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通过对其双链 RNA 的部分特征分析鉴定甘蔗条纹花叶病毒。

Identification of sugarcane striate mosaic-associated virus by partial characterization of its double-stranded RNA.

出版信息

Phytopathology. 1999 Oct;89(10):877-83. doi: 10.1094/PHYTO.1999.89.10.877.

Abstract

ABSTRACT Sugarcane striate mosaic (ScSM)-affected sugarcane leaves contain a disease-associated 9-kilobase (kb) double-stranded RNA (dsRNA), usually together with 6- and 2.6-kb dsRNAs. The purified 9-kb dsRNA was amplified by the randomly primed polymerase chain reaction (PCR) and cloned. The nucleotide sequences of three separate regions, representing about 2.55 kb (28%) of the dsRNA sequence, were found to have significant similarities to viruses in the genera Capillo-, Carla-, Fovea-, Potex-, Poty-, Tricho-, and Tymovirus. Greatest overall similarity was found to apple stem pitting virus, with less similarity to blueberry scorch virus and potato virus M. A standard virus purification procedure was used to identify slightly flexuous filamentous particles that copurified with the disease-associated RNA. Particle modal lengths were approximately 950 and 1,900 nm with a diameter of 15 nm. Preparations contained a 51-kDa putative capsid protein and a 9-kb single-stranded RNA with a probable 3' polyadenylate tract. These ScSM-associated virus particles differ physically from viruses in existing genera because of their relative rigidity, length, and putative coat protein size. Reverse-transcription PCR with a primer pair designed from the sequenced segments amplified a 820-base pair fragment from ScSM-affected but not healthy sugarcane plants.

摘要

摘要 受甘蔗条纹花叶病(ScSM)影响的甘蔗叶片含有一种与疾病相关的 9 千碱基(kb)双链 RNA(dsRNA),通常还含有 6-kb 和 2.6-kb dsRNA。通过随机引物聚合酶链反应(PCR)扩增并克隆纯化的 9-kb dsRNA。三个独立区域的核苷酸序列,代表约 2.55 kb(28%)的 dsRNA 序列,与 Capillo-、Carla-、Fovea-、Potex-、Poty-、Tricho-和 Tymovirus 属的病毒具有显著相似性。与苹果茎陷点病毒的总体相似性最大,与蓝莓焦枯病毒和马铃薯病毒 M 的相似性较小。采用标准病毒纯化程序鉴定与疾病相关 RNA 共纯化的略微弯曲丝状颗粒。颗粒的模态长度约为 950nm 和 1900nm,直径为 15nm。制剂中含有 51kDa 的假定衣壳蛋白和 9kb 的单链 RNA,可能具有 3' 多聚腺苷酸尾。由于其相对刚性、长度和假定的外壳蛋白大小,这些与 ScSM 相关的病毒颗粒在物理上与现有属中的病毒不同。用从测序片段设计的引物对进行逆转录 PCR 从受 ScSM 影响但不受健康甘蔗植物扩增出 820 碱基对片段。

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