感染地黃(Rehmannia glutinosa)菸草嵌紋病毒(Tobacco mosaic virus)之鑑定

廖吉彥(J. Y. Liao) ； 胡仲祺(C. C. Hu) ； 高瑞隆(J. L. Kao) ； 鄧汀欽(T. C. Deng)

《植物病理學會刊》 16卷2期 (2007/06) Pp. 61-69

https://doi.org/10.6649/PPB.200706_16(2).0002

摘要

從埔里地黃(Rehmannia glutinosa Libosch.)栽種區及本所中藥材栽培區採集各種疑似病毒感染的植株，汁液接種經單斑分離得到一純系病毒株(RG-1)。從寄主範圍試驗、電子顯微鏡觀察、凝膠雙擴散反應、電泳分析、西方轉漬反應及核酸與胺基酸序列比較，證實所分離的病毒株RG-1係屬於菸草嵌紋病毒(Tobacco mosaic virus, TMV)的一個獨立系統(strain)。本病毒接種奎藝(Chenopodium quinoa)後會產生局部壞疽病斑。經陰染法電子顯微鏡觀察病毒顆粒為長桿狀，大小約300-310×16-18 nm。病毒經接種至奎蔡繁殖，再利用硫酸銫等密度平衡離心法純化，純化所得試料為抗原用以注射白兔製備抗血清。電泳分析純化的病毒鞘蛋白，其主要蛋白分子量約為19 kDa。經西方轉漬反應、SDS-瓊脂免疫擴散反應(immunodiffusion)，自製的抗體與RG-1有同源(homologous)血清反應。從本病毒RNA 3'端解序480個核苷酸，與GenBank中的TMV(Access No．AY555269)比較鞘蛋白基因之核苷酸，其相同度達97.1%，而其肢基酸相似度則為99.4%。本報告為台灣地區TMV感染地黃植株的首篇紀錄。

關鍵字

地黃 ； 菸草嵌紋病毒 ； 病徵 ； 抗血清 ； 比對

並列摘要

Rehmannia glutinosa being a kind of medicinal plant cultivated in Taiwan was found to be infected by viruses. A plant with leaf mosaic was collected from Puli, and from which a pure line of virus isolate (RG1) was obtained through series of single lesion isolation. By host range test, electron microscope observation, immunosorbent electron microscopy, Ouchterlony immunodiffusion, coat protein electrophoresis, western blot analysis, and nucleotide sequencing, and analyses, the virus was identified as a isolate of Tobacco mosaic virus (TMV). The virus systemically infects Rehmannia glutinosa and causes mosaic symptom, but locally infects Chenopodium amaranticolor, C. quinoa and Nicotiana tabacum cv. Vam-Hicks and caused necrotic lesions. Direct negative staining by electron microscopy observation of diseased tissue revealed the presence of rod-shaped virus particles, around 300-310×16-18 nm. The virus was purified from inoculated leaves of C. quinoa by isopycnic centrifugation in cesium sulfate and the purified preparation was used to inject rabbit for antiserum production. The virus was found consisting of one species of coat protein subunit with an estimated molecular weight of 19 kDa. In Ouchterlony immunodiffusion and western blot analyses, the prepared antiserum reacted with homologous TMV antigens. The 3'-terminal 1044 nucleotides of this virus were cloned, sequenced and compared with corresponding sequences of representative Tobacco mosaic virus strains listed in public databases. It was found that this virus is most closely related to the Tobacco mosaic virus reported in China (Accession No. AY555269), with nucleotide and amino acid sequence identities of 97.1% and 99.4%, respectively. This is the first report of Tobacco mosaic virus infecting Rehmannia glutinosa in Taiwan.

並列關鍵字

Rehmannia glutinosa ； Tobacco mosaic virus ； symptom ； antiserum ； sequence comparison