Rhizopus stolonifer Exhibits Necrotrophic Behavior when Causing Soft Rot in Ripe Fruit

Abstract

Rhizopus stolonifer is known for causing soft rot in fruits and vegetables during postharvest. Although it has traditionally been considered a saprophyte, it appears to behave more like a necrotrophic pathogen. In this study, we propose that R. stolonifer invades host tissues by actively killing host cells and overcoming the host defense mechanisms, as opposed to growing saprophytically on decaying plant matter. We tested this hypothesis by characterizing R. stolonifer infection strategies when infecting four fruit hosts (tomato, grape, strawberry, and plum). We started by generating a high-quality genome assembly for R. stolonifer using PacBio sequencing. This led to a genome size of 45.02 Mb, an N50 of 2.87 Mb, and 12,644 predicted loci with protein-coding genes. Next, we performed a transcriptomic analysis to identify genes that R. stolonifer preferentially uses when growing in fruit versus culture media. We categorized these infection-related genes into clusters according to their expression patterns during the interaction with the host. Based on the expression data, we determined that R. stolonifer has a core infection toolbox consisting of strategies typical of necrotrophs, which includes a set of 33 oxidoreductases, 7 proteases, and 4 cell wall-degrading enzymes to facilitate tissue breakdown and maceration across various hosts. This study provides new genomic resources for R. stolonifer and advances the knowledge of Rhizopus–fruit interactions, which can assist in formulating effective and sustainable integrated pest management approaches for soft rot prevention.