| 王凯娜, 战斌慧*, 周雪平*.黑龙江地区番茄斑萎病毒的鉴定及其部分生物学特征分析[J].植物保护,2019,45(1):37-43. |
| 黑龙江地区番茄斑萎病毒的鉴定及其部分生物学特征分析 |
| Detection and partial biological characterization of Tomato spotted wilt tospovirus in Heilongjiang Province |
| 投稿时间：2018-03-18 修订日期：2018-05-02 |
| DOI：S 436.412.1 |
| 中文关键词: 番茄斑萎病毒 小RNA深度测序 RT-PCR检测 |
| 英文关键词:Tomato spotted wilt tospovirus small RNA deep sequencing RT-PCR |
| 基金项目:国家自然科学基金青年基金(31701768) |
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| 中文摘要: |
| 近年来, 番茄斑萎病毒Tomato spotted wilt tospovirus (TSWV)在我国多个地区发生严重危害。本研究采用小RNA深度测序和RT-PCR相结合的方法对采自黑龙江的番茄病毒病样品中的病毒进行了鉴定。提取番茄样品的RNA并构建小RNA文库进行测序, 数据分析发现比对至TSWV基因组的reads数占比对到病毒核酸总reads数的92.64%, 表明侵染此番茄样品的病原物可能是TSWV。利用RT-PCR方法进一步确定侵染此番茄样品的病毒为TSWV, 将其命名为TSWV-HLJ1。对TSWV-HLJ1的核苷酸序列进行分析并构建系统进化树, 结果表明TSWV-HLJ1与TSWV云南甜椒分离物(TSWV-YNgp)的亲缘关系最近。介体传毒试验表明, 西花蓟马可将TSWV-HLJ1传播感染健康寄主植物。摩擦接种试验表明, TSWV黑龙江分离物能够侵染本氏烟、辣椒和番茄。这是我国首次报道在黑龙江地区发现TSWV的危害。 |
| 英文摘要: |
| Recently, Tomato spotted wilt tospovirus (TSWV) has been reported in several areas of China causing serious damage. To identify the viruses in tomato samples exhibiting virus-like symptoms from Heilongjiang Province, small RNA deep sequencing and RT-PCR were adopted. Total RNA of tomato samples was extracted and the siRNA library was constructed and sequenced. Data analysis showed that the siRNA reads aligned to TSWV accounted for 92.64 % of the total siRNA reads to the virus sequences deposited in GenBank, indicating that the pathogen infecting the tomato samples might be TSWV. The RT-PCR further confirmed the presence of TSWV, which was named TSWV Heilongjiang isolate (TSWV-HLJ1). The phylogenetic tree based on the whole nucleotide sequence of TSWV isolates indicated that the TSWV-HLJ1 was more closely related to TSWV Yunnan isolate (TSWV-YNgp). Vector transmission experiment showed that Frankliniella occidentalis could acquire TSWV-HLJ1 and transmit it to the healthy plants successfully. Mechanical inoculation experiment showed that TSWV-HLJ1 could infect Nicotiana benthamiana, Capsicum annuum L. and Lycopersicon esculentum Mill. This was the first report of TSWV infecting tomato in Heilongjiang Province. |
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