Table_3_A Novel Tomato Fusarium Wilt Tolerance Gene.XLSX

The reduced mycorrhizal colonization (rmc) tomato mutant is unable to form mycorrhiza and is more susceptible to Fusarium wilt compared with its wild-type isogenic line 76R. The rmc mutant has a chromosomal deletion affecting five genes, one of which is similar to CYCLOPS. Loss of this gene is responsible for non-mycorrhizality in rmc but not enhanced Fusarium wilt susceptibility. Here, we describe assessment of a second gene in the rmc deletion, designated Solyc08g075770 that is expressed in roots. Sequence analyses show that Solyc08g075770 encodes a small transmembrane protein with putative phosphorylation and glycosylation sites. It is predicted to be localized in the plasma membrane and may function in transmembrane ion transport and/or as a cell surface receptor. Complementation and knock-out strategies were used to test its function. Some putative CRISPR/Cas-9 knock-out transgenic events exhibited Fusarium wilt susceptibility like rmc and some putative complementation lines were 76R-like, suggesting that the tomato Solyc08g075770 functions in Fusarium wilt tolerance. This is the first study to demonstrate that Solyc08g075770 is the contributor to the Tfw locus, conferring tolerance to Fusarium wilt in 76R which was lost in rmc.菌根定殖减弱（reduced mycorrhizal colonization, rmc）番茄突变体无法形成菌根，且相较于其野生型同基因系76R，更易感染镰刀菌枯萎病（Fusarium wilt）。该rmc突变体存在一处染色体缺失，涉及5个基因，其中一个基因与CYCLOPS同源。该基因的缺失是rmc突变体无法形成菌根的原因，但并非其镰刀菌枯萎病易感性升高的诱因。本研究对rmc缺失区域内的第二个基因——命名为Solyc08g075770，该基因在根系中表达——进行了功能评估。序列分析表明，Solyc08g075770编码一种小型跨膜蛋白，带有预测的磷酸化和糖基化位点。该蛋白被预测定位于质膜，可能参与跨膜离子转运，或作为细胞表面受体发挥功能。研究通过互补实验与基因敲除策略对其功能进行了验证。部分潜在的CRISPR/Cas-9基因敲除转基因株系表现出与rmc相似的镰刀菌枯萎病易感性，而部分潜在的互补转化株系则与76R表型一致，这表明番茄Solyc08g075770参与调控镰刀菌枯萎病的耐受性。本研究首次证实，Solyc08g075770是Tfw位点的功能基因，该基因赋予76R镰刀菌枯萎病耐受性，而该耐受性在rmc突变体中已丧失。