洋桔梗上蠶豆萎凋病毒之分離與鑑定

Isolation and Identification of a Fabavirus from Lisianthus

陳慶忠(Ching-Chung Chen) ； 柯文華(Wen-Fwa Ko) ； 曹淑麗(Shu-Li Tsao) ； 趙佳鴻(Chia-Hung Chao)

《臺中區農業改良場研究彙報》 70期 (2001/03) Pp. 51-63

https://doi.org/10.6956/BTDAIS.200103.0051

摘要

自田間洋桔梗（Eustoma russellianum（Don.）Griseb）顯示類似病毒引起之病徵的異常植株分離到一種病毒。罹病洋桔梗於下方葉片出現黃化斑點或組織淡綠至濃綠之多環輪斑，終呈壞疽斑點。除上述病徵外，在其他感受性植物尚可觀察到斑駁、局部壞疽病斑或葉片畸型等徵狀。罹病洋桔梗葉片粗汁液以2%醋酸鈾陰染，於電子顯微鏡可觀察到直徑約28 nm之多角球形病毒。超薄切片洋桔梗、奎蔾、菸草（Nicotiana rustica）及矮牽牛之罹病葉片於葉肉細胞之細胞質內亦可觀察到類似之病毒顆粒，但細胞核則無；罹病奎藜葉片於葉肉細胞之細胞質內可觀察到梯狀構造及不定形含病毒顆粒之內含體。利用高低速交互離心及蔗糖梯度離心純化罹病奎藜或菸草（N. rustica）可得到直徑約28 nm之多角球形病毒顆粒。利用SDS-PAGE電泳分離病毒鞘蛋白，得到兩種蛋白次單位，分子量分別為24及43 kDa。DAS-ELISA及indirect ELISA分析顯示新分離之病毒為Fabavirus成員之蠶豆萎凋病毒－2（Broad bean wilt virus-2）之一系統（strain）。Fabavirus在台灣之發生為首次記錄。

關鍵字

洋桔梗 ； 蠶豆嵌紋病毒 ； 分離 ； 鑑定

並列摘要

A virus was isolated from lisianthus (Eustoma russellianum (Don.) Griseb) showing virus-like disease symptoms. In lisianthus, symptoms initially developed on lower leaves with yellow spots and an irregular concentric ring pattern of dark green and pale green tissues which eventually developed into necrotic concentric rings. In addition to these symptoms, mottling, necrotic local lesions, and sometimes, malformation were observed on leaves of other susceptible plants. Examination of crude sap of infected lisianthus leaves by electron microscopy revealed the presence of spherical particles of 28 nm in diameter when stained with 2% uranyl acetate. Similar particles were present in cytoplasm but not in nuclei in ultrathin sections of leaf tissues from virus-infected plants of Eustoma russellianum, Nicotiana rustica, and petunia hybrida. Cytoplasmic lattice structures and amorphous inclusions containing virions were also observed in ultrathin sections of leaf tissues of infected Chenopodium quinoa. Differential centrifugation followed by sucrose density gradient centrifugation of preparations from tissues of infected C. quinoa or N. rustica yielded virion particles about 28 nm in diameter. Analysis by SDS-polyacrylamide gel electrophoresis of purified virus preparations showed that virions contained two structural polypeptides, with 24 and 43 kDa relative molecular weight, respectively. Results of DAS-ELISA and indirect ELISA indicated that the virus is related to Broad bean wilt virus-2 of the genus Fabavirus.

並列關鍵字

Eustoma russellianum ； Fabavirus ； isolation ； identification