甜椒黃化輪紋斑點病之病毒分離與鑑定

Isolation and identification of a virus causing chlorotic ringspot symptom on sweet pepper in Taiwan

姚國山(K. S. Yao) ； 盧耀村(Y. T. Lu)

《植物病理學會刊》 2卷2期 (1993/06) Pp. 57-65

https://doi.org/10.6649/PPB.199306_2(2).0001

摘要

將田間出現黃化輪紋斑點病徵的甜椒葉片接種於紅藜(Chenopodium amaranticolor Coste & Reyn.)，經三次單斑分離後，將病毒回接至藍星甜椒(Capsicum annuum L. cv. Blue-Star).可造成與田間相同之病徵。此病毒經接種於9科35種寄主後，發現能感染藜科 (Chenopodiaceae)、紫花酢醬草科(Oxalidaceae)、茄科(Solanaceae)、豆科(Leguminosae)、莧科(Aizoaceae)和葫蘆科(Cucurbitaceae)等6科26種植物。在Nicotiana glutinosa L.、矮牽牛 (Petunia hybrida Vilm.)及奎藜(C. quinoa willd)上，能引起壞疽病斑和針狀(pin-point)小壞疽病斑，此特性與國外所報告之番茄斑點萎凋病毒辣椒分離株引起之病徵及寄主範圍類似，然而在N. benthamiana Domin.之接種葉所産生的黃化斑點病徵則有所不同。經由Sepharose CL-6B管柱分子篩孔色層分析所得之試液，汁液接種至原寄主測定病原性，以及利用電子顯微鏡觀察，可發現大小在70-120 nm之間，且具有套膜的球形病毒粒子。回接的甜椒病葉經超薄切片後以電子顯微鏡觀察，可在細胞質中發現多數大小與外形相同之球形病毒粒子。此結果顯示本試驗所獲病毒與國外所報告之Bunyaviridae科之Tospovirus屬的病毒特性極爲接近，可能爲相同之病毒。此外，再由免疫分析結果，發現本試驗分離所得病毒與國外報告之番茄斑點萎凋病毒的萵苣系統(tomato spotted wilt virus-lettuce, TSWV-L)血清具有陽性反應，而與鳳仙花系統(TSWV-I)無反應，因此推測本試驗分離所得之病毒，應屬於番茄斑點萎凋病毒萵苣系統之台灣分離株。

關鍵字

甜椒 ； 番茄斑點萎凋病毒

並列摘要

Sweet pepper plants showing chlorotic ringspot symptom were collected from on field in Taichung area in 1991. A virus isolate was obtained by three successive single lesion Chenopodium amaranticolor. To determine the host range, back-inoculated Capsicum annuum L cv. Blue-Star was used as virus inoculum and inoculated to 35 plant species of 9 families. The results showed that the virus infected 26 plant species in Chenopodiaceae, Oxalidaceae Solanaceae, Leguminosae, Aizoaceae, and Cucurbitaceae Chlorotic or pin-point local lesions were observed on Nicotiana glutinosa, Petunia hybrida, and C. quinoa. The host range and reactions were similar to those of the reported pepper isolate of tomato spotted wilt virus (TSWV-P). The virus however caused chlorotic spots on inoculated leaves of N. benthamiana, distinct from TSWV-P isolate. The virus was partially purified by Sepharose CL-6B column chromatography, and then eluted with 0.02 M potassium phosphate buffer containing 0.01 M EDTA, and 0.1% (W/V) L-cysteine, pH 7.0. Virus infectivity was confirmed by back inoculation to pepper plants. Electron microscopy of negatively stained purified virus or virus particles present in ultrathin sections from leaves of diseased pepper revealed that the morphology of virus was roughly enveloped spherical with diameter ranging from 70-120 nm, indicating that it belongs to Tospovirus genus of arthropod-borne Bunyaviridae. In addition, serological tests by enzyme linked immunosorbent assay with polyclonal antibodies against different serotypes of tospovirus demonstrated that the virus was serologically related to the L-serotype of Tospovirus. Evidences from this study suggest that the virus inducing chlorotic ringspot on sweet pepper is a new isolate of TSWV previously not found in Taiwan.

並列關鍵字

Sweet pepper ； Tomato spotted wilt virus ； Tospovirus