Lu FJ, Chang WH: Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. Toxicol Appl Pharmacol 2004, 201(2):186-193.
42.Peng CC, Chen KC, Peng RY, Su CH, Hsieh-Li HM: Human urinary bladder cancer T24 cells are susceptible to the Antrodia camphorata extracts. Cancer Lett 2006.
43.Hsu YL, Kuo YC, Kuo PL, Ng LT, Kuo YH, Lin CC: Apoptotic effects of extract from Antrodia camphorata fruiting bodies in human hepatocellular carcinoma cell lines. Cancer Lett 2005, 221(1):77-89.
44.Yang HL, Chen CS, Chang WH, Lu FJ, Lai YC, Chen CC, Hseu TH, Kuo CT, Hseu YC: Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by Antrodia camphorata. Cancer Lett 2006, 231(2):215-227.
45.陳勁初、陳建州、張基煌、毛正倫、喬長誠: CCRC36396 姬松茸與樟芝菌絲體之毒性與致突變性研究. 中華保健食品學會第二屆第一次會員大會 2001.
46.陳勁初、林文鑫、陳清農、許勝傑、黃仕政、陳炎鍊: 台灣特有真菌─樟芝菌絲體芝開發. Fung Sci 2001, 16(1,2):7-22.
47.林文川、郭淑卿、吳岳文: 樟芝菌株（CCRC 93032）發酵液對大鼠28天餵食的毒性作用. J Chin Med 2001, 12(4):293-303.
48.郭淑卿: 樟芝發酵液對大鼠肝臟纖維化及胃腸功能之改善作用. 中國醫藥學院中國藥學研究所藥學碩士論文，; 2002.
49.Su CH, Yang YZ, Ho HO, Hu CH, Sheu MT: High-performance liquid chromatographic analysis for the characterization of triterpenoids from Ganoderma. Journal of Chromatographic Science 2001, 39(3):93-100.
50.Shen YC, Wang YH, Chou YC, Chen CF, Lin LC, Chang TT, Tien JH, Chou CJ: Evaluation of the anti-inflammatory activity of zhankuic acids isolated from the fruiting bodies of Antrodia camphorata. Planta Med 2004, 70(4):310-314.