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研究生:張博仁
研究生(外文):Po-Jen Chang
論文名稱:仙草(MesonaprocumbensHemsel.)組織培養及其細胞萃取物抗氧化力之研究
論文名稱(外文):STUDIES ON TISSUE CULTURE AND THE ANTIOXIDATION PROPERTY OF CELL EXTRACT OF MESONA PROCUMBENS HEMSEL.
指導教授:何錦玟
指導教授(外文):Dr.Chin-Wen Ho
學位類別:碩士
校院名稱:大同大學
系所名稱:生物工程研究所
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:100
中文關鍵詞:仙草組織培養抗氧化
外文關鍵詞:Mesona procumbens Hemseltissue cultureantioxidation
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將桃園一號仙草頂芽及側芽置於含有不同濃度BA之MS基本鹽類培養基中,誘導不定芽產生,再將不定芽移至含有不同濃度GA3之培養基中,使不定芽抽長。結果在0.5 mg/l BA時會有最佳的不定芽誘導率(培養五週後,不定芽數目可增加31 ± 2.3倍),而在含有2 mg/l GA3之培養基中有最佳之不定芽抽長數(培養五週後,平均有76.6 ﹪之不定芽可抽長至3公分以上),而發根則是移至不含有植物生長調節劑之MS培養基中即可達到發根之效果,經過此過程培養植株可以順利出瓶健化,出瓶成功率可達80 ﹪,且植株並無變異產生,由不定芽誘導至出瓶健化所經歷時間僅需要12週。
而在經由不同培養基測試誘得之21個細胞系,萃取其total phenol,並比較清除自由基能力之測試,以培養於B5 + 1 mg/l Kinetin之培養基中誘導之癒合組織其清除能力可在系統濃度100μg/ml達到91.4 ﹪,且在系統濃度只有20μg/ml時,也可以達到64.8 ﹪之清除自由基能力。顯示此細胞萃取物具有高抗氧化能力。
The adventitious buds induced from shoots of Mesona procumbens Hemsel. cultured on MS basal medium supplemented with different plant growth regulators. The best proliferated ratio of adventitious buds was achieved 31±2.3 times when shoots were cultured on the medium contained with 0.5 mg/l BA. The maximum rate of elongation of adventitious buds was cultured on MS medium supplemented with 2 mg/l GA3 and the height of 76.6 % shoots was more than 3 cm. The root formation of adventitious buds more fast when transferred to hormone free medium. The survival percentage of plantlets was eighty percent and no visible variation after acclimation. The plantlets regeneration needs 12 weeks.
Twenty one cell lines were induced from different cultural media for selection of fast growing cells and the total phenol in cells was extracted to evaluate the scavenging free radical effect by DPPH test. The water extract from cells was cultured on B5 modified media with 1 mg/l kinetin expressed highly antioxidation property nearly 90 % under the system concentration was 100 μg/ml. It is showed the scavenging free radical effect up to 64.8 % even the system concentration was only 20μg/ml. These data indicated that cell extract from tissue culture of Mesona procumbens Hemsel. has highly antioxidation activity.
誌謝……………………………………………………………………….i
中文摘要…………………………………………………………………ii
ABSTRACT……………………………………………………………..iii
LIST OF TABLES……………………………………………………..viii
LIST OF FIGURES……………………………………………………...ix
ABBREVIATIONS……………………………………………………..xii
一、 前言……………………………………………………………..1
二、 前人研究………………………………………………………..4
2.1仙草名稱沿革……………………………………………………..4
2.2 栽培方式與品種選殖…………………………………………….4
2.3 唇形花科植物組織培養之相關研究……………………………5
2.3.1植株再生………………………………………………………6
2.3.1.1鼠尾草屬(Salvia)………………………………………6
2.3.1.2薄荷屬( Mentha )…………………………………………6
2.3.1.3薰衣草屬( Lavandula )…………………………………...9
2.3.2癒合組織誘導與二次代謝產物生成………………………..14
2.3.2.1鼠尾草屬(Salvia)…………………………………….14
2.3.2.2薄荷屬( Mentha )………………………………………..18
2.3.2.3薰衣草屬( Lavandula )………………………………….20
三、 材料與方法……………………………………………………22
3.1 植物材料………..……………………………………………….22
3.2無菌培養系統之建立……………………………………………22
3.2.1培植體種類與消毒方式……………………………………..22
3.2.2培養基………………………………………………………..22
3.2.3培養環境……………………………………………………..25
3.3癒合組織之誘導………………………………………………….25
3.3.1芽體培養……………………………………………………..25
3.3.2葉片培養……………………………………………………..25
3.3.3節間培養……………………………………………………..27
3.3.4光照強度對液體培養之癒合組織鮮重增加的影響………..27
3.4器官分化誘導.............................................................................…27
3.4.1生長調節劑對仙草癒合組織器官分化之影響……………..27
3.4.2植物生長調節劑對於仙草之不定芽增殖之影響…………..36
3.4.3 GA3對於仙草不定芽抽長之影響…………………………..36
3.4.4 NAA對仙草不定芽之發根影響……………………………38
3.5再生植株之出瓶與健化………………………………………….38
3.6仙草癒合組織增殖與其水萃取物清除自由基能力之相關性….41
3.6.1癒合組織之增殖比較………………………………………..41
3.6.2癒合組織水萃取物之清除自由基能力測定………………..41
3.6.2.1癒合組織之成份萃取……………………………….…..41
3.6.2.2清除DPPH自由基能力之測定…………………….…..45
3.7形態觀察與照相………………………………………………….45
3.8統計分析………………………………………………………….45
四、 結果……………………………………………………………46
4.1癒合組織之誘導……………………………………………….....46
4.1.1芽體培養……………………………………………………..46
4.1.2葉片培養……..………………………………………………46
4.1.3節間培養………………………………………………….….46
4.1.4光照強度對液體培養之癒合組織鮮重增加的影響………..50
4.2器官分化誘導.................................................................................50
4.2.1生長調節劑對仙草癒合組織器官分化之影響.…………….50
4.2.2植物生長調節劑對於仙草之不定芽增殖之影響.………….54
4.2.3 GA3對於仙草不定芽抽長之影響…..………………………59
4.2.4 NAA對仙草不定芽之發根影響………….………………...59
4.3再生植株之出瓶與健化……………………………….…………64
4.4仙草癒合組織增殖與水萃取物清除自由基能力之相關性…….64
4.4.1癒合組織之增殖比較………………………………………..64
4.4.2癒合組織水萃取物之清除自由基能力……….…………….70
4.4.2.1癒合組織之成份萃取…………………………………...70
4.4.2.2清除DPPH自由基能力之測定……………………...…72
五、 討論……………………………………………………………79
5.1癒合組織之誘導…………………………………………….……79
5.1.1不同培植體對癒合組織誘導之影響………………………..79
5.1.2光照強度對液體培養之癒合組織鮮重增加的影響………..80
5.2器官分化誘導.................................................................................80
5.2.1生長調節劑對仙草癒合組織器官分化之影響.…………….80
5.2.2植物生長調節劑對於仙草之不定芽增殖之影響.…..……...81
5.2.3 GA3對於仙草不定芽抽長之影響…………………………..82
5.2.4 NAA對仙草不定芽之發根影響………………………....…82
5.3再生植株之出瓶與健化………………………………..………...83
5.4仙草癒合組織增殖與其水萃取物清除自由基能力相關性…….83
六、 結論……………………………………………………………85
參考文獻……………………………………………………………….86
1. 甘偉松,1980. 藥用植物等. 國立中國醫藥研究所. 台北.
p.482
2. 李勉民,1998. 常用藥草圖說. 讀者文摘. 香港. p.283
3. 邱年永、張永光,1986. 原色台灣藥用植物圖鑑(2). 南天書局.台北. p.190
4. 邱年永,1991. 百草茶原植物. 宏祥出版社. 台中. p.90
5. 林俊彥,1999. 台灣保健植物栽培及利用. 桃園農業改良場. 桃
園. p.52-53
6. 姜金龍、史宏財、龔財立,1991. 植苗數與氮肥施量對仙草產量與品質之影響. 桃園區農業改良場研究報告. 8:1-8
7. 姜金龍,1995. 仙草. 增修訂再版台灣農家要覽農作篇(一). 農
業委員會台灣農家要覽增修訂再版策劃委員會. 豐年社. 台北. p.183-186
8. 姜金龍、龔財立、辛仲文,1997. 栽培密度對仙草產量與品質之影響. 桃園區農業改良場研究報告. 31:13-17
9. 姜金龍、龔財立、辛仲文,1999. 桃園區少量多樣化農特產專輯.
桃園區農業改良場發行. p.71-73
10. 姜金龍,1999. 台灣野生仙草種內變異之研究. 博士論文. 國立中興大學農藝學系.
11. 姜金龍、龔財立、辛仲文、林俊清,2000. 仙草「桃園一號」之育成. 桃園區農業改良場研究報告. 42:1-14
12. 胡敏夫、林禮輝,1986. 仙草不同生長期之主要成份含量分析. 中華農業研究. 35(2):180-185
13. 胡敏夫,1988. 仙草. 經濟植物集. 梁鶚(林果)主篇. 豐年社.
台北. p.160-163
14. 胡敏夫、姜金龍、龔財立、呂秀英、劉新裕,1998. 仙草優良品系區域適應性試驗. 中華農業研究. 47(1):1-11
15. 胡敏夫、劉新裕、羅淑卿、盧煌勝, 2000. 仙草新品種農試一號之育成. 中華農業研究. 49(1):12-25
16. 洪千雅,2001. 仙草抗氧化機能性之研究. 博士論文. 國立中興
大學食品科學研究所.
17. 許喬木、邱年永,1989. 原色野生食用植物圖鑑. 南天書局. 台
北. p.185
18. 陳勝彰、曾富生,1990a. 台灣旱田雜草香附子種內變異研究. I.
田間與田埂族群之形態特性變異. 農林學報. 39(1):103-116
19. 陳勝彰、曾富生,1990b. 台灣旱田雜草香附子種內變異研究.Ⅱ. 七個地區族群之生態特性之變異. 中華民國雜草學會會刊. 11: 45-61
20. 楊在義,1982. 台灣植物名彙. 天然書社. 台北. p.1121
21. 楊恭毅,1984. 楊氏園藝植物大名典. 中國花卉雜誌社. 台北.
p.4922-4923
22. 蔡振聰、呂秋菊、鄭元春、邵亨言,1985. 台灣特用植物圖鑑.
台灣省立博物館. 台北. p.190
23. Andrade L.B., S. Echeverrigaray, F. Fracaro, G.F. Pauletti and L. Rota. 1999. The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC.). Plant cell, tissue and organ culture.56:79-83
24. Anna M.S., M.Panizza and F.Tognoni. 1995. Endogenous ethylene requirement for adventitious root induction and growth in tomato cotyledons and lavandin microcuttings in vitro. Plant growth regulation. 17:205-212
25. Bolta Z., D. Baricevic, B. Bohanec and S.Andrensek. 2000. A preliminary investigation of ursolic acid in cell suspension culture of Salvia officinalis . Plant cell, tissue and organ culture. 62: 57-63
26. Caissard J.C., O.Faure, F. Jullien, M. Colson. And A. Perrin. 1996. Direct regeneration in vitro and transient GUS expression in Mentha ×piperita. Plant cell reports. 16:67-70
27. Calvo M.C. and J. Segura. 1988. In vitro morphogenesis from explants of Lavandula latifolia and Lavandula stoechas seedings. Scientia horticulturae. 36:131-137
28. Calvo M.C. and J. Segura. 1989a. Plant regeneration from cultured leaves of Lavandula latifolia medicus :Influence of growth regulators and illimination conditions. Plant cell ,tissue and organ culture. 19:33-42
29. Calvo M.C. and J. Segura. 1989b. In vitro propagation of lavender. Hortscience. 24:375-376
30. Chang J.M., J.H. Shin, I.N. Chung and H.J. Lee. 1998.Improved menthol production from chitosan-elicited suspension culture of Mentha piperita. Biotechnology letter. 20(12):1097-1099
31. Chen C.W. and C.T. Ho. 1997. Antioxidant activities of caffeic acid and its related hydroxycinnamic acid compounds. Journal of agricultural and food chemistry. 45:2374-2378
32. Chen H. and F. Chen. 1999a. Effects of methyl jasmonate and salicylic acid on cell growth and cryptotanshinone formation in Ti transformed Salvia miltiorrhiza cell suspension cultures. Biotechnology letter. 21: 803-807
33. Chen H. and F. Chen. 1999b. Kinetics of cell growth and secondary metabolism of a high-tanshinone-producing line of the Ti transformed Salvia militiorrhiza cell in suspension cultures. Biotechnology letters.21:701-705
34. Chen H. and F. Chen. 2000a. Effect of yeast elicitor on the secondary metabolism of Ti-transformed Salvia militiorrhiza cell suspension cultures. Plant cell reports.19:710-717
35. Chen H. and F. Chen. 2000b. Effect of yeast elicitor on the growth and secondary metabolism of a high — tanshinone - producing line of the Ti-transformed Salvia militiorrhiza cell in suspension cultures. Process biochemistry. 35:837-840
36. Chen H., F. Chen, Y.L. Zhang and J.Y. Song. 1999a. Production of rosmarinic acid and lithospermic acid B in Ti transformed Salvia militiorrhiza cell suspension cultures. Process biochemistry. 34:777-784
37. Chen H., F. Chen, Y.L. Zhang and J.Y. Song. 1999b. Production of lithospermic acid B and rosmarinic acid in hairy root cultures of Salvia militiorrhiza. Journal of industrial microbiology and biotechnology. 22:133-138
38. Cheng W.T. 1990. Population studies on Formosan Lilium ( Liliaceae) Ⅰ.A cluster analysis of variants in L. longiflorum Thunb. Taiwania. 35:198-205.
39. Chou C.H., S.Y. Hwang and F.C. Chang.1987. Population study of Miscanthus floridulus (Labill.) Warb. I.Variation of peroxidase and esterase in 27 populations in Taiwan. Botanical bulletin of academia sinica. 28:247-281.
40. Chou C.H. and F.C. Chang. 1988. Population study of
Miscanthus floridulusⅡ. Ecotypic variation of M. floridulus and M. transmorri-sonensis as affected by altitude in Nantou, Taiwan. Botanical bulletin of academia sinica. 29:301-314.
41. Chou C.H. and Y.Y. Chen. 1990. Population study of Miscanthus floridulus ⅢPopulation variation of M. floridulus in Green and Orchid islets of Taiwan . Botanical bulletin of academia sinica. 31:223-233.
42. Cuvelier M.E., H. Richard and C. Berset. 1996. Antioxidative activity and phenolic composition of pilot-plant and commercial extracts of sage and rosemary. JAOCS. 73:645-652
43. Dronne S., M. Colson, S. Mpja and O. Faure. 1999. Plant regeneration and transient GUS expression in a range of lavandin (Lavandula × intermedia Emeric ex Loiseleur) cultivars. Plant cell ,tissue and organ culture. 55:193-198
44. Duncan D.B. 1955. Multiple range and multiple F-test. Biometrics. 11:1-42
45. Faure O., F. Diemer, S. Moja and F. Jullien. 1998. Mannitol and thidiazuron improve in vitro shoot regeneration from spearmint and peppermint leaf disks. Plant cell,tissue and organ culture. 52:209-212
46. Gamborg O.L.,R.A.Miller and K.Ojima. 1968. Nutrient requirements of suspension culture of soybean root Cells.Exp. Cell Res.50:151-158
47. Hilton M.G., A. Jay, M.J.C. Rhodes and P.D.G. Wilson. 1995. Growth and monoterpene production by transformed shoot cultures of Mentha citrata and Mentha piperita in flasks and fermenters. Appl microbiol bioltechnol. 43:452-459
48. Hippolyte I., B. Marin, J.C. Baccou and R. Jonard. 1992. Growth and rosmarinic acid production in cell suspension cultures of Salvia officinalis L. Plant cell reports. 11:109-112
49. Hohmann J., I. Zupko, D. Redei, M. Csanyi, G. Falkay, I. Mathe and G. Janicsak. 1999. Protective effects of the aerial parts of salvia officinalis, Melissa officinalis and Lavandula angustifolia and their constituents against enzyme-dependent and enzyme-independent lipid peroxidation. Planta medica. 65:576-578
50. Hosoki T. and Y. Tahara.1993. In vitro propagation of Salvia leucantha Cav. HortScience. 28(3):226
51. Huang M.T.,C.T. Ho, Z.Y. Wang, T. Ferraro, Y.R. Lou, K. Stauber, W. Ma, C. Georgiadis, J.D. Laskin and A.H. Conney. 1994. Inhibition of skin tumorigenesis by rosemary and its constituents carnosol and ursolic acid. Cancer research. 54:701-708
52. Jie L. 1995. Pharmacology of oleanolic acid and ursolic acid. J. Ethnopharmacol. 49:57-68
53. Jullien F., F. Diemer, M. Colson and O. Faure. 1998. An optimizing protoplast regeneration of three peppermint cultivars ( Mentha × piperita). Plant cell,tissue and organ culture. 54:153-159
54. Kashiwada Y., H.K. Wang, T. Nagao, S. Kitanaka, I. Yasuda, T. Fujioka, T. Yamagishi, M.L. Cosentino, M. Kozuka, H. Okabe, Y. Ikeshiro, C.Q. Hu, E. Yeh and K.H. Lee. 1998. Anti-HIV activity of oleanolic acid , pomolic acid and structuraly related triterpenoids. Journal of natural products. 61:1090-1095
55. Kazuki H., K. Shinichi and Y. Michio. 1996. Natural antioxidants and active oxygen removers in food cosmetics, and pharmaceuticals. Chemical abstracts. 124:200690r
56. Kintzios S., A. Nikolaou and M. Skoula. 1999. Somatic embryogenesis and in vitro rosmarinic acid accumulation in Salvia officinalis and S. fruticosa leaf callus cultures. Plant cell report. 18:462-466
57. Li X.,X. Niu, R.A. Bressan, S.C. Weller and P.M. Hasegawa. 1999. Efficient plant regeneration of native spearmint ( Mentha spicata L.). In vitro cellular & developmental biology : journal of the tissue culture association 35:333-338
58. Liu J. 1995. Pharmacology of olanolic acid and ursolic acid . J. Ethnopharmacol. 49:57-68
59. Masaki H.,S. Sakaki, T. Atsumi and H. Sakurai. 1995. Active-oxygen scavenging activity of plant extracts. Biol. Pharm. Bull. 18(1):162-166
60. Miguel C.S.G. and M.C. Calvo. 1996. Micropropagation of Lavandula latifolia through nodal bud culture of mature plants. Plant cell ,tissue and organ culture. 45:259-261
61. Moon J. H. and J. Terao. 1998. Antioxidant activity of caffeic acid and dihydrocaffeic acid in lard and human low-density lipoprotein. Journal of agricultural and food chemistry. 46: 5062-5065.
62. Murashige T.and F.Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant. 15:473-497
63. Niu X., K. Lin, P.M. Hasegawa and R.A. Bressan. 1998. Transgenic peppermint(Mentha × piperita L.)plants obtained by cocultivation with Agrobacterium tumefaciens. Plant cell reports. 17:165-171
64. Panizza M. and F. Tognoni. 1988. Clonal propagation, callus
formation and plant regeneration of lavandin. Scientia horticulturae. 37:157-163
65. Paula C., S. Gomes, R.M. Seabra, P.B. Andrade,and M.F. Ferreira. 2002. Phenolic antioxidant compounds produced by in vitro shoots of sage( Salvia officinalis L. ). Plant science.162: 981-987
66. Pavlov A. and M. Ileva. 1999.The influence of phenylalanine on accumulation of rosmarinic and caffeic acids by Lavandula vera MM cell culture. World journal of microbiology & biotechnology. 15:397-399
67. Pavlov A.I., M.P. Ileva and I.N. Panchev. 2000.Nutrient medium optimization for rosmarinic acid production by Lavandula vera MM cell suspension. Bioltechol. Prog. 16:668-670
68. Phatak S.V. and M.R. Heble. 2002. Organ genesis and terpenoid synthesis in Mentha arvensis. Fitoterapia. 73:32-39
69. Quazi M.H. 1980. In vitro multiplication of Lavandula spp. Annals of botany. 45:361-362
70. Recio C.M., R.M. Giner, S. Manez, J.Gueho, H.R. Julien, K. Hostettmann and J.L. Rios. 1995. Investigation on the steroidal anti-inflammatory activity of triterpenoids from Diospyros leucomelas. Planta medica. 61:279-285
71. Sato H., E. Sueo , O. Seibi , H. Kazuo and I. Yoshio. 1993. Plant regeneration from protoplasts of peppermint ( Mentha pipperita L.). Plant cell reports. 12:546-550
72. Sidria C., M.T. Pinol, J. Palazon, R.M. Cusido, R. Vila, C. Morales, M. Bonfill and S. Canigueral. 1999. Influence of plant growth regulators on the growth and essential oil content Lavandula dentate plantlets. Plant cell, tissue and organ culture. 58:177-184
73. Suga T., T. Hirata and Y. Yamamoto. 1980. Liqid constituents of callus of Mentha spicata. Agricultural and biological chemistry. 44:1817~1820
74. Tal. B., J.S. Rokem and I. Goldberg. 1983. Factors affecting growth and product formation in plant cells growth in continuous culture. Plant cell reports. 2:219-222
75. Ternes W. and K. Schwarz. 1995. Antioxidat constituents of Rosmarinus officinalis and Salvia officinalis Ⅳ. determination of carmosic acid in different foodstuffs. Z. Lebensm. Unters. Forsch. 201:548-550
76. Tisserat B. and R. Silman. 2000. Interactions of culture vessels, media volume, culture density, and carbon dioxide levels on lettuce and spearmint shoot growth in vitro. Plant cell reports. 19:464-471
77. Tsuro M., M. Koda and M. Inoue. 1999. Comparative effect of different types of cytokinins for shoot formation and plant regeneration in leaf-derived callus of lavender (Lavandula vera DC.). Scientia horticulturae. 81:331-336
78. Tsuro M., M. Koda and M. Inoue. 2000. Efficient plant regeneration from multiple shoots formed in the leaf-derived callus of Lavandula vera, using the “open culture system”. Scientia horticulturae. 86:81-88
79. Tsuro M., M. Inoue and H. Kameoka. 2001.Variation in essential oil components in regenerated lavender (Lavandula vera DC) plants. Scientia horticulturae. 88:309-317
80. Van Eck J.M. and S.L. Kitto. 1992. Regeneration of peppermint and orange mint from leaf disks. Plant cell,tissue and organ culture. 30:41-49
81. Watanabe K., H. Mitsuda and Y. Yamada. 1983. Retention of metabolic and differentiation potential of green Lavandula vera callus after Freeze-Preservation. Plant & cell physiol. 24(1):119-122
82. Yen G.C. and J.C. Wu. 1999. Antioxidant and radical scavenging properties of extracts from Ganoderma tsugae. Food chemistry. 65:375-379
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