CRISPR/Cas9-Mediated SlATG5 Mutagenesis Reduces the Resistance of Tomato Fruit to Botrytis cinerea

Abstract

Tomato fruit is highly susceptible to infection by Botrytis cinerea (B. cinerea), a dominant pathogen, during storage. Recent studies have shown that autophagy is essential for plant defense against biotic and abiotic stresses. Autophagy-related gene 5 (ATG5) plays a key role in autophagosome completion and maturation, and is rapidly induced by B. cinerea, but the potential mechanisms of ATG5 in Solanum lycopersicum (SlATG5) in postharvest tomato fruit resistance to B. cinerea remain unclear. To elucidate the role of SlATG5 in tomato fruit resistant to B. cinerea, CRISPR/Cas9-mediated knockout of SlATG5 was used in this study. The results showed that slatg5 mutants were more vulnerable to B. cinerea and exhibited more severe disease symptoms and lower activities of disease-resistant enzymes, such as chitinase (CHI), β-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO), than the wild type (WT). Furthermore, the study observed that after inoculation with B. cinerea, the relative expression levels of genes related to salicylic acid (SA) signaling, such as SlPR1, SlEDS1, SlPAD4, and SlNPR1, were higher in slatg5 mutants than in WT. Conversely, the relative expression levels of jasmonic acid (JA) signaling-related genes SlLoxD and SlMYC2 were lower in slatg5 mutants than in WT. These findings suggested that SlATG5 positively regulated the resistance response of tomato fruit to B. cinerea by inhibiting the SA signaling pathway and activating the JA signaling pathway.