Abstract
Autophagy is an intracellular process in all eukaryotes which is responsible for the degradation of cytoplasmic constituents, recycling of organelles, and recycling of proteins. It is an important cellular process responsible for the effective virulence of several pathogenic plant fungal strains, having critical impacts on important crop plants including potatoes. However, the detailed physiological mechanisms of autophagy involved in the infection biology of soil-borne pathogens in the potato crop needs to be investigated further. In this study, the autophagy-related gene, FoATG12, in potato dry rot fungus Fusarium oxysporum was investigated by means of target gene replacement and overexpression. The deletion mutant ∆FoATG12 showed reduction in conidial formation and exhibited impaired aerial hyphae. The FoATG12 affected the expression of genes involved in pathogenicity and vegetative growth, as well as on morphology features of the colony under stressors. It was found that the disease symptoms were delayed upon being inoculated by the deletion mutant of FoATG12 compared to the wild-type (WT) and overexpression (OE), while the deletion mutant showed the disease symptoms on tomato plants. The results confirmed the significant role of the autophagy-related ATG12 gene in the production of aerial hyphae and the effective virulence of F. oxysporum in the potato crop. The current findings provid an enhanced gene-level understanding of the autophagy-related virulence of F. oxysporum, which could be helpful in pathogen control research and could have vital impacts on the potato crop.
Highlights
Autophagy is a conserved intracellular degradation and recycling process in eukaryotes which helps to overcome nutrient deficiency and allows cells to survive in starvation conditions
Deletion mutant genes of F. oxysporum strains containing GFP fusion proteins were created by performing replacement strategy in order to reveal the FoATG12 target gene functions (Figure S1)
PCR was used to analyze the hygromycin-resistant (HygR) transformants in which the expected shift in the FoATG12 transformant indicated the successful replacement of the FoATG12 gene (Figure S2)
Summary
Autophagy is a conserved intracellular degradation and recycling process in eukaryotes which helps to overcome nutrient deficiency and allows cells to survive in starvation conditions. The process involves the recycling of degraded products, the synthesis of the macromolecules in vacuole or lysosome, and the formation of autophagosomes an important component of autophagy [1]. About 30 genes have been identified that control autophagy-related cellular mechanism in plants. Among these genes, ATG8 and ATG12 act as the principle components for formation of autophagosomes during nonselective macroautophagy [2]. The deletion mutant of ATG8 and ATG12 showed imperfect autophagy [4]
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