Department of Plant Pathology, Kansas State University, Manhattan, Kansas, USA.
J Virol. 2012 Aug;86(16):8793-809. doi: 10.1128/JVI.00285-12. Epub 2012 Jun 13.
Tomato spotted wilt virus (TSWV) is transmitted by Frankliniella occidentalis in a persistent propagative manner. Despite the extensive replication of TSWV in midgut and salivary glands, there is little to no pathogenic effect on F. occidentalis. We hypothesize that the first-instar larva (L1) of F. occidentalis mounts a response to TSWV that protects it from pathogenic effects caused by virus infection and replication in various insect tissues. A partial thrips transcriptome was generated using 454-Titanium sequencing of cDNA generated from F. occidentalis exposed to TSWV. Using these sequences, the L1 thrips proteome that resolved on a two-dimensional gel was characterized. Forty-seven percent of the resolved protein spots were identified using the thrips transcriptome. Real-time quantitative reverse transcriptase PCR (RT-PCR) analysis of virus titer in L1 thrips revealed a significant increase in the normalized abundance of TSWV nucleocapsid RNA from 2 to 21 h after a 3-h acquisition access period on virus-infected plant tissue, indicative of infection and accumulation of virus. We compared the proteomes of infected and noninfected L1s to identify proteins that display differential abundances in response to virus. Using four biological replicates, 26 spots containing 37 proteins were significantly altered in response to TSWV. Gene ontology assignments for 32 of these proteins revealed biological roles associated with the infection cycle of other plant- and animal-infecting viruses and antiviral defense responses. Our findings support the hypothesis that L1 thrips display a complex reaction to TSWV infection and provide new insights toward unraveling the molecular basis of this interaction.
番茄斑萎病毒（TSWV）通过西方花蓟马以持久增殖的方式传播。尽管 TSWV 在中肠和唾液腺中大量复制，但对西方花蓟马几乎没有致病作用。我们假设西方花蓟马的第一龄幼虫（L1）对 TSWV 产生反应，从而保护其免受病毒感染和复制在各种昆虫组织中引起的致病作用。使用来自暴露于 TSWV 的西方花蓟马的 cDNA 生成的 454-Titanium 测序生成了部分蓟马转录组。使用这些序列，对二维凝胶上解析的 L1 蓟马进行了蛋白质组分析。使用蓟马转录组鉴定了 47%解析出的蛋白斑点。实时定量 RT-PCR（RT-PCR）分析 L1 蓟马中的病毒滴度显示，在感染病毒的植物组织上进行 3 小时获取访问期后 2 至 21 小时，TSWV 核衣壳 RNA 的归一化丰度显着增加，表明感染和病毒积累。我们比较了感染和未感染的 L1 的蛋白质组，以鉴定对病毒显示差异丰度的蛋白质。使用四个生物学重复，有 26 个斑点包含 37 种蛋白质对 TSWV 有明显的变化。这些蛋白质中的 32 个的基因本体分配揭示了与其他植物和动物感染病毒的感染周期以及抗病毒防御反应相关的生物学作用。我们的研究结果支持 L1 蓟马对 TSWV 感染表现出复杂反应的假设，并为揭示这种相互作用的分子基础提供了新的见解。
