Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22870
標題: 建立蜜糖文心蘭體胚繁殖系統及ZmCCD1基因轉殖至南西文心蘭之探討
Somatic embryogenesis of Oncidium Sweet Sugar and transformation of maize carotenoid cleavage dioxygenase 1 gene into Oncidium Gower Ramsey
作者: 鄭郁琪
Cheng, yu-chi
關鍵字: 文心蘭
thidiazuron
擬原球體
農桿菌
類胡蘿蔔素
香味基因
Carotnoid Cleavage Dioxygenase 1 (CCD1)
apocarotenoid
Agrobacterium transformation of Oncidium NDM medium
protocorm like bodies (PLBs)
出版社: 生命科學系所
引用: 何松林 (2001) 不同基本培養基及培養方式對文心蘭原球莖增殖的影響. 華北農學報 18, 88-91. 何慧敏 (2005) 蝴蝶蘭花粉與農桿菌共培養基因轉殖系統之建立. 朝陽科技大學生物技術研究所碩士論文. 李淑華 (1999) 文心蘭體胚分化及基因轉殖系統之研究. 國立成功大學碩士論文. 林金其 (1998) 蝴蝶蘭花梗結間培養之擬圓球體誘導再生過程型態學. 洋蘭月刊 20, 85-89. 邱鴻毅 (1998) 蝴蝶蘭基因轉殖最佳條件之研究. 國立成功大學研究所碩士論文. 姚麗娟, 徐曉薇, 林紹生, 游聚斌, 陳中林 (2004) 文心蘭莖尖及花梗離體培養研究. 福建熱作科技 29, 5-6. 楊淑蓉 (2003) 文心蘭以農桿菌媒介基因轉殖之研究. 高雄師範大學生物科學研究所 詹明才 (1992) 農桿菌轉殖水稻系統的建立. 國立台灣大學農藝學研究所博士論文. 潘學鋒, 王日暖, 莫海 (2001) 文心蘭莖尖離體培養研究. 熱帶林業 29, 145-157. Auldrige, M.E., McCarty, D.R., and J., K.H. (2006a) Plant Carotenoid cleavage oxygenase and their apocarotenoid products. Current Opinion in Plant Biology 9, 315-321. Auldrige, M.E., Block, A., Vogel, J.T., Dabney-Smith, C., Mila, I., and Bouzayen, M. (2006b) Characterization of three members of the Arabidopsis carotenoid cleavage dioxygenase family demonstrates the divergent roles of this multifuctional enzyme family. Plant J. 45, 982-983. Belarmino, M.M., and Mii, M. (2000) Agrobacterium-mediated genetic transformation of a phalaenopsis orchid. Plant Cell Rep. 19, 435-442. Bouvier, F., Isner, J.C., Dogbo, O., and Camara, B. (2005) Oxidative tailoring of carotenoid: a prospect towards novel functions in plants. Trend Plant Sci. 10, 187-194. Camara, B., and Bouvier, F. (2004) Oxidative remodeling of plastid carotenoids. Archives of Biochemistry and Biophysics 430 (2004) 16-21 430, 16-21. Chai, M.L., Xu, C.J., Senthil, K.K., kim, J.Y., and kim, D.H. (2002) Stable transformation of protocorm-like bodies in Phalaenopsis orchid mediated by Agrobacterium tumefaciens. Sci. Horic. 96, 11. Chan, M.T., and Silva, J.A.T.d. (2006) Oncidium tissue culture, transgenic and biotechonology. In Biotechonology:advances and tropical issues (London,UK: Global Science Books, Ltd), pp. 193-198. Chen, J.T., and Chang, W.C. (2000) Plant regeneration via embryo and shoot bud formation from flower-stalk explants of Oncidium Sweet Sugar. Plant Cell ,Tissue and Organ Cult. 62, 95-100. Cheung, A.Y., McNellis, T., and Piekos, B. (1993) Maintenance of chloroplast compoonents during chromoplast differentiation in the tomato mutant Green Flesh. Plant Physiol. 101, 1223-1229. Chin, D.P., Mishiba, K.-i., and Mii, M. (2007) Agrobacterium-mediated transformation of protocorm-like bodies in cymbidium. Plant Cell Rep. Vol. 26, 735-743. Culter, A.J., and Krochko, J.E. (1999) Formation and breakdown of ABA. Trend Plant Sci. 4, 472-478. Cunningham, F.X. (2002) Regulation of carotenoid synthesis and accumulation in plants. Pure Appl. Chem. 74, 1409-1417. Cunningham, F.X., and Grantt, E. (1998) Genes and enzymes of carotenoid biosynthesis in plants. Annu. Rev. Plant Physiol. 49, 557-583. Fester, T. (1999) Accumulation of secondary compounds in barley and wheat roots in response to inculation with an arbuscular mycorrhizal fungus and co-inoculation with rhizosphere bacteria. Mycorrhiza 8, 241-246. Floss, D.S., and Walter, M.H. (2009) Role of carotenoid cleavage dioxygenase 1 (CCD1) in apocarotenoid biogenesis revised. Plant signaling & Behavior 4, 172-175. Floss, D.S., Schliemann, W., Schmidt, J., Strack, D., and Walter, M.H. (2008) RNA-intererence-mediated repression of MtCCD1 in mycorrhizal roots of Medicago truncatula causes accumulation of C27 apocarotenoids, shedding light on the functional role of CCD1. Plant Physiol. 148, 1267-1282. Fray, R.G., Wallace, A., Fraser, P.D., Valero, D., Hedden, P., and Bramley, P.M.G., D. (1995) Constitutive expression of a fruit phytoene synthase gene in transgenic tomatoes causes dwarfism by redirectin metabolites from the gibberellin pathway. Plant J. 8, 693-701. Giuliano, G., Al-Bili, S., and Lintig, J.v. (2003) Carotenoid oxygenases: cleave it or leave it. Trend Plant Sci., 145-148. Hong, P.I., J.T.Chen, and Chang, W.C. (2008) Promotion of direct somatic embryogenesis of Oncidium by adjustin carbon sources. Biologia Plantarum 52, 3. Howitt, C.A., and Pogson, B.J. (2006) Carotenoid accumulation and function in seeds and non-green tissues. Plant Cell Environ 29, 435-445. Islam, M.O., Rahman, A.R.M.M., Matsui, S., and Prodhan, A.K.M.A.-u.-d. (2003) Effects of complex organic extracts on callus growth and PLB regeneration through embryogenesis in the Doritaenopsis orchid. J.A.R.Q. 37, 229-235. Keppler, L.D., Baker, C.J., and Atkinson, M.M. (1989) Active oxygem production during bacteria-induced hypersensitive reaction in tobacco suspension cells. Phytopathology 79, 974-978. Klingner, A., Hundeshagen, B., Kernebeck, H., and Bothe, H. (1995) Localization of the yellow pigment formed in roots of gramineous plants colonized by arbuscular fungi. Protoplasma 185, 50-57. Ku, M.S.B., Agarie, S., Nomura, M., Funkayama, H., and Tsuchida, H. (1999) High level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants. Nat. Biotehnol. 17, 76-80. Levin, A., Tenhaken, R., Dixon, R., and Lamb, C. (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79, 583-593. Liau, C.-H., You, S., Prasad, V., Hsiao, H., JC, L., NS, Y., and MT, C. (2003) Agrobacterium tumefaciens-mediated transformation of an Oncidium orchid. Plant Cell rep. 21, 993-998. Liu, T.-H., Lin, J.-J., and Wu, R.-Y. (2006) The Effects of Using Trehalose as a Carbon Source on the Proliferation of Phalaenopsis and Doritaenopsis Protocorm-Like-Bodies. Plant Cell, Tissue and Organ Culture 86, 125-129. Mouchel, C.F., and Leyser, O. (2007) Novel phytohormomes involved in long-range signaling. . Current opinion in plant Biology 10, 473-476. Ohmiya, A. (2009) Carotenoid cleavage dioxygenase and their apocarotenoid products in plants. Plant Biotechnology 26, 351-358. Roy, J., and Banerjee, N. (2003) Induction of callus and plant regeneration from shoot-tip explants of Dendrobium fimbriatum Lindl. var. oculatum Hk. f. Sci. Horic. 97, 333-340. Schlichr, M., Samajova, O., Schachtschabel, D., Mancuso, S., Menzel, D., and Boland, W. (2008) D''orenone blocks polarized tip growth of root hairs by interfering with the PIN2-mediated auxin transport network in the root apex. Plant J. 55, 709-717. Schwartz, S.H., Qin, X., and Zeevaart, J.A.D. (2001) Characterization of a Novel Carotenoid Cleavage Dioxygenase from plants. J. Biol. Chem. 276, 25208-25211. Schwartz, S.H., Tan, B., C., Gage, D.A., Zeevaart, J.A., and McCarty, D.R. (1997) Specific oxidative cleavage of carotenoids by VP14 of maize. . Science 276, 1872-1874. Silva, J.A.T.d., Chan, M.T., Sanjaya, Chai, M.L., and Tanaka, M. (2006) Priming abiotic factors for optimal hybrid Cymbidium (Orchidaceae) PLB and callus induction, plantlet formation,and their subsequent cytogenetic stability analysis. Sci. Horic. 109, 368-378. Simkin, A.J., Schwartz, S.H., Auldrige, M.E., Tayler, M.J., and Klee, H.J. (2004) The tomato carotenoid cleavage dioxygenase controls emission of β- inonem pseudionone and geranylacetone. Plant J. 40, 882-892. Sun, Z., Hans, J., Walter, M.H., Matusova, R., Beekwilder, J., Verstappen, G.W.A., Ming, Z., Eschtelt, E.v., Dieter, S., Bisseling, T., and Bouwmeester, H.J. (2008) Cloning and characterisation of a maize carotenoid cleavage dioxygenase (ZmCCD1) and its involvement in biosynthesis of apocarptenoids with various roles in mutualistic and parasitic interactions. Planta 228, 789-801. Tokuhara, K., and Mii, M. (1993) Micropropagation of Phalaenopsis and Doritaenopsis by culturing shoot tips of flower stalk buds. Plant Cell Rep. 13, 7-11. Tokuhara, K., and Mii, M. (2003) Highly-efficient somatic embryogenesis from cell suspension cultures of phalaenopsis orchids by adjusting carbohydrate sources. In Vitro Cellular & Developmental Biology - Plant 39, 635-639. Vernade, D., Herrera-Estrella, A., Wang, K., and Van Montagu, M. (1988) Glycine betaine allows enhanced induction of the Agrobacterium tumefaciens vir genes by acetosyringone at low pH. J. Bacteriol. 170, 5822-5829. Vogel, J.T., Tan, B., C., McCarty, D.r., and Klee, H.J. (2008) Carotenoid cleavage dioxygenase 1 enzyme has broad substrate specificity, cleaving multiple carotenoids at two different bond positions. J. Biol. Chem. 283, 11364-11373. Walter, M.H., Fester, T., and Strack, D. (2000) Arbuscular mycorrhizal fungi induce the non-mevalonate methlerythritol phosphate pathway of isoprenoid biosynthesis correlated with the accumulation of the "yellow pigment" and other apocarotenoids. Plant J. 21, 571-578. Winterhalter, P., and Rouseff, R.L. (2002) Carotenoid-derived aroma compounds. (American Chemical Society ), pp. 1-17. Yu, H.Y., S.H., and Goh, C.J. (2001) Agrobacterium-mediated transformation of a Dendrobium orchid using the class 1 knox gene DOH1. Plant Cell Rep. 20, 301-305.
摘要: 文心蘭Oncidium Sweet Sugar及Gower Ramsey商業價值高,但前者無法快速量產,後者缺乏香氣。本研究初步評估農桿菌轉殖技術對Gower Ramsey花香及Sweet Sugar產量之改善效果,以期增加此兩者之產業效益。 文心蘭Oncidium Sweet Sugar屬於盆花品系,目前商業為成熟株側芽增殖方式量產。本實驗建立擬原球體(protocorm-like bodies , PLBs)繁殖系統,期望能在幼年期即可提高其產率。以細胞分裂素BA (benzoic acid)、TDZ (thidiazuron)以及生長素NAA (naphthaleneacetic acid)搭配不同的培養基與醣類,誘導培植體由癒合組織分化成PLB。在BA /NAA=10的比例,BA為1 mg/L時,最適合誘導莖頂癒合組織。液態的誘導系統雖然褐變率高,但是誘導成功的培植體可快速增殖出大量的癒合組織。單獨使用TDZ易使植株褐變;但0.5 mg/L 的TDZ搭配0.5 mg/L的BA適合誘導癒合組織。 單獨添加1 mg/L BA或以0.5 mg/L BA搭配 1 mg/L TDZ適合PLBs分化。 0.5 mg/L 的TDZ添加在NAA則易使植株不經由PLBs途徑直接分化成植株。基本培養基NDM (New Dogashima Medium) 添加蔗糖的基本培養基最適合Sweet Sugar原球體增殖。 Sweet Sugar農桿菌轉殖系統測試實驗,在液態MS培養基中將農桿菌及癒合組織共培三天後除菌,經過一個月以上的復原期後,克服農桿菌引起之過敏反應 (hypersensitive response),再以非hygromycin的抗生素篩選二個月以上,才能初步篩選轉殖株。 目前已知文心蘭Gower Ramsey唇瓣含有豐富的類胡蘿蔔素,且類胡蘿蔔素和玉米的carotenoid cleavage dioxygenase 1 (ZmCCD1)作用可產生香氣,我們假設藉由農桿菌送入的ZmCCD1可和Gower Ramsey內的類胡蘿蔔素結合並產生花香,並藉由PCR與GUS染色法鑑定轉殖成功,初步確認外源基因已確實進入轉殖株。未來觀察將著重於HPLC或GC-MS分析ZmCCD1 參與文心蘭花瓣中何種類胡蘿蔔素代謝路徑,影響胡蘿蔔素的代謝,改變下游文心蘭的花色,產生帶有白花並帶有香氣的文心蘭新品系。
Oncidium ‘Sweet Sugar'' has economical importance in pot-plant industries recently but its micropropagation has not been well-studied. This study aims to investigate Agrobacterium tumefaciens mediated gene transformation of ‘Sweet Sugar'. Recently, studies on Oncidium' Gower Ranmsey' floral color which composed of carotenoid have been revised. However, there are rare research articles have been published as synchronal remodeling floral color and aromatic compounds of Oncidium cultivars. In this study, in vitro callus and protocorm-like bodies (PLBs) propagation system had been established. Shoot tip cultured in NDM medium was supplied with 1mg/L BA and 0.1 mg/L NAA which was the best formula for somatic embryogenesis. Only TDZ showed not suitable for somatic embryogenesis induction on NDM medium. For embryogenesis differentiation, adding 0.5 mg/L BA on NDM medium supplied with TDZ preferred to PLBs formation. Subculture 0.5 mg/L BA with 1mg/L TDZ proliferated the best quantity of PLBs. In this research, 0.5 mg/L NAA supplied with TDZ in NDM medium induced embryonic callus differentiation to shoot buds directly. This study developed Oncidium' Sweet Sugar' with Agrobacterium mediated gene transformation system preliminarily. In transformation step, co-cultured Agrobacterium with Oncidium 'Sweet Sugar” callus in liquid MS medium was better than in solid MS medium. After transformation, over 1-month recovering time is required to overcome the hypersensitive response, which was triggered by bacterial infection. It is necessary to transfer the survival explants and placed on selection medium for at least 2 months, in order to confirm the transgenosis plants. This research transformed Oncidium 'Gower Ramsey' PLBs with Agrobacterium which carrying maize carotenoid cleavage dioxygenase genes (ZmCCD1). The transgenic PLBs were confirmed by PCR and GUS histochemical staining. These results demonstrated that ZmCCD1 can be transferred in Orchid successfully. It is anticipated that transformed ZmCCD1 in Oncidium will elevate flower aroma and change the color of Orchid. We need to wait for transgenic plants to bloom, in order to accomplish further study as analyzing aroma volatiles and colors of flowers by using HPLC and GC-MS.
URI: http://hdl.handle.net/11455/22870
其他識別: U0005-0802201019545400
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-0802201019545400
Appears in Collections:生命科學系所

文件中的檔案:

取得全文請前往華藝線上圖書館



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.