Phytopathology. 2001 Sep;91(9):856-63. doi: 10.1094/PHYTO.2001.91.9.856.
ABSTRACT To clarify the serological relationship of Peanut chlorotic fan-spot virus (PCFV) with other tospoviruses, antisera were produced against the nucleocapsid (N) proteins of this virus and tospoviruses from four serogroups including Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV), Groundnut ringspot virus (GRSV), and Watermelon silver mottle virus (WSMoV). In immunodiffusion tests, the antisera only reacted with their homologous antigens. Similar results were noticed in indirect enzyme-linked immunosorbent assay and immunoblot tests, with the exception that strong cross-reactions were observed in heterologous combinations between TSWV and GRSV. The results indicated that the N protein of PCFV is not serologically related to those of the tospoviruses from the four serogroups. To further characterize the virus, viral S double-stranded RNA was extracted from PCFV-infected Chenopodium quinoa and used for cDNA cloning and sequencing. The full-length viral strand of the S RNA was determined to be 2,833 nucleotides, with an inverted repeat at the 5' and 3' ends and two open reading frames in an ambisense arrangement. The 3'-terminal sequence (5'-AUUGCUCU-3') of the viral S RNA is identical to those of other tospoviruses, indicating that PCFV belongs to the genus Tospovirus. The N and the NSs proteins of PCFV share low amino acid identities (22.3 to 67.5% and 19.3 to 54.2%) with those of reported tospoviruses, respectively. The phylogenetic dendrogram of the N gene of PCFV compared with those of other tospoviruses indicates that PCFV is distinct from other tospoviruses. In hybridization analyses, an N gene cDNA probe of PCFV did not react with viral RNAs of TSWV, GRSV, INSV, and WSMoV, and vice versa. Thus, based on these results, we conclude that PCFV is a new tospovirus species.
摘要 为了阐明花生褪绿斑驳病毒（PCFV）与其他弹状病毒的血清学关系，我们针对该病毒和来自四个血清群的弹状病毒（包括番茄斑萎病毒（TSWV）、矮牵牛坏死斑点病毒（INSV）、花生环斑病毒（GRSV）和西瓜银斑驳病毒（WSMoV））的核衣壳（N）蛋白产生了抗血清。在免疫扩散试验中，抗血清仅与同源抗原发生反应。间接酶联免疫吸附试验和免疫印迹试验也得到了类似的结果，除了 TSWV 和 GRSV 之间的异源组合中观察到强烈的交叉反应。结果表明，PCFV 的 N 蛋白与来自四个血清群的弹状病毒在血清学上没有关系。为了进一步表征该病毒，我们从 PCFV 感染的藜麦中提取了病毒 S 双链 RNA，用于 cDNA 克隆和测序。S RNA 的全长病毒链被确定为 2833 个核苷酸，在 5'和 3'末端有一个反向重复序列，并且以反义排列有两个开放阅读框。病毒 S RNA 的 3'末端序列（5'-AUUGCUCU-3'）与其他弹状病毒相同，表明 PCFV 属于 Tospovirus 属。PCFV 的 N 和 NSs 蛋白与报道的弹状病毒的氨基酸同一性分别为 22.3%至 67.5%和 19.3%至 54.2%。PCFV 的 N 基因与其他弹状病毒的系统发育树表明，PCFV 与其他弹状病毒不同。在杂交分析中，PCFV 的 N 基因 cDNA 探针与 TSWV、GRSV、INSV 和 WSMoV 的病毒 RNA 均不发生反应，反之亦然。因此，基于这些结果，我们得出结论，PCFV 是一种新的弹状病毒种。
