Department of Stress Biology and Plant Pathology, CEBAS-CSIC, Campus Universitario de Espinardo, 30100, Murcia, Spain.
Department of Plant Breeding, CEBAS-CSIC, Campus Universitario de Espinardo, 30100, Murcia, Spain.
New Phytol. 2023 Apr;238(1):332-348. doi: 10.1111/nph.18728. Epub 2023 Feb 4.
Pepino mosaic virus (PepMV) is pandemic in tomato crops, causing important economic losses world-wide. No PepMV-resistant varieties have been developed yet. Identification of host factors interacting with PepMV proteins is a promising source of genetic targets to develop PepMV-resistant varieties. The interaction between the PepMV coat protein (CP) and the tomato glutathione S-transferase (GST) SlGSTU38 was identified in a yeast two-hybrid (Y2H) screening and validated by directed Y2H and co-immunoprecipitation assays. SlGSTU38-knocked-out Micro-Tom plants (gstu38) generated by the CRISPR/Cas9 technology together with live-cell imaging were used to understand the role of SlGSTU38 during infection. The transcriptomes of healthy and PepMV-infected wild-type (WT) and gstu38 plants were profiled by RNA-seq analysis. SlGSTU38 functions as a PepMV-specific susceptibility factor in a cell-autonomous manner and relocalizes to the virus replication complexes during infection. Besides, knocking out SlGSTU38 triggers reactive oxygen species accumulation in leaves and the deregulation of stress-responsive genes. SlGSTU38 may play a dual role: On the one hand, SlGSTU38 may exert a proviral function depending on its specific interaction with the PepMV CP; and on the other hand, SlGSTU38 may delay PepMV-infection sensing by participating in the redox intracellular homeostasis in a nonspecific manner.
Pepino mosaic virus (PepMV) 在番茄作物中流行,在全球范围内造成了重大的经济损失。目前还没有培育出抗 PepMV 的品种。鉴定与 PepMV 蛋白相互作用的宿主因子是开发抗 PepMV 品种的遗传靶标的有前途的来源。在酵母双杂交 (Y2H) 筛选中鉴定了 PepMV 外壳蛋白 (CP) 与番茄谷胱甘肽 S-转移酶 (GST) SlGSTU38 之间的相互作用,并通过定向 Y2H 和共免疫沉淀实验进行了验证。SlGSTU38 被 CRISPR/Cas9 技术敲除的 Micro-Tom 植物 (gstu38) 与活细胞成像一起用于了解 SlGSTU38 在感染过程中的作用。通过 RNA-seq 分析对健康和 PepMV 感染的野生型 (WT) 和 gstu38 植物的转录组进行了分析。SlGSTU38 作为一种 PepMV 特异性易感性因子,以细胞自主的方式发挥作用,并在感染过程中重新定位到病毒复制复合物。此外,敲除 SlGSTU38 会在叶片中引发活性氧物质的积累,并导致应激响应基因的失调。SlGSTU38 可能发挥双重作用:一方面,SlGSTU38 可能通过与 PepMV CP 的特异性相互作用发挥助病毒功能;另一方面,SlGSTU38 可能通过参与非特异性的细胞内氧化还原内稳态来延迟 PepMV 感染的感应。