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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 洪挺軒(Ting-Hsuan Hung) | |
| dc.contributor.author | Yu-Chen Liu | en |
| dc.contributor.author | 劉祐辰 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:35:49Z | - |
| dc.date.available | 2023-10-26 | |
| dc.date.copyright | 2021-11-02 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-26 | |
| dc.identifier.citation | 行政院農業委員會。2018年。百香果產業概況。農業知識入口網，百香果主題館。https://kmweb.coa.gov.tw/subject/index.php?id=161 （上網日期：2021.8.19） 吳孟玲。1994。單元抗體之製備與應用於香蕉嵌紋病原胡瓜嵌紋病毒之系統與感染生態研究。國立臺灣大學植物病蟲害學研究所博士論文。176頁。 吳柏辰。2019。建立香蕉病毒之四合一反轉錄聚合酶連鎖反應法檢測及調查臺灣香蕉之病毒感染情形。國立臺灣大學植物病理與微生物學研究所碩士論文。87頁。 李文立、Chinnapan Thanarut、徐智政、李國基、江芬蘭、陳金枝。2020。夜香花嵌紋病毒百香果分離株分子特性及廣效性分子檢測技術之開發應用。臺灣農業研究 69(1):46-64頁。 林盈達。1982。百香果雜交F1品種之育成。中國農藝 28:190頁。 張清安、王惠亮、周定芸、陳玉蓉。1981。百香果毒素病之調查與鑑定。植物保護學會刊 23(4):267頁。 張清安、林盈達。1992。無病毒百香果苗栽植後之再感染生態及其防治病毒病效果評估。植物病理學會刊 1:140-146頁。 陳金枝、江芬蘭、黃美容。2017a。促進百香果病毒East Asian passiflora virus檢出效果之植株取樣部位與病毒多元抗體應用方式。106年植保年會論文宣讀摘要 59(4): 41頁。 陳金枝、江芬蘭、鄭櫻慧、鄧汀欽。2015。可檢測多種百香果potyviruses之多元抗體製備與應用。104年植保年會論文宣讀摘要 35頁。 陳金枝、鄭櫻慧、鄧汀欽、江芬蘭。2017b。百香果病毒診斷與檢測試劑產業應用。農業試驗所技術服務季刊 112:9-14頁。 馮雅智。2006。香蕉新病毒－香蕉苞葉嵌紋病毒之鑑定及分佈調查。國立臺灣大學植物病蟲害學研究所博士論文。88頁。 黃莉真。2014。可同時檢測多種百香果病毒之多目標型增幅及晶片檢測系統朝陽科技大學應用化學系生化科技碩士班碩士論文。101頁 蔡一慈。2017。夜香花嵌紋病毒在臺灣百香果上的分離、鑑定與檢測技術的發展。國立中興大學國際農學碩士學位學程碩士論文。94頁。 鄭櫻慧、陳金枝、鄧汀欽。2015。百香果雙生病毒發生現況。臺灣新浮現之重要作物病害及其防治研討會專刊184:99-106頁。 Abdullah, N., Ismail, I., Pillai, V., Abdullah, R., and Sharifudin, A. S. 2009. Nucleotide sequence of the coat protein gene of the Malaysian passiflora virus and its 3′ non-coding region. Am. J. Appl. Sci. 6(9):1633-1636. Adams, M. J., Antoniw, J. F., and Fauquet, C. M. 2005. Molecular criteria for genus and species discrimination within the family Potyviridae. Arch. Virol. 150(3):459-479. Brand, R. J., and Wechmar, M. B. von. 1993. Characterization of two viruses implicated in the woodiness disease complex of South African passionfruit: Cucumber mosaic virus and a new potyvirus. J. South. Afr. Soc. Hortic. Sci. 3(1):28-33. Brown, J. K., Bird, J., and Fletcher, D. C. 1993. First report of passiflora leaf mottle disease caused by a whitefly-transmitted geminivirus in Puerto Rico. Plant Dis. 77(42):1264 Chang, C. A. 1992. Characterization and comparison of passionfruit mottle virus, a newly recognized potyvirus, with Passionfruit woodiness virus. Phytopathology 82(11):1358-1363. Chang, C. A., Chen, C. C., Deng, T. C., and Zettler, F. W. 1996. Characterization of passionfruit crinkle potyvirus-a newly found virus infecting passionfruit. Plant Prot. Bull. 38(4):339-354. Chang, C. A., Chen, C. M., and Wang, H. L. 1987. Identification of a newly recognized potyvirus causing passionfruit mottling. Plant Prot. Bull. 29:445. Chang, C. A., Huang, L. C., Chen, Y. C., Lin, Y. Y., Lu, H. C., and Lin, Y. D. 2017. A review of the research and control of passionfruit virus diseases in Taiwan. J. Plant Med. 59(3):1-12. Chen, S., Yu, N., Yang, S., Zhong, B., and Lan, H. 2018. Identification of telosma mosaic virus infection in Passiflora edulis and its impact on phytochemical contents. Virol. J. 15(1):1-8. Cheng, Y. H., Deng, T. C., Chen, C. C., Chiang, C. H., and Chang, C. A. 2014. First Report of Euphorbia leaf curl virus and Papaya leaf curl Guangdong virus on passion fruit in Taiwan. Plant Dis. 98(12):1746. Chiemsombat, P., Prammanee, S., and Pipattanawong, N. 2014. Occurence of telosma mosaic virus causing passion fruit sever mosaic disease in Thailand and immunostrip test for rapid virus detection. Crop Prot. 63:41-47. Chong, Y. H., Cheng, Y. H., Cheng, H. W., Huang, Y. C., and Yeh, S. D. 2018. The virus causing passionfruit woodiness disease in Taiwan is reclassified as East Asian passiflora virus. J. Gen. Plant Pathol. 84(3):208-220 Dougherty, W. G., Cary, S. M., and Parks, T. D. 1989. Molecular genetic analysis of a plant virus polyprotein cleavage site: a model. Virol. 171(2):356-364. Ferreira, S. S., Barros, D. R., de Almeida, M. R., and M. Zerbini, F. M. 2010. Characterization of Passionfruit severe leaf distortion virus, a novel begomovirus infecting passionfruit in Brazil, reveals a close relationship with tomato-infecting begomoviruses. Plant Pathol. 59(2):221-230. Fukumoto, T., Nakamura, M., Rikitake, M., and Iwai, H. 2012. Molecular characterization and specific detection of two genetically distinguishable strains of East Asian Passiflora virus (EAPV) and their distribution in southern Japan. Virus genes 44(1):141-148. Gibbs, A. J., Trueman, J. W. H., and Gibbs, M. J. 2008. The been common mosaic virus lineage of potyviruses: where did it arise and when? Arch. Virol. 153(12):2177-2187. Ha, C., Coombs, S., Revill, P. A., Harding, R. M., Vu, M., and Dale, J. L. 2008. Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses. Arch. Virol. 153(1):25-36. Hari, V., Siegel, A., Rozek, C., and Timberlake, W.E. 1979. The RNA of tobacco etch virus contains poly (A). Virol. 92(2):568-571. Hung, T. H., Wu, N. M., and Su, H. J. 1999. Development of a rapid method for the diagnosis of citrus greening disease using the polymerase chain reaction. J. Phytopathology 147(10):599-604. Iwai, H., Terahara, R., Yamashita, Y., Ueda, S., and Nakamura, M. 2006a. Complete nucleotide sequence of the genomic RNA of an Amami-O-shima strain of East Asian Passiflora potyvirus. Arch. Virol. 151(7):1457-1460. Iwai, H., Yamashita, Y., Nishi, N., and Nakamura, M. 2006b. The potyvirus associated with the dappled fruit of Passiflora edulis in Kagoshima prefecture, Japan is the third strain of the proposed new species East Asian Passiflora virus phylogenetically distinguished from strains of Passion fruit woodiness virus. Arch. Virol. 151(4):811-818. Jacob, T., and Usha, R. 2002. Expression of Cardamom mosaic virus coat protein in Escherichia coli and its assembly into filamentous aggregates. Virus Res. 86(1-2):133-141. Jagadish, M. N., Ward, C. W., Gough, K. H., Tulloch, P. A., Whittaker, L. A., and Shukla, D. D. 1991. Expression of potyvirus coat protein in Escherichia coli and yeast and its assembly into virus-like particles. J. Gen. Virol. 72(7):1543-1550. Jan, F. J. and Yeh, S. D. 1995. Purification, in situ localization, and comparative serological properties of passionfruit woodiness virus-encoded amorphous inclusion protein and two other virus proteins. Phytopathology 85(1):64-71. Joseph, J., and Savithri, H. S. 1999. Determination of 3′-terminal nucleotide sequence of pepper\break vein banding virus RNA and expression of its coat protein in Escherichia coli. Arch. Virol. 144(9):1679-1687. Lee, S. C. and Chang, Y. C. 2006. Multiplex RT-PCR detection of two orchid viruses with an internal control of plant nad5 mRNA. Plant Pathology Bulletin 15(3):187-196. Lin, N. S., Chen, Y. K., and Hsu, Y. H. 1989. Immunological detection of passionfruit woodiness virus. Bot. Bull. Acad. Sinica 30:31-37. Maciel, S. D. C., Nakano, D. H., Rezende, J. A. M., and Vieira, M. L. C. 2009. Screening of Passiflora species for reaction to Cowpea aphid-borne mosaic virus reveals an immune wild species. Sci. Agric. 66(3):414-418. McKern, N. M., Strike, P. M., Barnett, O. W., Dijkstra, J., Shukla, D. D., and Ward, C. W. 1994. Cowpea aphid borne mosaic virus-Morocco and South African Passiflora virus are strains of the same potyvirus. Arch. Virol. 136(1):207-217. McKnight, T. 1953. The woodiness disease of the passion vine (Passiflora edulis Sims). Queensland J. of Agri. Soc. 10(1):4-35. Nascimento, A. V. S., Santana, E. N., Braz, A. S. K., Alfenas, P. F., Pio-Ribeiro, G., Andrade, G. P., Carvalho, M. G., and Zerbini, F. M. 2006. Cowpea aphid-borne mosaic virus (CABMV) is widespread in passionfruit in Brazil and causes passionfruit woodiness disease. Arch. Virol. 151(9):1797-1809. Novaes, Q. S., Freitas‐Astua, J., Yuki, V. A., Kitajima, E. W., Camargo, L. E. A., and Rezende, J. A. M. 2003. Partial characterization of a bipartite begomovirus infecting yellow passion flower in Brazil. Plant Pathol. 52(5):648-654. Omatsu, N., Iwai, H., Setokuchi, O., and Arai, K. 2004. Immigrating aphid species and their importance as vectors of Passionfruit woodiness virus in the fields of Amami Oshima island, Japan. Mem. Fac. Agric. Kagoshima. Univ. 39:1-5. Peng, Y. H., Kadoury, D., Gal-On, A., Huet, H., Wang, Y., and Raccah, B. 1998. Mutations in the HC-Pro gene of zucchini yellow mosaic potyvirus: effects on aphid transmission and binding to purified virions. J. Gen. Virol. 79(4):897-904. Prione, T. P., and Blanc, S. 1996. Helper-dependent vector transmission of plant viruses. Annu. Rev. Phytopathology 34(1):227-247. Riechmann, J. L., Lain, S., and Garcia, J. A. 1992. Highlights and prospects of potyvirus molecular biology. J. Gen. Virol. 73(1):1-16. Shukla, D. D., and Ward, C. W. 1989. Identification and classification of potyviruses on the basis of coat protein sequence data and serology. Arch. Virol. 106(3):171-200. Shukla, D. D., Strike, P. M., Tracy, S. L., Gough, K. H., and Ward, C. W. 1988. The N and C termini of the coat proteins of potyviruses are surface-located and the N terminus contains the major virus-specific epitopes. J. Gen. Virol. 69(7):1497-1508. Siaw, M. F., Shahabuddin, M., Ballard, S., Shaw, J. G. and Rhoads, R. E. 1985. Identification of a protein covalently linked to the 5’ terminus of tobacco vein mottling virus RNA. Virol. 142(1):134-143. Taylor, R. H., and Greber, R. S. 1973. Passion fruit woodiness virus. CMI/AAB. Description of Plant Viruses 122(4) Tsai, W. S., Shih, S. L., Kenyon, L., Green, S. K., and Jan, F. J. 2011. Temporal distribution and pathogenicity of the predominant tomato‐infecting begomoviruses in Taiwan. Plant Pathol. 60(4):787-799. Wylie, S. J., Adamns, M., Chalam, C., Kreuze, J., Lopez-Moya, J. J., Ohshima, K., Praveen, S., Rabenstein, F., Stenger, D., Wang, A., and Murilo Zerbini, F. 2017. ICTV virus taxonomy profile: Potyviridae. J. Gen. Virol. 98(3):352-354. Xie, L., Zhang, X., Zheng, S., Zhang, L., and Li, T. 2017. Molecular identification and specific detection of telosma mosaic virus infecting passion fruit. Scientia Agricultura Sinica 50(24):4725-4734. Yang, K., Yan, H., Song, L., Jin, P., Miao, W., and Cui, H. 2018. Analysis of the complete genome sequence of a potyvirus from passion fruit suggests its taxonomic classification as a member of a new species. Arch. Virol. 163(9):2583-2586. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82099 | - |
| dc.description.abstract | "百香果種苗繁殖及銷售是整個百香果產業的重要經濟利基，臺灣本土每年種苗需求量約為90萬株，其餘種苗多外銷往東南亞，年出口量達800萬株。為了防止病害隨著種苗外銷而蔓延，無病毒健康種苗的生產及病毒檢測極為關鍵。病毒危害對百香果的栽培影響巨大，其中又以東亞百香果病毒 (east asian passiflora virus, EAPV) 造成的百香果木質化病最為嚴重，受感染的植株會有生長勢低落、葉片嵌紋、果實木質化、畸型化、果腔縮小、果汁風味變差⋯⋯等病徵。相較於RT-PCR (reverse transcription-polymerase chain reaction)，利用免疫學原理之酵素連結免疫吸附法 (enzyme-linked immunosorbent assay, ELISA) 為一成本較低、效率較高的病毒檢測方法。因此，本研究針對EAPV製備高敏感度及高專一性之單株抗體，以應用於病毒免疫檢測。將EAPV之鞘蛋白基因選殖至表現載體pET-28a(+)，並以大腸桿菌系統 (Escherichia coli BL21 (DE3)) 表現重組鞘蛋白作為免疫抗原，注射小鼠產生免疫反應，經細胞融合技術，篩選獲得一單元抗體細胞株mAb7H8G9。經間接酵素連結免疫吸附法 (indirect-ELISA) 分析，純化後之抗體對EAPV病葉粗萃取液的力價可達16384倍，並且能夠檢測到稀釋640倍的病葉粗萃取液。藉由田間百香果病毒調查，檢視所設計之ELISA檢測方法的效力，顯示mAb7H8G9對臺灣主要栽培品種台農一號的檢測正確率高。另外，也針對EAPV之AO strain與IB strain都具有的保守區域以及歧異度較大的區域，分別設計出EAPV廣效性引子對，以及兩strain各自的專一性引子對，並首次建立EAPV RT-qPCR (real-time RT-PCR)。本研究研發不同面向的檢測方法，可提供未來學術研究與百香果防檢疫之應用。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T05:35:49Z (GMT). No. of bitstreams: 1 U0001-2410202115224200.pdf: 15961866 bytes, checksum: dd593d74c51db7a75443a572a3b5d31f (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 誌謝 i 摘要 ii Abstract iii 目錄 v 表目錄 viii 圖目錄 ix 壹、前言 1 貳、前人研究 3 一、 百香果之簡介 3 二、 百香果病毒種類 4 (一) Potyvirus 4 (二) 臺灣百香果之potyvirus 5 (三) East asian passiflora virus 6 (四) Cowpea aphid-borne mosaic virus 6 (五) Telosma mosaic virus 7 (六) 臺灣百香果之begomovirus 7 三、 東亞百香果病毒檢測方式 8 參、材料與方法 9 一、 病毒的來源及保存 9 二、 分子特性分析 9 (一) 植物總核酸萃取法 9 (二) 專一性引子對的設計 10 (三) 反轉錄聚合酵素連鎖反應 11 (四) 基因選殖 11 (五) 即時反轉錄聚合酵素連鎖反應 12 三、 重組病毒鞘蛋白之表現 13 (一) 表現載體的建構 13 (二) 重組鞘蛋白之誘導表現及純化 14 (三) 蛋白質電泳分析 15 (四) 西方轉漬免疫法 16 四、 單元抗體之製備 16 (一) 動物免疫注射 16 (二) 細胞融合 17 (三) 細胞篩選 18 (四) 細胞單株化 18 (五) 細胞冷凍保存與解凍 19 (六) 單元抗體之生產 19 (七) 單元抗體亞型之測定與抗體純化 20 五、 單元抗體之特性分析 20 (一) 以間接酵素連結免疫吸附法測定抗體力價 20 (二) 以間接酵素連結免疫吸附法測定不同EAPV病毒萃取溶液 20 (三) 以間接酵素連結免疫吸附法測定抗體之可偵測稀釋臨界點 21 (四) 單元抗體與酵素alkaline phosphatase連結 21 (五) 以三明治酵素連結免疫吸附法測試酵素連結之單元抗體 22 六、 單元抗體之應用 22 (一) 東亞百香果病毒間接酵素連結免疫吸附法 22 (二) 東亞百香果病毒三明治酵素連結免疫吸附法 23 (三) 田間百香果病毒病害調查 23 肆、結果 24 一、 EAPV重組鞘蛋白之表現 24 二、 單元抗體株之獲得 24 三、 mAb7H8G9特性分析 25 (一) mAb7H8G9之亞型 25 (二) mAb7H8G9之專一性 25 (三) mAb7H8G9細胞培養上層液之力價 26 (四) mAb7H8G9純化後抗體之力價 26 四、 EAPV ELISA條件測試 26 (一) 不同萃取緩衝溶液對EAPV病毒萃取效果之比較 26 (二) mAb7H8G9對EAPV罹病葉片萃取液之可偵測稀釋臨界點 27 (三) 以DAS-ELISA測試酵素連結之mAb7H8G9 27 五、 mAb7H8G9對感染不同病毒之百香果葉片專一性測試 27 六、 Two step RT-PCR專一性引子對測試 28 七、 RT-qPCR標準曲線之建立及敏感度測試 28 八、 百香果病毒病害田間調查 29 伍、討論 30 陸、參考文獻 36 柒、表 43 捌、圖 54 玖、附錄 69 | |
| dc.language.iso | zh-TW | |
| dc.subject | 重組鞘蛋白 | zh_TW |
| dc.subject | 東亞百香果病毒 | zh_TW |
| dc.subject | 單株抗體 | zh_TW |
| dc.subject | 酵素連結免疫吸附法 | zh_TW |
| dc.subject | recombinant coat protein | en |
| dc.subject | east asian passiflora virus | en |
| dc.subject | monoclonal antibody | en |
| dc.subject | ELISA | en |
| dc.title | 東亞百香果病毒單株抗體與分子檢測方法之研發及應用 | zh_TW |
| dc.title | Development and application of the monoclonal antibody and molecular detections for east asian passiflora virus | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳煜焜(Hsin-Tsai Liu),葉信宏(Chih-Yang Tseng),張雅君 | |
| dc.subject.keyword | 東亞百香果病毒,單株抗體,酵素連結免疫吸附法,重組鞘蛋白, | zh_TW |
| dc.subject.keyword | east asian passiflora virus,monoclonal antibody,ELISA,recombinant coat protein, | en |
| dc.relation.page | 75 | |
| dc.identifier.doi | 10.6342/NTU202104087 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-10-27 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
| dc.date.embargo-lift | 2023-10-26 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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