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標題: 洋桔梗壞疽病毒彩色海芋分離株之鑑定與特性分析
Identification and characterization of a calla lily-infecting isolate of Lisianthus necrosis virus
作者: 張怡珊
Chang, Yi-Shan
關鍵字: Lisianthus necrosis virus
Zantedeschia spp.
出版社: 植物病理學系所
引用: 何楊修. 1993. 海芋 (Zantedeschia spp.) 之種類及栽培習性 (一). 種苗科技專訊5: 4-5. 何陽修、劉明宗、陳駿季、沈再發. 2000. 海芋栽培技術手冊. 農委會種苗改良殖場發行. 台中. 陳金枝、張清安、林玫珠、方懷聖. 2000. 胡瓜嵌紋病毒感染引起的港口馬鈴紋病之鑑定. 植物病理學會刊 9: 29-34. 陳慶忠、柯文華、陳煜焜. 1998. 洋桔梗上胡瓜嵌紋病毒之鑑定及傳播試驗.中區農改場研究彙報60: 1-18. 陳駿季、楊佐琦、廖玉珠、劉明宗、蕭芳蘭、何陽修. 2001. 彩色海芋健康種之微體繁殖與養球技術之建立. 137-149. 健康種苗在植物病害防治上之應用研會專刊. 張雅君、李淑娟、林怡礽. 2005. 番茄叢矮病毒屬快速檢測方法之研發與應用. 植病會刊 14: 41-50. 楊秀珠. 1999. 花卉病害圖鑑. 503. 茂立有限公司出版. 劉明宗、楊佐琦、廖玉珠、蕭芳蘭、何陽修、陳駿季. 2003. 作物簡介及種苗殖與種球生產. 2-7. 植物保護圖鑑系列12 –海芋保護. 行政院農委會動植物防檢疫局出版. 謝廷芳、杜金池. 1995. 百合、彩色海芋之病害與防治。球根花卉產業研討會專輯. 200-212. 農林廳種苗改良繁殖場編印. 台中. Chang, C. H. 2003. Complete genome sequence and genetic organization of Lisianthus necrosis virus, Master thesis, Department of Plant Pathology National Chung Hsing University, Tauchung, Taiwan, R.O.C. 70 pp. Chang, C. H., Yeh, S. D., Chen, C. C., and Jan, F. J. 2007. Complete genome sequence and genetic organization of Lisianthus necrosis virus suggests it should be re-delineated from Necrovirus into Tombusvirus. Arch. Virol. (accepted). Chang, Y. C., Chen, Y. L., and Chung, F. C. 2001. Mosaic disease of calla lily caused by a new potyvirus in Taiwan. Plant Dis. 85: 1289. Chen, C. C., Chang, C. A., Tsai, H. T., Cheng, Y. H., Chao, C. H., and Yeh, S. D. 2000. First detection of turnip mosaic virus on calla lily. Plant Pathol. Bull. 9: 184. Chen, C. C., Chang, C. A., Tsai, H. T., and Hsu, H. T. 2004. Identification of a potyvirus causing latent infection in calla lilies. Plant Dis. 88: 1046. Chen, C. C., Chao, C. H., Chen, C. C., Yeh, S. D., Tsai, H. T., and Chang, C. A. 2003. Identification of Turnip mosaic virus isolates causing yellow stripe and spot on calla lily. Plant Dis. 87: 901-905. Chen, C. C., Chen, T. C., Lin, Y. H., Yeh, S. D., and Hsu, H. T. 2005. A chlorotic spot disease on calla lilies (Zantedeschia spp.) is caused by a tospovirus serologically but distantly related to Watermelon silver mottle virus. Plant Dis.89: 440-445. Chen, C. C, Chen, Y. K., and Hsu, H.T. 2000a. Characterization of a virus infecting lisianthus. Plant Dis. 84: 506-509. Chen, C. C, and Hsu, H.T. 2002. Occurrence of a severe strain of Lisianthus necrosis virus in imported carnation seedlings in Taiwan. Plant Dis. 86: 444. Chen, C. C., Ko, W. F., Jan, F. J., Lin, C. Y., and Hsu, H. T. 2002b. Characterization of Carnation mottle carmovirus isolated from calla lily (Zantedeschia spp.). Plant Pathol. Bull. 11: 242-243. Chen, C. C., Ko, W. F., Jan, F. J., Lin, C. Y., Jan, F. J., and Hsu, H. T. 2003b. First report of Carnation mottle virus in calla lily (Zantedeschia spp.). Plant Dis. 87: 1539. Chen, Y. K., Chen, C. C. and Chen L. C. 1996. Occurrence of powdery mildew on golden calla lily in Taiwan. Ann. Phytopathol. Soc. Jpn. 62: 580-583. Christie, S.R., Purcifull, D.E., Crawford, W.E., and Ahmed, N.A. 1985. Electron microscopy of negatively stained clarified viral concentrates obtained from small tissue samples with appendices on negative staining techniques. Fla. Agr. Sta. Bull.No. 872. 45pp. Chu, M., Desvoyes, B., Turina, M., Noad, R., and Scholthof, K. B. G. 2000. Genetic dissection of tomato bushy stunt virus p19-protein-mediated host-dependent symptom induction and systemic invasion. Virology 266: 79-87. Chu, M., Park, J. W., and Scholthof, H. B. 1999. Separate regions on the tomato bushy stunt virus p22 protein mediate cell-to-cell movement versus elicitation of effective resistance responses. Mol. Plant-Microb. Interact. 12: 285-29. Dijkstra, J., and de Jager, C. P. 1998. Practical Plant Virology - Protocols and Exercises. Springer-Verlag, Berlin. 459pp. Fauquet, C. M., Mayo, M. A., Maniloff, J., Desselberger, U., and Ball, L. A. 2005. Virus Taxonomy. Eighth Report of the International Committee on Taxonomy of Viruses. Academic Press, San Diego. Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783-791. Fuller, M.S. 1978. Lower Fungi in the Laboratory. University of Georgia Press, 190pp. Harlow, E., and Lane, D. 1988. Antibodies – A Laboratory Manual. Cold Spring Harbor Press. 726pp. Hearne, P. Q., Knorr, D. A., Hillman, B., and Morris, T. J. 1990. The complete genome structure and synthesis of infectious RNA frome clones of tomato bushy stunt virus. Virology 177: 141-151. Hsu, S. T., and Tzeng, K. C. 1981. Species of Erwinia associated with soft rot diseases of plants in Taiwan, 9-18 in: Proc. Fifth Int. Conf. Plant Pathol. Bact. Cali., Colombia (J. C. Lozano, ed). Huang, C. H., and Chang, Y. C. 2002. Zantedeschia mild mosaic virus, a new potyvirus infecting callas. Plant Pathol. Bull. 11: 250. Hull, R. 2002. Matthew’ Plant Virology. 4 ed. Academic press. San Diego Iwaki, M., Maria, E.R.A, Hanada, K., and Onogi, S. 1987. Lisianthus necrosis virus, a new necrovirus from Eustoma russellianum. Phytopathology 77: 867-870. Iwaki, M., Maria, E.R.A., Hanada, K., Onogi, S. and Zenbayashi, R. 1985. Three viruses occurred in lisianthus plants. Ann. Phytopathol. Soc. Jpn. 52: 355. Kleinhempel, H., and Kegler, G. 1982. Transmission of Tomato bushy stunt virus without vectors. Acta. Phytopathol. Acad. Sci. Hung. 17: 17-21. Koenig, R. 1981. Indirect ELISA methods for the broad specificity detection of plant viruses. J. Gen. Virol. 55: 53-62. Laemmli , U.K. 1970. Cleavage of structural proteins during the assembly of the ehad of bacteriophage T4. Nature 227: 680-685. Lesemann, D.-E., and Winter, S. 2002. Konjac mosaic virus, Dasheen mosaic virus and unknown potyviruses infecting Zantedeschia spp. and other cultivated Araceae. Acta Hortic. 568: 135-141. Lommel, S. A. , Martelli, G. P., and Russo, M. 2000. Tombusviridae, in: virus Taxonomy: Classification and nomenclature of viruses, (M. H. V. van Regenmortel, C.M. Fauquet, D. H. L. Bishop, E. B. Carsten, M. K. Estes, S. M. Lemon, J. Maniloff, M. A. Mayo, D. j. McGeoch, C. R. Pringle, and R. B. Wickner, Eds.), Academic Press, San Diego. McLean, M. A., Campbell, R. N., Hamilton, R. I., and Rochon, D. M. 1994.Involvement of the Cucumber necrosis virus coat protein in the specificity of fungus transmission by Olpidium bornovanus. Virology 204: 840-842. Morris, T. J. 2001. Tombusviruses, In: Encyclopedia of plant pathology, Inc., New York. Obermeier, C., Sears,J. L., Liu, H. Y., Schlueter, K. O., Ryder, E. J., Duffus, J. E., Koike, S. T., and Wisler, G. C. 2001.Characterization of distinct tombusviruses that cause diseases of lettuce and tomato in the western United States. Phytopathology 91: 797-806. Olson, A. J., Bricogne, G., and Harrison, S. C. 1983. Structure of Tomato bushy stunt virus; IV. The virus particle at 2.9 angstrom resolution. J. Mol. Biol. 171: 61-93. Pham, K., Langeveld, S. A., Lemmers, M. E. C., and Derks, A. F. L. M. 2002. Detection and identification of potyviruses in Zantedeschia. Acta Hortic. 568:143-148. Reynolds, E. S. 1963. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell. Biol. 17: 208-212. Scholthof, H. B., Scholthof, K. B. G., Kikkert, M., and Jackson, A. O. 1995. Tomato bushy stunt virus spread is regulated by two nested genes that function in cell-to-cell movement and host dependent systemic invasion. Virology 213, 425-438. Zettler, F. W., and Hartman, R. D. 1995. Virus diseases of major ornamental crops- Dieffenbachia, Caladium, and Zantedeschia. Pages 464-470 in: Virus-like Diseases of Bulb and Flower Crops. G. Loebenstein, R. H. Lawson, and A. A. Brunt, eds. John Wiley & Sons Inc., Chichester, UK.
摘要: 台灣中部地區田間栽培的彩色海芋 (calla lily, Zantedeschia spp.),發現疑似由病毒感染所引發的系統性壞疽病徵之植株。田間彩色海芋病株呈系統性感染,葉緣捲起,葉面出現黃化斑及壞疽斑,未壞死處略呈嵌紋狀,嚴重者全葉壞疽或整株死亡。罹病葉片粗汁液、純化的懸浮液中及病組織超薄切片,經以電子顯微鏡觀察到一直徑約為 32-34 nm的球形病毒顆粒。間接酵素聯結免疫吸附分析法 (indirect-ELISA)、西方轉漬免疫分析 (western blotting) 試驗均顯示受測的彩色海芋病毒與洋桔梗壞疽病毒 (Lisianthus necrosis virus, LNV) 有血清類緣關係。電泳分析 (SDS-PAGE) 彩色海芋分離株得到一分子量約 39 kDa 的鞘蛋白,此與 LNV 的鞘蛋白分子量相似。進一步針對洋桔梗壞疽病毒彩色海芋分離株 (LNV-Z) 的基因體進行選殖及核酸序列之解析。將LNV-Z 繁殖於其系統性寄主 Nicotiana benthamiana 上,經抽取 LNV-Z 總量 RNA 後,針對其基因體序列,以反轉錄聚合酶鏈鎖反應 (RT-PCR) 進行增幅並選殖所增幅的 cDNA 片段,經解序得知 LNV-Z 基因體全長度為 4785 個核苷酸 (Accession number AM71119)。基因體由 5 個轉譯架構 (ORFs) 所組成,由 5′ 端至 3′ 端分別為 ORF 1 (聚合酶基因,polymerase gene, 2457 nt)、ORF 2 (外鞘蛋白基因,coat protein gene, 1167 nt)、ORF 3 (移動蛋白基因,movement protein gene, 576 nt)、ORF 4 (p19蛋白基因,p19 protein gene, 519 nt)。此基因體結構與 LNV 和其他Tombusvirus 屬病毒相似。序列分析的結果顯示: LNV-Z與 Tombusvirus 屬病毒的鞘蛋白胺基酸相同度 (amino acid identity) 為 36.1~97.7%,其中以與 LNV 洋桔梗分離株 (LNV-L) 的相同度最高(鞘蛋白胺基酸序列相同度 97.7%)。由此分子層次證據進一步顯示引起彩色海芋系統性壞疽病徵的病原確定為洋桔梗壞疽病毒 (LNV)。
Calla lily (Zantedeschia spp.) showing symptoms of systemic necrosis was observed in the fields of central Taiwan in the 2003. Electron microscopic observation revealed the presence of isometric virus particles of 32-34 nm in diameter in the preparations of crude extracts, purified virus suspension and ultrathin sections of diseased tissues. The relative molecular weight of the subunit of viral capsid protein, similar to that of Lisianthus necrosis virus (LNV), was estimated as 39 kDa by SDS-polyacrylamide gel electrophoresis. The isolated virus reacted positively to LNV antiserum in indirect-ELISA and western blotting tests. The cDNA fragments were amplified from the total RNA extracted from LNV-Z infected Nicotiana benthamiana by reverse transcription-polymerase chain reaction (RT-PCR). Full length sequence of the virus genome has been compiled from cDNA fragments amplified by RT-PCR with LNV-specific primer sets. Sequence analysis showed that the complete genome of LNV-Z consists of 4785 nucleotides with five open reading frames (Acc. No. AM71119). The amino acid identity of the coat proteins between tested strain (LNV-Z) and the reported lisianthus strain (LNV-L) was 97.7%. The results shown in this study indicate that LNV is the causal agent of the systemic necrosis of calla lily in the fields.
其他識別: U0005-2208200716274600
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