Purification of Tomato Spotted Wilt Virus Isolated from Tobacco Plants in Morioka, Japan
Purification of Tomato Spotted Wilt Virus Isolated from Tobacco Plants in Morioka, Japan
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从日本盛冈烟草植物中分离的番茄斑萎病毒的纯化
DOI：
10.3186/jjphytopath.57.239
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
K. Tomaru
中科院分区：
文献类型：
--
作者：
S. Tsuda;K. T. Natsuaki;K. Tomaru
从盛冈市的烟草植物中分离出的病毒被确认为番茄斑萎病毒（TSWV），并命名为 TSWV-M 分离株。该病毒在烟草、番茄和本塞姆氏烟草等多种植物上表现出严重症状。 TSWV-M 用于纯化研究。在用于提取 TSWV 的七种缓冲液中，含有 0.01M Na2SO3 和 0.01M EDTA·2Na 的 0.1M 磷酸盐缓冲液是合适的，通过使用 N. tabacum cv. 的半叶法检查粗汁液的感染性较高。克桑西公司在浓度为 15.0% 至 60.0% (w/w) 的等密度线性蔗糖密度梯度的纯化方法中，TSWV 感染性的峰值分数显示为 42.0% (w/w) 蔗糖分数。然而，通过这种方法，主体成分不能从峰分数中消除。在等密度逐步蔗糖密度梯度法中，陷阱为40.0%（w/w），宿主成分可以从具有高感染性的42.0%（w/w）蔗糖组分中排除。通过电子显微镜在级分中观察到完整的TSWV颗粒。随着TSWV纯化过程的通过，最后一步的比感染力计算为OD260的44倍和OD280的46倍，与第一步制备的比率高46倍。
A virus isolated from tobacco plants in Morioka was confirmed as tomato spotted wilt virus (TSWV) and designated as TSWV-M isolate. The virus shows severe symptoms on several plants including tobacco, tomato and Nicotiana benthamiana. TSWV-M was used for the purification study. Out of seven kinds of buffers for extraction of TSWV, 0.1M phosphate buffer containing 0.01M Na2SO3 and 0.01M EDTA⋅2Na was suitable with high infectivity in crude sap checked by half-leaf method using N. tabacum cv. Xanthi nc. In a purification method with isopycnic linear sucrose density gradient with concentration from 15.0% to 60.0% (w/w), the peak fraction of TSWV infectivity was indicated at 42.0% (w/w) sucrose fraction. By this method, however, the host components could not be eliminated from the peak fraction. In isopycnic step-wise sucrose density gradient method with trap of 40.0% (w/w), host components could be excluded from 42.0% (w/w) sucrose fraction with high infectivity. Intact TSWV particles were observed in the fraction by the electron microscopy. With the passage of the TSWV purification process, specific infectivities in the final step were calculated as 44 times at OD260 and 46 times higher at OD280 in the ratio to the first step preparation.