Less is more: CRISPR/Cas9-based mutations in DND1 gene enhance tomato resistance to powdery mildew with low fitness costs

作者：Ruiling Li, Lei Cui, Matteo Martina, Valentina Bracuto, Fien Meijer‐Dekens, Anne‐Marie A. Wolters, Andrea Moglia, Yuling Bai, Alberto Acquadro · 发表于：BMC Plant Biology · 年份：2024 · DOI：10.1186/s12870-024-05428-3 · 被引用次数：17 · 研究领域：Plant Virus Research Studies、Plant-Microbe Interactions and Immunity、Powdery Mildew Fungal Diseases

Abstract Powdery mildew (PM), triggered by Oidium neolycopersici , represents a significant threat and a major concern for the productivity of tomato plants ( Solanum lycopersicum L.). The presence of susceptibility (S) genes in plants facilitates pathogen proliferation and their dysfunction can lead to a recessively inherited broad-spectrum and durable type of resistance. Past studies have demonstrated that disrupting the function of DND1 (Defense No Death 1) increases plant resilience against various pathogens, such as powdery mildew (PM), but this comes at the cost of negatively affecting the overall health and vigor of the plant. To investigate the possibility of minimizing the adverse effects of the dnd 1 mutation while boosting disease resistance, a CRISPR-Cas9 construct with four single guide RNAs targeting three exons of SlDND1 (Solyc02g088560.4.1) was designed and introduced into the tomato variety Moneymaker (MM) through Agrobacterium tumefaciens -mediated transformation. Three T 1 lines (named E1, E3 and E4) were crossed with MM and then selfed to produce T F2 families. All the T F2 plants in homozygous state dnd1/dnd1 , showed reduced PM symptoms compared to the heterozygous ( DND1/dnd1 ) and wild type ( DND1/DND1) ones. Two full knock-out (KO) mutant events (E1 and E4) encoding truncated DND1 proteins, exhibited clear dwarfness and auto-necrosis phenotypes, while mutant event E3 harbouring deletions of 3 amino acids, showed normal growth in height with less auto-...