Key Lab of Integrated Crop Pest Management of Shandong Province, College of Plant Health and Medicine, Qingdao Agricultural University, Qingdao 266109, China.
Int J Mol Sci. 2022 Jun 11;23(12):6541. doi: 10.3390/ijms23126541.
Tomato chlorosis virus (ToCV) has seriously impacted tomato production around the world. ToCV is semi-persistently transmitted by the whitefly, , which is a serious agricultural pest in the world. However, the interaction mechanism between ToCV and its whitefly vector is still poorly understood. Our previous transcriptome analysis demonstrated that the expression level of an immune-related gene, prophenoloxidase (PPO), in increased after ToCV acquisition, which indicates that the PPO may be involved in the interaction mechanism between the ToCV and its vector. To determine the role of the PPO in the acquisition and retention of ToCV by , we cloned the complete Open Reading Frames (ORF) of the s ( and ), and then structure and phylogenetic analyses were performed. s were closely related to the PPO genes of Hemiptera insects. Spatial-temporal expression detection was qualified by using reverse transcription quantitative PCR (RT-qPCR), and this revealed that s were expressed in all tissues and developmental stages. We found that only was significantly upregulated after acquired ToCV for 12 and 24 h. According to the paraffin-fluorescence probe-fluorescence in situ hybridization (FISH) experiment, we verified that ToCV and were co-located in the thorax of , which further revealed the location of their interaction. Finally, the effects of the s on ToCV acquisition and retention by were determined using RNA interference (RNAi). The results showed that the RNAi of the responsive gene () significantly increased the titer of ToCV in . These results demonstrate that participates in ToCV acquisition and retention by .
番茄褪绿病毒（ToCV）已严重影响全球番茄生产。ToCV 由烟粉虱半持久传播，烟粉虱是世界范围内严重的农业害虫。然而，ToCV 与其粉虱介体之间的互作机制仍知之甚少。我们之前的转录组分析表明，感染 ToCV 后 中与免疫相关的基因原酚氧化酶（PPO）的表达水平增加，这表明 PPO 可能参与了 ToCV 与其介体的互作机制。为了确定 PPO 在 获得和保留 ToCV 中的作用，我们克隆了 s（和 ）的完整开放阅读框（ORF），然后进行了结构和系统发育分析。s 与半翅目昆虫的 PPO 基因密切相关。通过反转录定量 PCR（RT-qPCR）进行时空表达检测，结果表明 s 在所有组织和发育阶段均有表达。我们发现，只有 在感染 ToCV 12 和 24 小时后才显著上调。根据石蜡荧光探针-荧光原位杂交（FISH）实验，我们验证了 ToCV 和 均位于 的胸部，这进一步揭示了它们相互作用的位置。最后，通过 RNA 干扰（RNAi）测定了 s 对 的 ToCV 获得和保留的影响。结果表明，响应基因（）的 RNAi 显著增加了 中 ToCV 的滴度。这些结果表明 参与了 ToCV 的获得和保留。
