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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張雅君(Ya-Chun Chang) | |
dc.contributor.author | Ting-Yi Kuo | en |
dc.contributor.author | 郭庭禕 | zh_TW |
dc.date.accessioned | 2021-06-16T03:39:54Z | - |
dc.date.available | 2024-12-31 | |
dc.date.copyright | 2015-03-02 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-02-16 | |
dc.identifier.citation | 林正忠、郭韋柏、蔡淑芬。2006。台灣紅龍果病害。豐年 56:38-41。
邱禮弘。2009。紅龍果有機栽培技術。有機農業產業發展研討會專輯:133-139。 邱禮弘、徐敏記。2012。紅龍果。臺灣中部地區外銷作物產業專集:45-54。 黃乃芸。2009。紅龍果。健康世界。176-177頁。 楊玉婷、劉依蓁。2014。臺灣紅龍果生產技術改進研討會採訪紀實。植物種苗生技 37:89-92。 廖吉彥、張清安、顏昌瑞、陳昱初、鄧汀欽。2003。感染紅龍果之仙人掌病毒X之鑑定與分佈調查。植病會刊 12:225-234。 蔡志濃、林筑蘋、安寶貞、鄧汀欽、廖吉彥、倪蕙芳、楊宏仁。2013。紅龍果的重要病害及其防治。農業試驗所技術服務96:1-7。 劉命如。2004。引起紅龍果斑駁病徵之Cactus virus X的鑑定與免疫檢測。植病會刊 13:27-34。 劉碧鵑。2010。台灣紅龍果的栽培。農業試驗所特刊。 Alivisatos, P. 2004. The use of nanocrystals in biological detection. Nat. Biotechnol. 22: 47-52. Amelunxen, F. 1958. Die Virus-Eiweispindeln der Kakteen. Darstellung, elektronenmikroskopische und biochemische Analyse des Virus. Protoplasma. 49: 140-178. Bercks, R. 1971. Cactus virus X. CMI/AAB Descriptions of plant viruses. NO. 58. Brigger, I., Dubernet, C., and Couvreur, P. 2002. Nanoparticles in cancer therapy and diagnosis. Adv. Drug Deliv. Rev. 54: 631-651. Brown, M. A., and Semelka, R. C. 2011. MRI: Basic Principles and Applications. John Wiley & Sons. Casper, R., and Brandes, J. 1969. A new cactus virus. J. Gen. Virol. 5: 155-156. Chen, S. Y., Cheng, L. C., Chen, C. W., Lee, P. H., Yu, F., Zhou, W., Liu, R. S., Do, Y. Y., and Huang, P. L. 2013. NIR-assisted orchid virus therapy using urchin bimetallic nanomaterials in phalaenopsis. Adv. Nat. Sci. Nanosci. Nanotechnol. 4: 045006. Chuang, M. F., Ni, H. F., Yang, H. R., Shu, S. L., and Lai, S. Y. 2012. First report of stem canker disease of pitaya (Hylocereus undatus and H. polyrhizus) caused by Neoscytalidium dimidiatum in Taiwan. Plant Dis.: 906-907. Galarneau, A., Min, K. L., Mangos, M. M., and Damha, M. J. 2005. Assay for evaluating ribonuclease H-mediated degradation of RNA-antisense oligonucleotide duplexes. Methods in Molecular Biology 288: 65-80. Gallo, J., Long, N. J., and Aboagye, E. O. 2013. Magnetic nanoparticles as contrast agents in the diagnosis and treatment of cancer. Chem. Soc. Rev. 42: 7816-7833. Giljohann, D. A., Seferos, D. S., Daniel, W. L., Massich, M. D., Patel, P. C., and Mirkin, C. A. 2010. Gold nanoparticles for biology and medicine. Angew. Chem. Int. Ed Engl. 49: 3280-3294. Giraldo, J. P., Landry, M. P., Faltermeier, S. M., McNicholas, T. P., Iverson, N. M., Boghossian, A. A., Reuel, N. F., Hilmer, A. J., Sen, F., Brew, J. A., and Strano, M. S. 2014. Plant nanobionics approach to augment photosynthesis and biochemical sensing. Nat. Mater. 13: 400-408. Kim, B., Han, G., Toley, B. J., Kim, C., Rotello, V. M., and Forbes, N. S. 2010. Tuning payload delivery in tumour cylindroids using gold nanoparticles. Nat. Nanotechnol. 5: 465-472. King, A. M. Q., Adams, M. J., Carstens, E. B., and Lefkowitz, E. J. 2012. Virus Taxonomy. Ninth Report of the International Committee on Taxonomy of Viruses. PP. 912-915. Koenig, R., Pleij, C. W. A., Loss, S., Burgermeister, W., Aust, H., and Schiemann, J. 2004. Molecular characterisation of potexviruses isolated from three different genera in the family Cactaceae. Arch. Virol. 149: 903-914. Lee, C. C. 2014. Analysis and application of cis-acting elements and defective RNA of Pitaya virus X. Master’s thesis, National Taiwan University, Taipei, Taiwan. Lewinski, N., Colvin, V., and Drezek, R. 2008. Cytotoxicity of nanoparticles. Small Weinh. Bergstr. Ger. 4: 26-49. Liou, M. R. 2000. Characterization of Pitaya mosaic virus and analysis of its full-length sequence. Master’s thesis, National Taiwan University, Taipei, Taiwan. Liou, M. R., Chen, Y. R., and Liou, R. F. 2004. Complete nucleotide sequence and genome organization of a Cactus virus X strain from Hylocereus undatus (Cactaceae). Arch. Virol. 149: 1037-1043. Li, Y. S. 2010. Characterization, infectious clone construction and antiserum preparation of Pitaya virus X. Master’s thesis, National Taiwan University, Taipei, Taiwan. Lu, Y. C. 2007. Characterization of a new Cactus virus X isolate from pitaya and the construction of its infectious cDNA clone. Master’s thesis, National Taiwan University, Taipei, Taiwan. Mao, C. H. 2008. Molecular characterization and detection of new Zygocactus virus X and Pitaya virus X from pitaya. Master’s thesis, National Taiwan University, Taipei, Taiwan. Medintz, I. L., Uyeda, H. T., Goldman, E. R., and Mattoussi, H. 2005. Quantum dot bioconjugates for imaging, labelling and sensing. Nat. Mater. 4: 435-446. Merten, S. 2003. A review of Hylocereus production in the United States. J PACD. 5: 98-105. Mizrahi, Y., Nerd, A. and Nobel, P. S. 1997. Cacti as crop. Horticultural reviews 18: 291-391. Palmateer, A. J., Ploetz, R. C., van Santen, E., and Correll, J. C. 2007. First occurrence of anthracnose caused by Colletotrichum gloeosporioides on pitahaya. Plant Dis. 91: 631-631. Peer, D., Karp, J. M., Hong, S., Farokhzad, O. C., Margalit, R., and Langer, R. 2007. Nanocarriers as an emerging platform for cancer therapy. Nat. Nanotechnol. 2: 751-760. Pisanic, T. R., Blackwell, J. D., Shubayev, V. I., Fiñones, R. R., and Jin, S. 2007. Nanotoxicity of iron oxide nanoparticle internalization in growing neurons. Biomaterials 28: 2572-2581. Singh, N., Jenkins, G. J. S., Asadi, R., and Doak, S. H. 2010. Potential toxicity of superparamagnetic iron oxide nanoparticles (SPION). Nano Rev. 1: 10.3402. Takahashi, L. M., Rosa, D. D., Basseto, M. A., de Souza, H. G., and Furtado, E. L. 2008. First report of Colletotrichum gloeosporioides on Hylocereus megalanthus in Brazil. Australas. Plant Dis. Notes. 3: 96-97. Wang, C. L., and Lin, C. C. 2005. Fruit rot of pitaya and stem rot of cacti in Taiwan. Plant Pathol. Bull. 14: 269-274. Wang, Z., Liu, H., Yang, S. H., Wang, T., Liu, C., and Cao, Y. C. 2012. Nanoparticle-based artificial RNA silencing machinery for antiviral therapy. Proc. Natl. Acad. Sci. 109: 12387-12392. Weissleder, R., Stark, D., Engelstad, B., Bacon, B., Compton, C., White, D., Jacobs, P., and Lewis, J. 1989. Superparamagnetic iron oxide: pharmacokinetics and toxicity. Am. J. Roentgenol. 152: 167-173. Xu, H., Wang, Q. L., Wei, G., and Mo, J. G. 2010. The health benefits and research progress of pitaya. Journal of Guangxi Academy of Science. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/54852 | - |
dc.description.abstract | 紅龍果 (dragon fruit; pitaya) 屬於仙人掌科 (Cactaceae)、三角柱屬 (Hylocereus spp.) 的多年生攀緣性肉質植物,由於紅龍果外觀討喜,果實風味佳,且具高營養價值,加上目前台灣積極推廣紅龍果的外銷,使紅龍果成為極具市場經濟潛力的熱帶果樹。台灣目前多以扦插方式進行繁殖,故若作為扦插原的紅龍果母本為帶病毒植株,即會使病毒在田間快速擴散。根據前人研究,Potexvirus屬的仙人掌X病毒 (Cactus virus X, CVX)為台灣第一個被報導可以感染紅龍果的病毒,2006年本實驗室於陽明山觀光果園的紅龍果發現除了CVX外,尚有在台灣未曾報導過的蟹爪蘭X病毒 (Zygocactus virus X, ZyVX),以及一種新的potexvirus病毒,命名為紅龍果X病毒 (Pitaya virus X, PiVX)。由田間調查顯示這三種病毒常感染紅龍果,而Potexvirus屬病毒也是目前唯一被報導會感染紅龍果的病毒屬。本研究利用羧基 (-COOH) 修飾之四氧化三鐵 (Fe3O4) 磁性奈米粒子,開發出一套針對紅龍果中三種potexvirus病毒之快速檢測方法,命名為magnetic nanoparticle-capture reverse transcription-polymerase chain reaction (MNC RT-PCR),以利往後病毒檢測。也藉由此檢測方法對其他同為仙人掌科之十種常見觀賞仙人掌:蟹爪蘭、白烏帽子、金烏帽子、紅牡丹、金鯱、兜丸、劍戀玉、岩石獅子、六角柱仙人掌與神仙堡進行檢測,以及對多處台灣紅龍果產區,包含宜蘭、彰化、台東與屏東之紅龍果進行田間調查,分析各地區紅龍果植株的病毒感染率與分布情形。根據目前調查結果顯示,發現蟹爪蘭遭受CVX以及ZyVX感染,將這兩種病毒基因體片段選殖並定序後,於NCBI上進行BLASTn與BLASTx分析。結果發現感染蟹爪蘭的ZyVX與紅龍果中ZyVX-P39在核酸序列以及RdRp與TGB1-3的胺基酸序列相同度皆達95%以上。而CVX的部分RdRp序列與前人在關西果園中發現之CVX-Hu序列相同度達95%,但CP胺基酸序列卻是與韓國之CVX-Jeju3分離株較相似,序列相同度高達99%。本次調查證明此三種potexvirus病毒也存在於紅龍果外的其他觀賞仙人掌上。而田間調查部分,由檢測結果可知除台中農試所外,田間已不存在健康紅龍果植株,推斷是由於使用帶病毒扦插枝條,加上長期使用未經消毒之工具進行連續採收與修枝,導致病毒以機械傳播方式擴散至全園區。故本研究也擬開發一套奈米粒子療法,同樣利用修飾 –COOH 之Fe3O4磁性奈米粒子作為主體,以具有專一辨識性的單股寡核苷酸片段與核糖核酸酶H (RNase H) 進行修飾,藉由對三種potexvirus RNA的專一性雜合,使磁性奈米粒子表面上鄰近之RNase H能夠截切目標病毒RNA受雜合之複製相關重要區域,以此降低受感染紅龍果中的potexviruses病毒複製能力,達到去除病毒的目的。奈米療法部分已完成辨識性寡核苷酸片段之選定與粒子修飾條件測試,將合成之粒子命名為Nanozyme,並已進行初步生體外去病毒RNA試驗。目前所使用之Nanozyme確實具有專一性辨識能力,但在裁切病毒RNA能力上效率不顯著,仍需進行實驗條件的測試與改良。 | zh_TW |
dc.description.abstract | Pitaya, also called dragon fruit, which is a kind of perennial climbing succulent plant belongs to the genus Hylocereus in the family of Cactaceae. Because of its charming, delicious fruit with high nutrition and the export policy of government, pitaya has become an important tropical fruit crop in Taiwan. Pitaya is mainly propagated by cutting, so if mother plant is infected with viruses, disease can be spread easily. According to the previous studies, Cactus virus X (CVX), belonging to the genus Potexvirus, is the first pitaya-infecting virus reported in Taiwan. The other two potexviruses, Zygocactus virus X (ZyVX), first reported in Taiwan, and Pitaya virus X (PiVX), a new species of potexvirus, were first identified by our laboratory in the Yanmingshan orchard in 2006. The previous field survey indicated that these three potexviruses infect pitaya frequently, and so far Potexvirus genus is the only one virus genus reported to infect pitaya plants. In this study, a rapid detection method, named magnetic nanoparticle-capture reverse transcription-polymerase chain reaction (MNC RT-PCR), using Fe3O4 magnetic nanoparticles modified with –COOH for detecting the pitaya-infecting potexviruses was developed. Ten common ornamental Cactaceae plants, including Opuntia microdasys v. albispina Fobe., Opuntia microdasys, Gymnocalycium mihanovichii var. friedrichii, Echinocactus grusonii, Astrophytum asterias Lem., Echinofossulocactus kellerianus Krainz, Cereus peruvianus f. monstrosus, Cereus peruvianus Mill. and Cereus 'Fairy Castle', were tested for virus infection by MNC RT-PCR. Pitaya plants from several production areas in Taiwan, including Ilan, Changhua, Taitung, Pingtung and Taichung, were also detected by MNC RT-PCR to investigate the infection rate and distribution of potexviruses. The results of field survey revealed that zygocactus can be infected by CVX and ZyVX, and these three potexviruses can infect other Cactaceae plants. After cloning and sequencing the fragments of the two potexviruses from zygocactus and BLAST in NCBI database, it shows that the nucleotide sequence identity and amino acid sequence identities of RdRp and TGB1-3 ORF are above 95% between zygocactus ZyVX and pitaya ZyVX-P39. The nucleotide sequence identity of partial RdRp is 95% between zygocactus CVX and pitaya CVX-Hu, whereas CP amino acid sequence of zygocactus CVX is similar to CVX-Jeju3 from Korea with 99% identity. The field survey of pitaya shows that healthy pitaya plants could not discovered in the field except pitaya samples collected from Taichung Agricultural Research Institution. It may be that farmers use potexvirus-infected cuttings as propagules, and mechanical inoculation through contaminated tools cutting stems or harvesting fruits successively without disinfection. Due to the high infection rate of potexvirus in pitaya, the other target in this thesis is to develop a magnetic nanoparticle therapy by using Fe3O4–COOH modified with specific single strand oligonucleotides and Ribonuclease H (RNase H). When the target potexviruses are hybridized with specific oligonucleotides, the near RNase H can degrade hybridized viral RNA, to reduce the replication ability of target potexviruses. So far the sequences of specific identification oligonucleotides were determined, and the modification condition of Fe3O4–COOH, named the modified nanoparticle Nanozyme, was tested. The preliminary efficiency test in vitro indicated Nanozyme shows its specificity to recognize target potexviruses, but its degradation efficiency to target fragment is not obvious. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T03:39:54Z (GMT). No. of bitstreams: 1 ntu-104-R01645003-1.pdf: 4984906 bytes, checksum: 69672036688481aff983dd0e12f3e6c6 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 目錄
中文摘要 V Abstract VII 壹、前言 1 一、紅龍果 1 (一) 紅龍果特性 1 (二) 生長條件與生育特性 1 (三) 營養價值 1 (四) 經濟重要性 2 (五)常見紅龍果病害 2 二、馬鈴薯X病毒屬 (Potexvirus) 3 (一) Potexvirus特性 3 (二) 感染紅龍果之potexviruses 3 三、磁性奈米粒子 4 (一) 奈米粒子定義與應用 4 (二) 氧化鐵系列磁性奈米粒子特性 5 (三) 奈米粒子於植物上之研究 5 四、研究動機 6 貳、材料及方法 7 一、以間接酵素連結抗體免疫吸附反應 (Indirect-ELISA) 檢測紅龍果枝條 7 二、以組織轉印免疫法分析病毒在植物組織中之分布情形 7 三、研發紅龍果potexvirus病毒之快速簡易檢測方法 8 (一) 不同研磨緩衝液之種類與體積對MNC RT-PCR檢測效果之測試 8 (二) 液態氮使用與否對MNC RT-PCR檢測效果之測試 9 (三) 測試磁性奈米粒子之合適使用量 9 四、以新研發之MNC RT-PCR檢測仙人掌科植物樣品 9 (一) 以MNC RT-PCR檢測不同仙人掌科作物 9 (二) 以MNC RT-PCR檢測田間紅龍果植株 10 五、選殖並分析蟹爪蘭植物中之CVX與ZYVX 10 (一) 植物全RNA 之純化 10 (二) 反轉錄聚合酶連鎖反應 11 (三) 回收聚合酶連鎖反應產物之DNA片段 11 (四) 目標DNA片段之黏合與轉型反應 12 (五) 轉型株之篩選 13 (六) 質體之小量製備 13 六、磁性奈米粒子去病毒療法之研發 14 (一) 設計並合成辨識目標病毒之寡核苷酸片段 14 (二) 寡核苷酸片段與目標病毒之專一性辨識能力測試 14 1. 製備病毒缺失性RNA之生體外轉錄體 14 2. RNase H assay 15 (三) 將寡核苷酸片段鍵結於磁性奈米粒子上 15 (四) 以標定螢光染劑之探針觀察磁性奈米粒子修飾情形 16 (五) Nanozyme之合成 16 (六) Nanozyme於生體外截切效率與專一性之測試 17 (七) 磁性奈米粒子對植物之毒性測試 17 參、結果 18 一、以間接酵素連結免疫反應分析健康及受potexvirus病毒感染之紅龍果枝條 18 二、以組織轉印免疫法分析病毒在紅龍果肉質莖中分布情形 18 三、研發紅龍果potexvirus病毒之快速簡易檢測方法 18 (一) 不同研磨緩衝液之種類與體積對MNC RT-PCR檢測結果之影響 18 (二) 使用液態氮與否對MNC RT-PCR檢測效果之測試 19 (三) 測試磁性奈米粒子合適使用量 19 四、以新研發之MNC RT-PCR檢測市售仙人掌科植物與田間紅龍果植株 20 (一) 以MNC RT-PCR檢測不同仙人掌科植物 20 (二) 以MNC RT-PCR檢測田間紅龍果potexvirus之分布 21 五、選殖並分析蟹爪蘭植物中之CVX與ZYVX 22 六、磁性奈米粒子去病毒療法之研發 24 (一) 選擇並合成可辨識目標病毒之寡核苷酸片段 24 (二) 寡核苷酸片段與目標病毒之專一性辨識能力測試 24 (三) 磁性奈米粒子修飾寡核苷酸片段之效率 25 (四) 以標定螢光染劑之探針觀察磁性奈米粒子之修飾情形 26 (五) Nanozyme之合成 26 (六) Nanozyme於生體外截切病毒RNA之效率與專一性 27 (七) 磁性奈米粒子對植物之毒性測試 27 肆、討論 29 一、以免疫法分析紅龍果枝條 29 二、紅龍果potexvirus病毒快速簡易檢測方法之研發 29 三、不同仙人掌科植物檢測 32 四、紅龍果田間調查 32 五、分析蟹爪蘭植物中之CVX與ZYVX 35 六、磁性奈米粒子去病毒療法之研發 35 (一) 辨識目標病毒的寡核苷酸片段之選擇與合成 36 (二) Nanozyme之合成條件 36 (三) Nanozyme之效率與專一性 37 (四) 磁性奈米粒子之植物毒性分析 38 七、結語 39 伍、參考文獻 40 陸、附表 45 柒、附圖 50 附錄 77 | |
dc.language.iso | zh-TW | |
dc.title | 紅龍果potexviruses病毒快速檢測方法與磁性奈米粒子療法之開發及應用 | zh_TW |
dc.title | Rapid detection method and magnetic nanoparticle therapy for pitaya-infecting potexviruses | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 洪挺軒,陳煜焜(Yuh-Kun Chen) | |
dc.subject.keyword | 仙人掌X病毒,磁性奈米粒子,紅龍果,紅龍果X病毒,馬鈴薯X病毒屬,蟹爪蘭X病毒, | zh_TW |
dc.subject.keyword | Cactus virus X (CVX),magnetic nanoparticle (MNP),pitaya,Pitaya virus X (PiVX),Potexvirus,Zygocactus virus X (ZyVX), | en |
dc.relation.page | 84 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-02-16 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 植物醫學碩士學位學程 | zh_TW |
顯示於系所單位: | 植物醫學碩士學位學程 |
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