Tomato chlorosis virus (ToCV) causes mild to severe chlorosis on tomato leaves. Although it has been postulated that chlorosis is caused by changes in biosynthesis, degradation, function and/or structure of chlorophyll, mechanisms of virus-induced chlorosis has not been explored to date. To understand the mechanisms of virus-induced chlorosis, ToCV-tomato interaction was studied at transcriptome level using suppression subtractive hybridization (SSH) followed by macroarray hybridization and RT-qPCR. First, a susceptible tomato cultivar, MoneyMaker was graft-inoculated with ToCV. Next, cDNAs were synthesized from mRNAs of leaf samples from ToCV-inoculated and mock-inoculated control plants at 30-day of post inoculation (dpi). Then, a cDNA library enriched with ToCV-repressed genes was constructed using SSH. 250 randomly selected clones from library were sequenced and the expressions of 140 genes were determined by nylon macroarray hybridizations. The expression of 119 genes were significantly decreased by 1.5 or greater fold in response to ToCV infection. The expressions of many genes involved in transcription, defense response, metabolism, and photosynthesis, signaling and protein fate were repressed upon infection with ToCV. The genes repressed by ToCV in tomato were mostly associated with organelles, especially with the chloroplast. After expression analysis of 40 selected ToCV-repressed genes by RT-qPCR assay, the repression of 37 genes in response to ToCV infection was confirmed. Furthermore, a detail expression analysis of ferredoxin 2 (Fd2) gene between 0 dpi and 45 dpi in ToCV-inoculated and mock-inoculated control plants showed that the expression of Fd-2 gene was significantly repressed at 8 to 30 dpi in ToCV-infected plants.

番茄萎黄病毒(ToCV) 导致番茄叶片轻度至重度萎黄。尽管已经假设黄化是由叶绿素的生物合成、降解、功能和/或结构的变化引起的，但迄今为止尚未探索病毒诱导的黄化机制。为了了解病毒引起的萎黄病的机制，在转录组水平上使用抑制消减杂交 (SSH) 和宏阵列杂交和 RT-qPCR 研究了 ToCV 与番茄的相互作用。首先，对易感番茄品种 MoneyMaker 进行了嫁接接种 ToCV。接下来，从接种后 30 天 (dpi) 的 ToCV 接种和模拟接种对照植物的叶样品的 mRNA 合成 cDNA。然后，使用 SSH 构建了富含 ToCV 抑制基因的 cDNA 文库。从文库中随机选择的250个克隆被测序，并通过尼龙宏阵列杂交确定140个基因的表达。响应 ToCV 感染，119 个基因的表达显着降低了 1.5 倍或更多。感染 ToCV 后，许多参与转录、防御反应、代谢和光合作用、信号传导和蛋白质命运的基因的表达受到抑制. 番茄中被 ToCV 抑制的基因主要与细胞器有关，尤其是与叶绿体有关。通过 RT-qPCR 测定对 40 个选定的 ToCV 抑制基因的表达分析后，证实了响应 ToCV 感染的 37 个基因的抑制。此外，在接种 ToCV 和模拟接种的对照植物中，铁氧还蛋白 2 ( Fd2 ) 基因在 0 dpi 和 45 dpi 之间的详细表达分析表明，在 ToCV 感染的对照植物中，Fd-2基因的表达在 8 到 30 dpi 受到显着抑制。植物。