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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 杜宜殷(Yi-Yin Do) | |
dc.contributor.author | Chih-Fan Wang | en |
dc.contributor.author | 汪芝帆 | zh_TW |
dc.date.accessioned | 2021-06-16T02:30:40Z | - |
dc.date.available | 2020-08-31 | |
dc.date.copyright | 2015-08-31 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-07-30 | |
dc.identifier.citation | 胡慧琳. 2012. 香蕉乙烯訊息傳導MhCTR1基因之表現及啟動子活性分析. 國立臺灣大學園藝暨景觀學系碩士論文. 張家瑋. 2011.嘉德麗雅蘭再生及轉殖系統建立. 國立臺灣大學園藝暨景觀學系碩士論文. 程永煒. 2012. 應用轉位子系統去除轉殖蝴蝶蘭篩選標誌基因之研究. 國立臺灣大學園藝暨景觀學系碩士論文. 衛宛君. 2013. 應用轉位子去除篩選標誌於基因轉殖植物抗東亞蘭嵌紋病毒及齒舌蘭輪點病毒之研究. 國立臺灣大學園藝暨景觀學系碩士論文. Baulcombe, D. 2004. RNA silencing in plants. Nature 431: 356-363. Beaudoin, N., C. Serizet, F. Gosti, and J. Giraudat. 2000. Interaction between abscisic acid and ethylene signaling cascades. Plant Cell 12:1103-1115. Ben-Haj-Salah., H. and F. Tardieu. 1995. Temperature affects expansion rate of maize leaves without change in spatial distribution of cell length. Plant Physiol. 109:861-870. Beranova, M., S. Rakousky, Z. Vavrova, and T. Skalicky. 2008. Sonication assisted Agrobacterium-mediated transformation enhances the transformation efficiency in flax (Linum usitatissimum L.). Plant Cell Tiss. Organ Cult. 94:253-259 Bleecker, A.B., M.A. Estelle, C. Somerville, and H. Kende. 1988. Insensitivity to ethylene conferred by a dominant mutation in Arabidopsis thaliana. Science 241:1086-1089. Bovy, A.G., G.C. Angenent, H.J.M. Dons and A.C. van Altvorst. 1999. Heterologous expression of the Arabidopsis etr1-1 allele inhibits the senescence of carnation flowers. Mol. Breed. 5:301-308. Chai, M.L., C.J. Xua, K.K. Senthil, J.Y. Kim, and D.H. Kim. 2002. Stable transformation of protocorm-like bodies in Phalaenopsis orchid mediated by Agrobacterium tumefaciens. Sci. Hortic. 96:213-224 Chang, C., S.F. Kwok, A.B. Bleecker, and E.M. Meyerowitz. 1993. Arabidopsis ethylene-response gene ETR1: similarity of product to two-component regulators. Science 262:539-544. Chang, H., M.L. Jones, G.M. Banowetz, and D.G. Clark. 2003. Overproduction of cytokinins in Petunia flowers transformed with PSAG12-IPT delays corolla senescence and decreases sensitivity to ethylene. Plant Physiol. 132:2174-2183. Chen, M.K., W.H. Hsu, P.F. Lee, M. Thiruvengadam, H.I. Chen, and C.H. Yang. 2011. The MADS box gene, FOREVER YOUNG FLOWER, acts as a repressor controlling floral organ senescence and abscission in Arabidopsis. Plant J. 68:168-185. Chen, T.Y., J.T. Chen, and W.C. Chang. 2004. Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell Tiss. Organ Cult.76:11-15. Chia, T.F., Y.S. Chen, and N.H. Chua. 1992. Characterization of Cymbidium mosaic virus coat protein gene and its expression in transgenic tobacco plants. Plant. Mol. Biol. 18;1091-1099. Christensen, B. and R. Muller. 2009. Kalanchoe blossfeldiana transformed with rol genes exhibits improved postharvest performance and increased ethylene tolerance. Postharvest Biol. Technol. 51:399-406. Cueva Agila, A.Y., I. Guachizaca, and R. Cella. 2013. Combination of 2, 4-D and stress improves indirect somatic embryogenesis in Cattleya maxima Lindl. Plant Bio. doi: 10.1080/11263504.2013.797033. Day, A.G., E.R. Bejarano, K.W. Buck, M. Burrell, and C.P. Lichtenstein. 1991. Expression of an antisense viral gene in transgenic tobacco confers resistance to the DNA virus tomato golden mosaic virus. Proc. Natl. Acad. Sci. USA. 88:6721-6725. Dellaporta, S.L., J. Wood, and J.B. Hicks. 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep. 1:19-21. Feinberg, A.P. and B. Vogelstein. 1983. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132:6-13. Gan, S. and R.M. Amasino. 1995. Inhibition of leaf senescence by autoregulated production of cytokinin. Science 270:1986-1988 Gao, Z., Y.F. Chen, M.D. Randlett, X.C. Zhao, J.L. Findell, J.J. Kieber, and G.E. Schaller. 2003. Localization of the Raf-like kinase CTR1 to the endoplasmic reticulum of Arabidopsis through participation in ethylene receptor signaling complexes. J. Biol. Chem. 278:34725-34732. Ghassemian, M., E. Nambara, S. Cutler, H. Kawaide, Y. Kamiya, and P. McCourt. 2000. Regulation of abscisic acid signaling by the ethylene response pathway in Arabidopsis. Plant Cell 12:1117-1126 Goh, C.J., A.H. Halevy, R. Engel, and A.M. Kofranek. 1985. Ethylene evolution and sensitivity in cut orchid flowers. Sci Hortic. 26: 57-67. Gomez-Jimenez, M.C., E. Garcia-Olivares, and A.J. Matilla. 2001. 1-Aminocyclopropane-1-carboxylate oxidase from embryonic axes of germinating chick-pea (Cicer arietinum L.) seeds: cellular immunolocalization and alterations in its expression by indole-3-acetic acid, abscisic acid and spermine. Seed Sci. Res. 11:243–253. Gonzalez, E.R., A. Andrade, A.L. Bertolo, G.C. Lacerda, R.T. Carneiro, V. A.P. Defavari, M.T.V. Labate, and C.A. Labate. 2002. Production of transgenic Eucalyptus grandis × E. urophylla using the sonication-assisted Agrobacterium transformation (SAAT) system. Funct. Plant Biol. 29:97-102. Gow, W.P., J.T. Chen, and W.C. Chang. 2010. Enhancement of direct somatic embryogenesis and plantlet growth from leaf explants of Phalaenopsis by adjusting culture period and explants length. Acta Phsiol. Plant 32:621-627. Hiei, Y., S. Ohta, T. Komari, and T. Kumashiro. 1994. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6: 271-282. Hoshi, Y., M. Kondo, S. Mori, Y. Adachi, M. Nakano, and H. Kobayashi. 2004. Production of transgenic lily plants by Agrobacterium-mediated transformation. Plant Cell Rep. 22:359-364. Huang, L.C., U.L. Lai, S.F. Yang, M.J. Chu, C.I. Kuo , M.F. Tsai, and C.W. Sun. 2007. Delayed flower senescence of Petunia hybrida plants transformed with antisense broccoli ACC synthase and ACC oxidase genes. Postharvest Biol. Technol. 46:47-53. Huang, Y., H. Li, C.E. Hutchison, J. Laskey, and J.J. Kieber. 2003. Biochemical and functional analysis of CTR1, a protein kinase that negatively regulates ethylene signaling in Arabidopsis. Plant J. 33:221-233. Isomura, Y., Y. Matumoto, A. Murayama, M. Chatani, N. Inouye, and M. Ikegami. 1991. Molecular cloning, sequencing and expressing in Escherichia coli of the Odontoglossum ringspot virus coat protein gene. J. Gen.Virol. 72:2247-2249. Jin, S., X. Zhang, S. Liang, Y. Nie, X. Guo, and C. Huang. 2005. Factors affecting transformation efficiency of embryogenic callus of Upland cotton (Gossypium hirsutum) with Agrobacterium tumefaciens. Plant Cell Tiss. Organ Cult. 81:229-237. Jorgensen, R.A., P.D. Cluster, J. English, Q. Que, and C.A. Napoli. 1996. Chalcone synthase cosuppression phenotypes in petunia flowers: comparison of sense vs. antisense constructs and single-copy vs. complex T-DNA sequences. Plant Mol. Bio. 31: 957-973. Khachatourians, G.G., A. McHughen, R. Scorza, W.K. Nip, and Y.H. Hui. 2002. Transgenic linseed flax, p. 870-886. In: Transgenic plants and crops. Marcel Dekker, Inc. N.Y. Kieber, J.J., M. Rothenberg, G. Roman, K.A. Feldmann, and J.R. Ecker. 1993. CTR1, a negative regulator of the ethylene response pathway in Arabidopsis, encodes a member of the raf family of protein kinases. Cell 72:427-441. Kim, J.D., Y.B. Koo, and M.U. Chang. 1998. Genome characterization of a Korean isolate of Cymbidium mosaic virus. Mol. Cells 30:181-188. Kim, S.Y., C.B. Hong, and I. Lee. 2001. Heat shock stress causes stage-specific male sterility in Arabidopsis thaliana. J. Plant Res. 114: 301-307. Knudson, L. 1946. A new nutrient solution for germination of orchid seed. Am. Orchid Soc. Bull. 15: 214–217. Kuo, H.L., J.T. Chen, and W.C. Chang. 2005. Efficient plant regeneration through direct somatic embryogenesis from leaf explants of Phalaenopsis ‘Little Steve’. In Vitro Cell. Dev. Biol. 41:453-456. Larkindale, J., and B. Huang. 2004. Thermotolerance and antioxidant systems in Agrostis stolonifera: Involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene. J. Plant Physiol. 161: 405-413. Larkindale, J., and M.R. Knight. 2002. Protection against heat stress-induced oxidative damage in Arabidopsis involves calcium, abscisic acid, ethylene, and salicylic acid. Plant Physiol. 128: 682-695. Lawson, R.H. and M. Brannigan. 1986. Virus disease of orchids. Handbook on orchid pests and diseases. American Orchid Society, West Palm Beach, Forida. Lawton, K.A., B. Huang, P.B. Goldsbrough, and W.R. Woodson. 1989. Molecular cloning and characterization of senescence-related genes from carnation flower petals. Plant Physiol. 90:690-696. Leclercq, J., C. Lori, A. Phillips, H. Zegzouti, B. Jones, A. Latche, J.J. Giovannoni, J. Pech, and M. Bouzayen. 2002. LeCTR1, a tomato CTR1-like gene, demonstrates ethylene signaling ability in Arabidopsis and novel expression patterns in tomato. Plant Physiol. 130:1132-1142. Lee, Y.I. and N. Lee. 2003. Plant regeneration from protocorm-derived callus of Cypripedium formosanum. In Vitro Cell. Dev. Biol. 39:475-479. Liao, L.J., I.C. Pan, Y.L. Chan, Y.H. Hsu, W.H. Chen, and M.T. Chan. 2004. Transgene silencing in Phalaenopsis expressing the coat protein of Cymbidium Mosaic Virus is a manifestation of RNA-mediated resistance. Mol. Breed. 13:229-242. Linkies, A., K. Muller, K. Morris, V. Tureckova, M. Wenk, C.S.C. Cadman, F. Corbineau, M. Strnad, J.R. Lynn, W.E. Finch-Savage, and G. Leubner-Metzger. 2009. Ethylene interacts with abscisic acid to regulate endosperm rupture during germination: a comparative approach using Lepidium sativum and Arabidopsis thaliana. Plant Cell 21:3803-3822. Lopez, R.G. and E.S. Runkle. 2005. Environmental physiology of growth and flowering of orchids. HortScience 40:1969-1973. Marcotrigiano, M., S.E. McGlew, G. Hackett, and B. Chawla. 1996. Shoot regeneration from tissue-cultured leaves of the American cranberry ( Vaccinium macrocarpon). Plant Cell Tiss. Organ Cult. 44: 195-199. Matilla, A.J. 2000. Ethylene in seed formation and germination. Seed Sci. Res. 10:111–126. Mishiba, K.I., D.P. Chin, and M. Mii. 2005. Agrobacterium-mediated transformation of Phalaenopsis by targeting protocorms at an early stage after germination. Plant Cell Rep. 24:297–303 Muller, B., M. Stosser, and F. Tardieu. 1998. Spatial distributions of tissue expansion and cell division rates are related to irradiance and to sugar content in the growing zone of maize roots. Plant. Cell Environ. 21:149-158. Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant 15:473-497. Narinobu, I. 1983. Host range and properties of a strain of Odontoglossum ringspot virus in Japan. Nogaku Kenkyu. 60:53-57. Norelli, J., J. Mills, and H. Aldwinckle. 1996. Leaf wounding increases efficiency of Agrobacterium-mediated transformation of apple. HortScience 31:1026-1027. Patterson, S.E. and A.B. Bleecker. 2004. Ethylene-dependent and -independent processes associated with floral organ abscission in Arabidopsis. Plant Physiol. 134:194-203. Person, M.N. and J.S. Cole. 1991. Further observations on the effect of Cymbidium mosaic virus and Odontoglossum ringspot virus on the growth of Cymbidium orchids. J. Phytopath. 131:193-198. Pieterse, C.M.J., S.C.M. Wees, J.A. Pelt, M. Knoester, R. Laan, H. Gerrits, P.J. Weisbeek, and L.C. Loon. 1998. A novel signaling pathway controlling induced systemic resistance in Arabidopsis. Plant Cell 10:1571-1580. Raffeiner, B., M. Serek, and T. Winkelmann. 2009. Agrobacterium tumefaciens-mediated transformation of Oncidium and Odontoglossum orchid species with the ethylene receptor mutant gene etr1-1. Plant Cell Tiss. Organ Cult. 98:125–134. Rogers, H.J. 2013. From models to ornamentals: how is flower senescence regulated? Plant Mol. Biol. 82:563-574. Roy, J. and N. Banerjee. 2003. Induction of callus and plant regeneration from shoot-tip explants of Dendrobium fimbriatum Lindl. var. oculatum Hk. f. Scientia Horticulturae. 97:333-340. Schmulling, T., J. Schell, and A. Spena. 1988. Single genes from Agrobacterium rhizogenes influence plant development. EMBO J. 7:2621-2629. Shaw, J.F., H.H. Chen, M.F. Tsai, C.I. Kuo, and L.C. Huang. 2002. Extended flower longevity of Petunia hybrida plants transformed with boers, a mutated ERS gene of Brassica oleracea. Mol. Breed. 9:211-216. Shibuya, K., K.G. Barry, J.A. Ciardi, H.M. Loucas, B.A. Underwood, S. Nourizadeh, J.R. Ecker, H.J. Klee, and D.G. Clark. 2004. The central role of PhEIN2 in ethylene responses throughout plant development in Petunia. Plant Physiol. 136:2900-2912. Shrestha, B.R., D.P. Chin, K. Tokuhara, and M. Mii. 2007. Efficient production of transgenic plantls of Vanda through sonication-assisted Agrobacterium-mediated transformation of protocorm-like bodies. Plant Biotechnol. 24:429-434. Sjahril, R., D.P. Chin, R.S. Khan, S. Yamamura, I. Nakamura, Y. Amemiya, M. Mii. 2006. Transgenic Phalaenopsis plants with resistance to Erwinia carotovora produced by introducing wasabi defensin gene using Agrobacterium method. Plant Biotechnol. 23:191-194. Song, d R.F. Allison. 2013. Engineering cherry rootstocks with resistance to Prunus necrotic ring spot virus through RNAi-mediated silencing. Plant Biotechnol. J. 11:702-708. Southern, E.M. 1975. Deteection of specific sequence among DNA fragment separted by gel electrophoresis. J. Mol. Biol. 98:503-517. Teixeira, S. and A. Jaime. 2012. New basal media for protocorm-like body and callus induction of hybrid Cymbidium. Fruit Ornam. Plant Res. 20:127-133. Tougou, M., N. Yamagishi, N. Furutani, Y. Shizukawa, Y. Takahata, and S. Hidaka. 2007. Soybean dwarf virus-resistant transgenic soybeans with the sense coat protein gene. Plant Cell Rep. 26:1967-1975. Trick, H.N. and J.J. Finer. 1998. Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill] embryogenic suspension culture tissue. Plant Cell Rep. 17: 482-488. Trick, H. and J.J. Finer. 1997. SAAT: sonication-assisted Agrobacterium-mediated transformation. Transgenic Res. 6:329-336. Wang, H., G. Stier, J. Lin, G. Liu, and Z. Zhang. 2013. Transcriptome changes associated with delayed flower senescence on transgenic Petunia by inducing expression of etr1-1, a mutant ethylene receptor. PLoS ONE 8:e65800. Wilson, R.C. and J.A. Doudna. 2013. Molecular mechanisms of RNA interference. Ann. Rev. Biophys. 12:677-687. Wong, S.M., P.H. Mahtani, K.C. Lee, H.H. Yu, Y.Tan, K.K. Neo, Y. Chan, M. Wu, and C.G. Chang. 1997. Cymbidium misaic potexvirus RNA: complete nucleotide sequence and phylogenetic analysis. Arch. Virol. 142:383-391. Yadav, J.S., E. Ogwok, H. Wagaba, B.L. Patil, B. Bagewadi, T. Alicai, E. Gaitan-Solis, N.J. Taylor, and C.M. Fauquet. 2011. RNAi-mediated resistance to Cassava brown streak Uganda virus in transgenic cassava. Mol. Plant Pathol. 12:677-687. Yan, B., M.S.S. Reddy, G.B. Collins, and R.D. Dinkins. 2000. Agrobacterium tumefaciens-mediated transformation of soybean [Glycine max (L.) Merrill.] using immature zygotic cotyledon explants. Plant Cell Rep. 19:1090-1097. Zeng, S.J. 2011. Asymbiotic seed germination, induction of calli and protocorm-like bodies, and in vitro seedling development of the rare and endangered Nothodoritis zhejiangensis Chinese orchid. HortScience 46:460-46. Zettler, F.W., N.J. Ko, G.C. Wisler, M.S. Elliot, and S.M. Wong. 1990. Viruses of orchids and their control. Plant Dis. 74:621-626. Zhang, L., D.P. Chin, M. Fukami, H. Ichikawa, I. Nakamura, and M. Mii. 2010. Agrobacterium-mediated genetic transformation of Cattleya with an Odontoglossum ringspot virus replicase gene sequence. Plant Biotechnol. 27:421-426. Zhang, P., H. Vanderschuren, J. Fűtterer, and W. Gruissem. 2005. Resistance to cassava mosaic disease in transgenic cassava expressing antisense RNAs targeting virus replication genes. Plant Biotechnol. J. 3:385-397. Zhao, Z.Y., W. Gu, T. Cai, L. Tagliani, D. Hondred, D. Bond, S. Schroeder, M. Rudert, and D. Pierce. 2001. High throughput genetic transformation mediated by Agrobacterium tumefaciens in maize. Mol. Breed. 8:323-333. Zheng, Q., Y. Zheng, G. Wang, W. Guo, and Z. Zhang.2007. Sonication assisted Agrobacterium-mediated transformation of chalcone synthase (CHS) gene to Spring Dendrobium cultivar ‘Sanya’. Afr. J. Biotechnol. 10: 11832-11838. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/53825 | - |
dc.description.abstract | 嘉德麗雅蘭 (Cattleya spp.) 因對乙烯敏感,以致於花期不長。本研究欲利用生物技術抑制乙烯訊息傳導,以延長嘉德麗雅蘭之花期。以嘉德麗雅蘭葉片為材料,利用農桿菌媒介法,將過量表現香蕉之constitutive triple response 1 (Musa spp. Constitutive Triple Response 1, MhCTR1) 基因之質體轉殖至嘉德麗雅蘭中,測試條件包含預培養、農桿菌液濃度、超音波震盪時間及共培養天數。為驗證策略之可行性,將過量表現MhCTR1質體轉殖至阿拉伯芥。過量表現MhCTR1之T1代阿拉伯芥,與未轉殖株相比,有較大之花朵及較長之花期。過量表現MhCTR1之T3代阿拉伯芥種子,經ACC處理後,與未轉殖株相比並無明顯之乙烯三相反應,顯示此延緩老化之策略可有效抑制乙烯反應。嘉德麗雅蘭葉片經15秒超音波震盪及暗培養2週之處理,為最適預培養條件;以低農桿菌液濃度進行轉殖,有較高之芽體再生率及較低之農桿菌復發率。抗病基因轉殖方面,係利用核糖核酸干擾技術,將同時含蕙蘭嵌紋病毒 (Cymbidium mosaic virus, CymMV) 及齒舌蘭輪點病毒 (Odontoglossum ringspot virus, ORSV) 鞘蛋白基因之默化載體,轉殖至蝴蝶蘭中,期望獲得抗病之蝴蝶蘭。部分經抗生素篩選存活之蝴蝶蘭擬原球體已再生為植株,葉片經GUS活性組織化學染色分析呈藍色反應。擬轉殖株DNA經聚合酶連鎖反應可合成預期片段909 bp,進一步以南方氏雜交分析,確定得到7棵轉殖蝴蝶蘭。 | zh_TW |
dc.description.abstract | To prolong the longevity of Cattleya by application of biotechnology, inhibition of ethylene signal transduction is used as the strategy. To overexpress negative regulator of ethylene signal transduction MhCTR1 (Musa spp. Constitutive Triple Response 1), driven by CaMV 35S promoter was transformed into Cattleya via Agrobacterium-mediated transformation. Pre-culture condition of explant, concentration of Agrobacterium, time of sonication assistant Agrobacterium-mediated transformation (SAAT), and co-culture days were tested to establish the transformation system of Cattleya. To prove of concept, MhCTR1 was also expressed in Arabidopsis. The T1 transgenic Arabidopsis produced larger flowers with longer flowering period than wild-type. T3 seeds of transgenic Arabidopsis, shown no obvious triple response under ACC treatment, indicated that this strategy inhibit ethylene response. For pre-culture test, the leaves of Cattleya were treated with sonication 15 sec. and cultured in dark for 2 weeks had greater expanded rate. Lower concentration of Agrobacterium used in transformation resulted higher regeneration rate of shoot and lower relapse rate of Agrobacterium. On the other hand, to achieve virus-resistant transgenic Phalaenopsis, the construct express hairpin small RNA of both Cymbidium mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV) coat protein genes based on RNA interference (RNAi) technology was transformed into Phalaenopsis previously. After transgenic plantlet regenerated from antibiotic-resistant Phalaenopsis PLBs had been confirmed by GUS staining and polymerase chain reaction in which expected 909 bp fragment was synthesized. Moreover, 7 transgenic Phalaenopsis have been confirmed by Southern hybridization analysis. | en |
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dc.description.tableofcontents | 壹、 前言 1 貳、 前人研究 2 一、 嘉德麗雅蘭 2 二、 乙烯與更年性花卉 2 三、 乙烯之訊息傳導 2 四、 CTR1基因之相關研究 3 五、 利用基因轉殖降低花辦對乙烯之敏感度 4 六、 利用基因轉殖延緩乙烯造成之花辦老花 5 七、 利用RNA干擾技術提高植物對病毒之抗性 5 (一) 轉殖病毒反義基因以提高植物對病毒之抗性 6 (二) 過量表現順義基因以提高植物對病毒之抗性 6 (三) 同時表現順反義基因以提高植物對病毒之抗性 7 八、 蕙蘭嵌紋病毒 8 九、 齒舌蘭輪點病毒 8 十、 複莖蘭類癒傷組織之誘導 9 參、 材料方法 10 一、 試驗材料 10 (一) 質體 10 (二) 試驗菌種 10 (三) 植物材料 10 1. 阿拉伯芥 (Arabidopsis thaliana ecotype Col-0) 10 2. 嘉德麗雅蘭 10 3. 蝴蝶蘭 10 二、 試驗方法 11 (一) 阿拉伯芥花序轉殖 11 1. 植株栽培 11 2. 農桿菌之製備 11 3. 阿拉伯芥之農桿菌轉殖 11 4. 阿拉伯芥轉殖株之抗生素篩選 12 (二) 阿拉伯芥轉殖株三相反應試驗 12 (三) 嘉德麗雅蘭葉片及擬原球體轉殖 12 1. 嘉德麗雅蘭葉片轉殖流程 12 (1) 預培養前的超音波震盪處理 13 (2) 感染時之菌液濃度測試 13 (3) 最適超音波震盪輔助農桿菌媒介法之測試 13 (4) 最適共培養天數之測試 14 2. 嘉德麗雅蘭葉片ACC誘導老化試驗 14 3. 嘉德麗雅蘭擬原球體轉殖 14 4. 嘉德麗雅蘭原球體誘導癒傷組織 14 (四) 擬轉殖株分析 15 1. GUS活性組織化學染色法 15 2. 植物基因組DNA之抽取 15 3. 聚合酶連鎖反應(polymerase chain reaction, PCR)分析 16 4. 核酸探針之製備和同位素標定 16 5. 南方氏雜交分析 17 肆、 結果 19 一、 35S::MhCTR1之阿拉伯芥基因轉殖 19 (一) 35S::MhCTR1阿拉伯芥轉殖株T1之分子驗證 19 (二) 35S::MhCTR1阿拉伯芥轉殖株T1之外表形態 19 (三) 35S::MhCTR1阿拉伯芥轉殖株T2之抗生素抗性分析 20 (四) 35S::MhCTR1轉殖野生型阿拉伯芥之三相反應 20 二、 嘉德麗雅蘭之基因轉殖 20 (一) 預培養前超音波震盪處理對嘉德麗雅蘭葉片膨大的影響 20 (二) 農桿菌感染時縱切嘉德麗雅蘭葉基部對培植體之影響 21 (三) 超音波震盪輔助農桿菌基因轉殖中菌液濃度及共培養天數對嘉德麗雅蘭芽體再生率之影響 21 (四) 菌液濃度及超音波震盪輔助農桿菌轉殖法對嘉德麗雅蘭芽體再生率之影響 22 (五) ACC處理對嘉德麗雅蘭葉片褐化之影響 23 (六) 2,4-D對誘導嘉德麗雅蘭癒傷組織之影響 23 三、 蝴蝶蘭pGcET-CyORi擬轉殖株之分子驗證 24 伍、 討論 51 一、 阿拉伯芥 51 (一) 抑制乙烯反應對種子發芽之影響 51 (二) 35S::MhCTR1阿拉伯芥轉殖株之外表形態 52 (三) 35S::MhCTR1阿拉伯芥轉殖株於高溫逆境下之變異 53 二、 嘉德麗雅蘭轉殖系統之建立 53 (一) 預培養 53 (二) 感染時切割培植體 54 (三) 菌液濃度 55 (四) 超音波震盪輔助農桿菌轉殖法 56 (五) 共培養 57 (六) 嘉德麗雅蘭葉片轉殖後之ACC處理 57 陸、 結語 59 柒、 參考文獻 63 | |
dc.language.iso | zh-TW | |
dc.title | 延緩花瓣老化及抗病毒蘭花基因轉殖之研究 | zh_TW |
dc.title | Studies on Delay in Petal Senescence and Virus Resistance in Transgenic Orchids | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 黃鵬林(Pung-Ling Huang) | |
dc.contributor.oralexamcommittee | 劉祖惠(Tsu-Hwei Liu),何錦玟(Chin-Wen Ho) | |
dc.subject.keyword | 嘉德麗雅蘭,蝴蝶蘭,阿拉伯芥,CTR1,核糖核酸干擾,蘭花基因轉殖, | zh_TW |
dc.subject.keyword | Cattleya,Phalaenopsis,Arabidopsis,CTR1,RNA interference,orchid transforamtion, | en |
dc.relation.page | 71 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-07-30 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
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