Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València), 46022 Valencia, Spain.
Centre for Research in Agricultural Genomics (CSIC-IRTA-IAB-UB), Bellaterra, Barcelona, Spain.
J Exp Bot. 2023 Mar 13;74(5):1564-1578. doi: 10.1093/jxb/erac361.
Potato spindle tuber viroid (PSTVd) is a plant pathogen naturally infecting economically important crops such as tomato (Solanum lycopersicum). Here, we aimed to engineer tomato plants highly resistant to PSTVd and developed several S. lycopersicum lines expressing an artificial microRNA (amiRNA) against PSTVd (amiR-PSTVd). Infectivity assays revealed that amiR-PSTVd-expressing lines were not resistant but instead hypersusceptible to the viroid. A combination of phenotypic, molecular, and metabolic analyses of amiRNA-expressing lines non-inoculated with the viroid revealed that amiR-PSTVd was accidentally silencing the tomato STEROL GLYCOSYLTRANSFERASE 1 (SlSGT1) gene, which caused late developmental and reproductive defects such as leaf epinasty, dwarfism, or reduced fruit size. Importantly, two independent transgenic tomato lines each expressing a different amiRNA specifically designed to target SlSGT1 were also hypersusceptible to PSTVd, thus demonstrating that down-regulation of SlSGT1 was responsible for the viroid-hypersusceptibility phenotype. Our results highlight the role of sterol glycosyltransferases in proper plant development and indicate that the imbalance of sterol glycosylation levels favors viroid infection, most likely by facilitating viroid movement.
马铃薯纺锤块茎类病毒(PSTVd)是一种天然感染重要经济作物(如番茄)的植物病原体。在这里,我们旨在培育对 PSTVd 高度抗性的番茄植株,并开发了几种表达针对 PSTVd 的人工 microRNA(amiRNA)的番茄品系(amiR-PSTVd)。感染性测定表明,表达 amiR-PSTVd 的品系对病毒没有抗性,而是超敏。对未接种病毒的 amiRNA 表达系进行表型、分子和代谢分析表明,amiR-PSTVd 意外地沉默了番茄甾醇糖基转移酶 1(SlSGT1)基因,导致叶片内卷、矮化或果实变小等发育和生殖后期缺陷。重要的是,两个独立的转基因番茄品系,每个品系都表达了一种不同的针对 SlSGT1 的特异性 amiRNA,它们也对 PSTVd 表现出超敏性,因此表明 SlSGT1 的下调是导致病毒超敏性表型的原因。我们的结果强调了甾醇糖基转移酶在植物正常发育中的作用,并表明甾醇糖基化水平的失衡有利于病毒的感染,这很可能是通过促进病毒的运动。