Functional identification of two minor capsid proteins from Chinese wheat mosaic virus using its infectious full-length cDNA clones
Functional identification of two minor capsid proteins from Chinese wheat mosaic virus using its infectious full-length cDNA clones
复制标题
利用其感染性全长 cDNA 克隆对中国小麦花叶病毒的两种次要衣壳蛋白进行功能鉴定
DOI：
10.1099/jgv.0.000532
复制
发表时间：
2016-09-01
影响因子：
3.8
通讯作者：
Zhang, Heng-Mu
中科院分区：
文献类型：
--
作者：
Yang, Jian;Zhang, Fen;Zhang, Heng-Mu
通过单次反转录PCR反应获得了中国小麦花叶病毒（CWMV）RNA 1和RNA 2的全长cDNA克隆，其转录本在小麦和本氏烟草（Nicotianabenthamiana）中均具有侵染性。建立了一种高效、可靠的土壤渗透法用于烟草的反向遗传分析。本萨米亚纳感染的cDNA克隆接种导致明显的褪绿症状，CWMV病毒基因组RNA，衣壳蛋白（CP）相关蛋白，和典型的杆状颗粒检测接种和上部叶片，类似于WT病毒。病毒增殖的最适温度为12 ℃，但在植物中系统感染的最适温度为17 ℃。突变体克隆，废除的N-或C-末端延伸的主要CP不抑制系统性感染或形成杆状颗粒，但有时修改接种植物的症状。这些结果表明，CWMV的两个次要CP相关蛋白与病毒在植物中的感染、复制、系统运动和病毒体组装有关。
Full-length cDNA clones of Chinese wheat mosaic virus (CWMV) RNA1 and RNA2 were produced from single reverse transcription PCR reactions and transcripts were shown to be infectious in both wheat and Nicotiana benthamiana. An efficient and reliable agro-infiltration method was then developed for reverse genetic assays in N. benthamiana. Inoculation of infectious cDNA clones resulted in obvious chlorotic symptoms, and CWMV viral genomic RNAs, capsid protein (CP)-related proteins, and typical rod-shaped particles were detectable on the inoculated and upper leaves, similar to those of WT virus. The optimal temperature for virus multiplication was 12 degrees C, but the optimum for systematic infection in plants was 17 degrees C. Mutant clones that abolished the N- or C-terminal extensions of the major CP did not inhibit systemic infection or the formation of rod-shaped particles but sometimes modified the symptoms in inoculated plants. These results suggest that the two minor CP-related proteins of CWMV are dispensable for viral infection, replication, systemic movement and virion assembly in plants.