玄參自古即為我國常用中藥之一，最早收載於神農本草經中品，其主要的活性成分玄參皂苷(harpagoside)，對於抗菌、咽喉炎、便秘具有顯著的療效。本試驗主要目的是將取得的三種主要作為玄參藥材的植物，包括北玄參、浙玄參和臺灣產之雙鋸齒葉玄參，進行組織培養之大量繁殖、癒合組織培養，以及成分測定的研究，藉以探討三種玄參之組織培養和玄參皂苷累積的情形，結果簡述如下：
1. 將北玄參之莖節及浙玄參和雙鋸齒葉玄參之頂芽，分別培養於添加1.0 mg/l、0.5~1.0 mg/l、1.0~4.0 mg/l BA之MS固體培養基中，可獲得大量增殖之芽體。
2. 將北玄參和浙玄參之芽體，接種於以16層藥包紙覆蓋瓶口的MS固體培養基中，其組織培養苗存活率最高；雙鋸齒葉玄參之芽體，以接種於覆蓋8層藥包紙的處理最佳。
3. 將北玄參、浙玄參和雙鋸齒葉玄參葉片誘得癒合組織，分別繼代培養於含有0.5~8.0 mg/l、1.0~4.0 mg/l及1.0 mg/l 2,4-D的培養基最佳，但所獲得的癒合組織皆無玄參皂苷的累積。
4. 分析北玄參、浙玄參和雙鋸齒葉玄參1~3個月的組織培養苗，以雙鋸齒葉玄參2個月的地上部玄參皂苷最高。

Xuan-Shan is an important medicinal plant of the family Scrophulariaceae and was first described in the Shen-Nong-Pen-Ts’ao-Chin under the middle category. The most important constituents of Xuan-Shan is harpagoside, which been used in the traditional Chinese medicine in the treatment of inflammation, laryngitis, tonsillitis, abscesses of carbuncles and constipation. S. ningpoensis is the most usual plant resource for the crude drug of Xuan-Shan in China. In Korea and Japan, S. buergeriana is resource for the crude drug of Xuan-Shan. S. yoshimurae is indigenous plant to Taiwan, a substitute for S. ningpoensis. The main objectives of present investigation were (1) to standardize a protocol for rapid multiplication of S. buergeriana, S. ningpoensis and S. yoshimurae from shoot-tip and nodal explants. (2) to standardize a protocol for establishment of callus culture using the leaf explants. (3) to analyze and compare the amount of harpagoside present in the callus, 3 months old in vitro cultured plantlet, marketed crude drug and marketed processed drug (‘KO DA’ Shyuan Shen, extract subtly granule).
The results obtained are summarizes as follow:
1. A simple and rapid complete plant regeneration protocol using shoot-tip and nodal explants of S. buergeriana, S. ningpoensis and S. yoshimurae has been standardized. The shoot organogenesis response varied, with the two explants tested in the various BA containing medium in all the three species. The best multiplication rate for S. buergeriana, 5.2 shoots per responding explants, was observed when the nodal segments were cultured on MS basal supplemented with 1mg/l BA. While in S. ningpoensis and S. yoshimurae maximum number of shoots developed (13.7 and 4.4 shoots per responding explant respectively) when the shoot tip explants were cultured on MS basal medium supplemented with BA respectively.
2. Hyperhydricity in the shoot cultures was reduced by using dispense paper as culture vessel closures. Healthy shoot growth could be achieved by covering the culture vessel with 16 layers of dispense paper for S. buergeriana and S. ningpoensis and 8 layers of dispense paper S. yoshimurae shoot cultures.
3. Callus was induced by culturing the leaf explants on MS nasal medium supplemented with various concentrations (0.5 — 16.0 mg/l) of 2, 4, D. The callus cultures of S. buergeriana, S. ningpoensis and S. yoshimurae are been successfully maintained by sub-culturing 200 mg of calli on fresh media containing 1.0 mg/l 2, 4 D, every month.
4. The high performance liquid chromatography (HPLC) analysis for the harpagoside contents in the callus, aerial parts of 2 months old in vitro grown plantlet of the three, roots of 2 month old in vitro grown plantlet of the three species, marketed crude drug, marketed processed drug revealed that the major medicinal compound -harpagoside. Content of harpagoside in aerial parts obtained from in vitro propagated plants in two months of Scrophularia yoshimurae is highest.

縮寫 1
前言 2
前人研究 4
材料與方法 21
結果 29
討論 55
中文摘要 59
英文摘要 60
參考文獻 62
附錄 75

參考文獻
小野蘭山。1928。重訂本草綱目啟蒙，第147-148頁。日本古典全集刊行會。
田蔚城。1996。生物技術（初版），第207-221頁。九州圖書文物有限公司。臺北市。
李承榆。2000。值得產業發展的農業生物技術。中華農藝學會簡訊。11：9。
李醫明，蔣山好，朱大元。1999。玄參屬植物化學成分與藥理活性研究進展。中草藥 30：307-310。
邱德文，吳家榮，夏同珩。2001。玄參。本草綱目彩色藥圖，第122頁。薪傳出版社，臺北市。
周立剛、鄭光植、王世林。1990。寡糖素對滇紫草癒傷組織色素合成的影響。天然產物研究與開發 2：22-25。
林弘敏，陳世銘，陳忠川，蔡新聲。1998。利用組織培養大量繁殖雙鋸齒葉玄參。中華農學會報 183：1-12。
林李昌。2003。何首烏之組織培養。國立中興大學農藝研究所碩士論文，臺中市。
林郁進。1995。黃耆類藥材之生藥及膜莢黃耆組織培養之研究。博士論文。臺中：中國醫藥學院中國藥學研究所。
南京中國醫藥大學。1999。玄參。中華本草（第七冊），第391-396頁。上海科學技術出版社，上海市。
高景輝。1998。植物荷爾蒙生理。臺北：華香園出版社。pp.25-43
徐剛標、何方、陳良昌。1999。銀杏癒合組織誘導與繼代培養的研究。中南林學院學報 19：32-36。
張東迪。1988。地黃組織培養之研究。碩士論文。臺中：中國醫藥學院中國藥學研究所。
張建春、朱建美。2003。玄參的化學成分與藥理活性研究進展。山東醫藥工業 22：25-27。
陳威臣、夏奇鈮、葉茂生、蔡新聲。2004。植物組織培養苗玻璃質化現象與低木質化作用關聯性之探討。科學農業 52(3,4)：90-96。
陳世銘。1996。雙鋸齒葉玄參組織培養之研究。中國醫藥大學中國藥學研究所碩士論文，臺中市。
陳益明。1988。植物荷爾蒙-生長素與激勃素。出自植物生長調節劑在園藝作物之應用研討會專輯，pp.15-41。農業試驗所。臺中。
陳和榮、陳敏、陳褔太、鐘風林。1986。藥用青蒿組織培養II。中藥通報 11：9-11。
葉豐次。1995。柴胡類之生藥學及竹葉紫胡組織培養之研究。博士論文。臺中：中國醫藥學院中國藥學研究所。
葉茂生、前川文夫、湯淺浩史。1983 マソ科イニケン族植物の染色體數。（財）進化生物學研究報告 2：37-44。
蔡新聲。1982。組織培養技術之研究及應用。中華農學會報 120: 9-21
臺灣大學植物系。1998。雙鋸齒葉玄參。臺灣植物誌（第二版），第624-625頁。國立臺灣大學植物系出版，臺北市。
劉新裕、林俊義、張城國。2002。藥用植物專輯。臺中:行政院農業委員會農業試驗所。pp.206-210
蕭培根、連文琰。1998。北玄參、玄參。原色中藥原植物圖鑑（下），第432-433。南天書局，臺北市。
Sagare, A. P., 李雅琳、蕭翌柱、陳威臣、蔡新聲。2000。臺灣珍稀藥用植物利用組織培養之繁殖技術。出自2000年海峽兩岸生物多樣性與保育研討會論文集，周延鑫、謝豐國、吳聲華、周文豪主編，pp:441-458。臺中：國立自然科博學物館。
Aitken, C. J., A. P. Singh, K. J. Horgan and T. A. Thorpe. 1985. Explant developmental state and shoot formation in Pinus radiata cotyledons. Bot. Gaz. 146:196-203.
Amano, A., K. Fujimoto, H. Ohashi, K. Matsunaga and H. Mizukami. 1989. Chromosome number variation in Bupleurum falcatum plants regenerated through somatic embryogenesis of callus cultures. Japan. J. Pharmac. 43:13-18.
Andrijany, V. S., G. Indrayanto and L. A. Soehono. 1999. Simultaneous effect of calcium, magnesium, copper and cobalt ions on sapogenin steroids content in callus cultures of Agave amaniensis. Plant Cell Tiss. Org. Cult. 55:103-108.
Arnold, S. V. and T. Eriksson. 1984. Effects of agar concentration on growth and anatomy of adventitious of Picea abies L. Plant Cell Tiss. Org. Cult. 3:257-264.
Arya, S., J. R. Liu and T. Eriksson. l991. Plant regeneration from protoplasts of Panax ginseng (C.A. Meyer) through somatic embryogenesis. Plant Cell Rep. 10:277-281.
Benjamin, B. D., P. C. Roja, M. R. Heble and M. S. Chadha. 1987. Multiple shoot cultures of Rauvolfia serpentina: growth and alkaloid production. J. Nat. Prod. 129:129—135.
Bourgaud, F., A. Gravot, S. Milesi and E. Gontier. 2001. Production of plant secondary metabolites: a historical perspective. Plant Sci. 5:839-851.
Cacho, M., M. Moran, M. T. Herrera, J. Fernandez-Tarrago and M. P. Corchete. 1991. Morphogenesis in leaf, hypocotyls and explants of Digitalis thapsi L. cultured in vitro. Plant Cell Tiss. Org. Cult. 25:117-123.
Chand, S., A. K. Sahrawat and D. V. S. S. R. Prakash. 1997. In vitro culture of Pimpinella anisum L. (anise). J. Plant Bioch. Biotech. 6:1—5.
Chee, R. and R. Pool. 1989. Effects of light quality on tissue culture. J. Am. Soci. Hort. Sci. 6:114-350.
Chen, L. J., T. W. Hu and L. C. Huang. 1995. A protocol toward multiplication of the medicinal tree, Eucommia ulmoides Oliver. In Vitro Cell. Dev. Biol.-Plant 31:193—198.
Christen, P., M. F. Roberts, J. D. Phillipson and W. C. Evans. 1989. High yield production of tropane alkaloids by hairy-root cultures of a Datura candida hybrid. Plant Cell Rep. 8:75-77.
Chueh, F. S., C. C. Chen and H. S. Tsay. 2000. Studies on factors affecting the establishment of Gentiana davidii var. formosana (Hayata) T. N. Ho. cell suspension cultures. J. Food & Drug Analysis 8:297-303.
Damgaard, O. and O. Rasmussen. 1991. Direct regeneration of transformed shoots in Brassica napus from hypocotyl infections with Agrobacterium rhizogenes. Plant Mol. Biol. 17:1-8.
De-Eknamkul, W. and B. E. Ellis. 1985. Effects of macronutrients on growth and rosmarinic acid formation in cell suspension cultures of Anchusa officinalis. Plant Cell Rep. 4:46-49.
Dornenburg, H. and D. Knorr. 1995. Strategies for the improvement of secondary metabolite production in plant cell cultures. Enzyme Microb. Technol. 17:674-684
Economou, A. S. 1987. Light treatment to improve efficiency of in vitro propagation systems. J. Japan. Soc. Hort. Sci. 22:751-754.
Erdei, I., Z. Kiss and P. Maliga. 1981. Rapid clonal multiplication of Digitalis lanata in tissue culture. Plant Cell Rep. 1:34-35.
Evans, D. A., W. R. Sharp and C. E. Flick. 1981. Growth and behavior of cell cultures : embryogenesis and organogenesis. In : Plant Tissue Culture: Methods and Applications in Agriculture. ed. T.A. Thorpe, pp. 45-113, New York, Academic Press.
Faria, R. T., and R. D. Illg. 1995. Micropropagation of Zingiber spectabile Griff. Sci. Hort. 62:135—137.
Fujioka, N., Y. Kurish, H. Miyagawa, H. Kohda, K. Yamasaki, R. Shoyama and I. Nishioka. 1986. Studies on the tissue culture of Panax japonicus (1) Multiplication by somatic embryogenesis of flower bud and rhizome. Japan. J. Pharmac.40:152-158.
Fujioka, N., K. Nahao, Y. Fujita, H. Miyagawa, H. Kohda, K. Yamasaki and S. Takami. 1983. The production of plant regenerated from shoot tips of Astragalus membranaceus. Japan. J. Pharmac. 37:405-411.
Fujita, Y., Y. Hara, C. Suga and T. Morimoto. 1981. Production of shikonin derivatives by cell suspension cultures of Lithospermum erthrorhizon. Plant Cell Rep. 1:61-63.
Fukui, H., M. Tani and M. Tabata. 1990. Induction of shikonin biosynthesis by endogenous polysaccharides in Lithospermum erythrorhizon cell suspension cultures. Plant Cell Rep. 9:73-76.
Gamborg, O. L., R. A. Miller and K. Ojima. 1968. Nutrient requirements of suspension cultures of soybean root cell. Exp. Cell. Rep. 50:151-158.
Gamborg, O. L., T. Murashige, T. A. Thorpe and I. K. Vasil. 1976. Plant tissue culture media. In Vitro 12:473—478.
Gandaeidaja, D. 1980. Effect of nitrate and ammonium on the growth of tissue culture of Dendrobium phalaeopsis Fitzg. Ann. Bot. 7:63-69.
George, E. F. and P. D. Sherrington. 1984. Plant Propagation by Tissue Culture. Handbook and Directory of Commercial Laboratories. Ed. England: Eastern Press, Reading, Berks. pp.125-330.
Giri, A. and M. L. Narasu. 2000. Transgenic hairy roots: recent trends and applications. Biotechnol. Adv. 18:1-22.
Handique, P.J. and P. Bora. 1999. In vitro regeneration of a medicinal plant- Houttuynia cordata Thunb. from nodal explants. Curr. Sci. 76:1245—1247.
Heberle-Bors, E. 1980. Interaction of activated charcoal and iron chelates in anther cultures of Nicotiana (tobacco) and Atropa belladonna. Z. Pflanzenphysiol. 99:339-347.
Hiraoka, H., N. Yamada, M. Oyanagi and Y. Tomitam. 1987. Vegetative propagation of Saposhnikoria divaricata through embryogensis of leaf callus. Japan. J. Pharmac. 41:43-47.
Hu, Z. B., and A.W.Alfermann.1993. Diterpenoid production in hairy root cultures of Salvia miltiorrhiza. Phytochemistry 32:699-703
Hu,C.Y., and P.J. M. Wang. 1983. Shoot-tip and bud culture. In: Plant Cell Culture, eds. D. A. Evans, W.R. Wang, P.V.Ammirato, and Y.Yamada, pp. 177—277. New York, MacMillan.
Huang, L. and T. Murashige. 1977. Plant tissue culture media: major constituents; their preparation and some applications. Tissue Culture Assoc Manual 13:539—548.
Irintoto, B., K. H. Tan and H. E. Sommer. 1993. Effect of humic acid on callus culture of slash pine (Pinus elliottii Engelm). J. Plant Nutr. 16:1109-1118.
Kamada, H., N. Oakmara, M. S. Harada and K. Shimomura. 1986. Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Rep. 5:239-242.
Kanrtnig, T. H., U. Russheim and B. Maunz. 1976. Cardenolide in Oberflachen kulturenaus and Keimund laubblattern von Digitalis purpurea. Planta Med. 29:275-282.
Kassanis, B. 1957. The use of tissue cultures to produce virus-free clones from infected potato varieties. Ann. Appl. Biol. 45:422-427.
Kitamura, Y., H. Miura and M. Sugii. 1989. Plant regeneration from callus culture of Swertia pseudochinensis. Japan. J. Pharmac. 43:256-258.
Komalavalli, N. and M. V. Rao. 2000. In vitro micropropagation of Gymnema sylvestre-a multipurpose medicinal plant. Plant Cell Tiss. Org. Cult. 61: 97-105
Kumar, V., B. Jones and M. R. Davey. 1991. Transformation by Agrobacterium rhizogenes and regeneration of transgenic shoots of the wild soybean Glycine argyrea. Plant Cell Rep. 10:135-138.
Lane, W. D. 1979. In vitro propagation of Spirea bumalda and Prunus oistena from apices. Can. J. Plant Sci. 50:1025-1029.
Lee, Y. L., A. P. Sagare, C. Y. Lee, J. F. Shaw and H. S. Tsay. 2001. Formation of protoberberine-type alkaloids by the tubers of somatic embryo-derived plants of Corydalis yanhusuo. Planta Med. 67:839-842.
Li, H., S. J. Murch and P. K. Saxena, 2000. Thidiazuron-induced de novo shoot organogenesis on seedlings, etiolated hypocotyls and stem segments of Huang-qin. Plant Cell Tiss. Org. Cult. 62: 169-173.
Lloyd, G. amd B. McCown. 1980. Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip culture. Inti. Plant Prop. Soc. 30:421-427.
Lu, C. Y. 1993. The use of thidiazuron in tissue culture. In Vitro Cell Dev. Biol. Plant. 29:92-96.
Maeda, Y., Y. Fujitaand and Y. Yamada. 1983. Callus formation from protoplasts of cultured Lithospermum erythrorhizon cells. Plant Cell Rep. 2:179-182
Mantell, S. H., S. Q. Haque and A. P. Whitehall. 1978. Clonal propagation of Dioscorea alata L., Dioscorea rotundata Poir. yams by tissue culture. Hort. Sci. 53:95-98.
Mao, A. H., A. Wetten, M. Fay and P. D. S. Caligari. 1995. In vitro propagation of Clerodendrum colebrookianum Walp., a potentialnatural anti-hypertension medicinal plant. Plant Cell Rep. 14:493—496.
Mischenko, N. P., S. A. Fedoreyev, G. V. P. lazunov, G. K. Chernoded, V. P. Bulgakov and Y. N. Zhuravlev. 1999. Anthraquinone production by callus cultures of Rubia cordifolia. Fitoterapia 70:552-557
Miura, Y., K. Hirata, N. Kurano, K. Miyamoto and K. Uchida. 1988. Formation of vinblastine in multiple shoot culture of Catharanthus roseus. Planta Med. 54:18-20.
Moyano, E., S. Fornale, J. Palazon, R. M. Cusido, M. Bonfill, C. Morales and M. T. Pinol. 1999. Effect of Agrobacterium rhizogenes T-DNA on alkaloid production in Solanaceae plants. Phytochemistry 52:1287-1292.
Murashige, T. 1974. Plant propagation through tissue culture. Annu. Rev. Plant Physiol. 25:135-166.
Murashige, T. 1977. Manipulation of orange initiation in plant tissue culture. Bot. Bull. Acad. Sin. 18:1-24
Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15:437-497.
Nakagawa, K., H. Fukui and M. Tabata. 1986. Hormonal regulation of berberine production in cell suspension cultures of Thalictrum minus. Plant Cell Rep. 5:69-71.
Nigra, H. M., O. H. Caro and A. M. Giulietti. 1987. Production of solasodine by calli from different parts of Solanum eleagnifolium Cav. plant. Plant Cell Rep. 6:135-137.
Nishioka, I. 1988. Clonal multiplication of medicinal plant by tissue culture. Japan. J. Pharmac. 42:1-11.
Noboru, H., T. Kodamaand, and Y. Tomita. 1982. In Vitro propagation of Bupleurum falcatum. Shouyakugaku Zasshi 37:62-67.
Oksman-Caldentey, K. M. and R. Hiltunen. 1996. Transgenic crops for improved pharmaceutical products. Field Crops Res. 45:57-69
Parr, A. J., A. C. Peerless, J. D. Walton, R. J. Robins and M. J. C. Rhodes. 1988. Alkaloid production by transformed root culture of Catharanthus roseus. Plant Cell Rep. 7:309-312.
Patra, A., B. Rai, G. R. Rout and P. Das. 1998. Successful plant regeneration from callus cultures of Centella asiatica (Linn.) Urban. Plant Growth Regul. 24:13—16.
Pawlicki-Jullian, N., M. Sedira, and M. Welander. 2002. The use of Agrobacterium rhizogenes transformed roots to obtain transgenic shoots of the apple rootstock Jork 9. Plant Cell Tiss. Org. Cult. 70:163—171.
Richard, W., K. R. Pate and T. A. Thorpe. 1988. Ascorbic acid enhancement of organogensis in tabacco callus. Plant Cell Tiss. Org. Cult. 13:219-228.
Robb, S. M. 1957. The culture of excised tissue from bulb scales of Lilium speciosum Thum. J. Exp. Bot. 8:384-352.
Rout, G. R., S. Samantaray and P. Das. 2000. In vitro manipulation and propagation of medicinal plants. Biotechnol. Adv. 18:91-120.
Rout, G. R., C. Saxena, S. Samantaray and P. Das.1999. Rapid clonal propagation of Plumbago zeylanica Linn. Plant Growth Regul. 28:1—4.
Sahoo, Y. and P. K. Chand, 1998. Micropropagation of Vitex negundo L., a woody aromatic medicinal shrub, through high-frequency axillary shoot proliferation. Plant Cell Rep. 18:301-307.
Sahoo, Y., S. K. Pattnaik and P. K. Chand. 1997. In vitro clonal propagation of an aromatic medicinal herb Ocimum basilicum L.(Sweet basil) by axillary shoot proliferation. In Vitro Cell Dev. Biol. Plant 33:293—296.
Sanchez, M. I. 1988. Micropropagation of Cytopodium (Orchidaceae) through root tip culture. Lindleyana 3:93-96.
Sagare, A. P., C. L. Kuo, F. S. Chueh, and H. S. Tsay. 2001. De Novo Regeneration of Scrophularia yoshimurae YAMAZAKI (Scrophulariaceae) and quantitative Analysis of harpagoside, an iridoid glucoside, formed in aerial and underground parts of In vitro propagated and wild plants by HPLC. Biol. Pharm. Bull. 24：1311-1315.
Sasse, F., U. Heckenberg and J. Berlin. 1982. Accumulation of b-carbolin alkaloid and serotonin by cell culture of Peganum harmala L. I. Correlation between plants and cell cultures and influence of medium constituents. Plant Physiol. 69:400-404.
Sauerwein, M. and H. Becker. 1989. Vitamin B promotes growth of liverwort tissue culture. Agricell Rep. 12:28-30.
Saxena, C., G. R. Rout and P. Das. 1998. Micropropagation of Psoralea corylifolia Linn. J. Med. Aromatic Plant Sci. 20:15—18.
Sharma, T. R. and B. M. Singh. 1997. High frequency in vitro multiplication of disease-free Zingiber officinale Rosc. Plant Cell Rep. 17:68—72.
Shepard, J. F. and H. E. Street. 1974. The decline of embryogenic potential as callus and suspension cultures of carrot (Daucus carota L.) are serially subcultured. Ann. Bot. 38:223-241.
Shiau, Y. J., U. C. Chen, S. R. Yang, A. P. Sagare and H. S. Tsay. 2002. Conservation of Anoectochilus formosanus, a medicinally important terrestrial orchid, by synchronizing flowering, hand-pollination, and in vitro culture of seeds. Bot. Bull. Acad. Sin. 43:123-130.
Shimomura, K., K. Yoshimatsu, M. Jaziri and K. Ishimaru. 1997. Traditional medicinal plant genetic resources and biotechnology applications. In: Plant Biotechnology and Plant Genetic Resources for Sustainability and Productivity, eds. K. Watanabe and E. R. G. Pehu, pp. 209-225. Austin, Texas, R. G. Lands Company and Academic Press Inc.
Skoog, F. and C. O. Miller. 1957. Chemical regulation of growth and organ formation in plant tissue cultured in vitro. Symp. Soc. Exp. Biol. 11:118-131.
Smith, D. L. and A. D. Krikorian. 1990. Somatic proembryo production from excised, wounded zygotic carrot embryos on hormone-free medium: evaluation of the effects of pH, ethylene and activated charcoal. Plant Cell Rep. 9:34-37.
Stockigt, J., P. Obitz, H. Falkenhagen, R. Lutterbach and S.endreb. 1995. Natural products and enzymes from plant cell culture. Plant Cell, Tissue Organ Cult. 43:97-109.
Subroto, M. A., N. Artanti, D. Sudrajat, A. Djanakum and E. Widayat. 2001. Agrobacterium rhizogenes-mediated transformation of Solanum nigrum L.: spontaneous plant regeneration and endogenous IAA contents. Indon. J. Agri. Sci. 1: 53-59.
Touno, K., K. Harada, K. Yoshimatsu, K. Yazaki and K. Shimomura. 2000. Shikonin derivative formation on the stem of cultured shoots in Lithospermum erythrorhizon. Plant Cell Rep. 19:1121-1126.
Tsay, H. S., T. G. Gau and C. C. Chen. 1989. Rapid clonal propagation of Pinella ternata by tissue culture. Plant Cell Rep. 8:450-454.
Tsay, H. S. and H. L. Huang. 1997. Immature embryo culture of Angelica sinensis (Oliv.) Diels. Acta Hort. 447:207-209.
Verpoorte, R. and A. W. Alfermann. 2000. Plant secondary metabolism. In: Metabolic engineering of plant secondary metabolism, eds. Verpoorte, R, pp. 1-29. London, Kluwer academic publishers,
Vieira, R. F. and L. A. Skorupa. 1993. Brazilian medicinal plant gene bank. Acta Hort. 330:51-58.
Webb, K. J. and H. E.Street. 1977. Morphogenesis in vitro of Pinus and Picea. Acta Hort. 78:259—269.
Wink, M. 1990. Physiology of secondary product formation in plants. In: Secondary Products from Plant Tissue Culture, eds. B. V. Charlwood and M.J.C. Rhodesp, pp.23-41. New York, Oxford University Press.
Xu, X. H. and M. R. Davey. 1983. Shoot regeneration from mesophyll protoplasts and leaf explents of Rehmannia glutinosa. Plant Cell Rep. 2:55-57.
Yamamoto, H., N. Cohnyasu, K. Watanabe and T. Tomimori. 1986. Effects of carbon source on the growth and flavonoid formation of Scutellaria baicalensis stem callus cultures. Japan. J. Pharmac. 40:19-25.
Yamamoto, H., A. Kitayama and T. Tomimori. 1985. Root differentiation and paeoniflor in Paeonia lactiflora callus cultures. Japan. J. Pharmac. 39:185-189.
Yamamoto, O. and Y. Yamada. 1986. Production of reserpine and its optimization in cultured Rauwolfia serpentina Benth. cells. Plant Cell Rep. 5:50-53.
Yang, D. C. and Y. E. Choi. 2000. Production of transgenic plants via Agrobacterium rhizogenes-mediated transformation of Panax ginseng. Plant Cell Rep. 19:491-496.
Yoshikawa, T. and T. Furuya. 1987. Saponin production by cultures of Panax ginseng transformed with Agrobacterium rhizogenes. Plant Cell Rep. 6:449-453.
Zhang, S. and K. C. Cheng. 1988. Angelica sinensis (OLIV) DIELS : In vitro culture, regeneration and the production of medicinal compounds. In: Biotechnology in Agriculture and Forestry, ed. B. Y. P. S. Bajaj, pp. 1-22. Springer-Verlag.