Enterobacter cloacae: a newly identified soft rot pathogen of radish with cross-species pathogenicity

Bacterial soft rot is a major vegetable disease of global significance, predominantly associated with Pectobacterium species; however, new reports indicate that novel, emerging pathogens are contributing to disease incidence. This study identified a novel pathogen, Enterobacter cloacae, as a causal agent of radish soft rot. Two isolates, RDH1 and RDH3, were isolated from 20 decaying radish taproots collected from Kolar, Karnataka, India, where a 12% disease incidence was recorded. Biochemical and physiological characterization, alongside comparison with E. cloacae ATCC 13047, confirmed the genus identity. Molecular analysis of 16S rRNA sequences revealed 99.56 and 99.87% similarity of RDH1 and RDH3, respectively, to known E. cloacae strains. Pathogenicity assay confirmed the pathogenicity of both isolates, and semi-quantitative assessment of plant cell wall degrading enzymes showed RDH1 producing clearance zones of 12.00, 10.33, and 8.00 mm, while RDH3 exhibited zones of 12.00, 10.00, and 7.67 mm, of pectin lyase, polygalacturonase, and cellulase, respectively. Host range assays on 10 vegetable crops revealed RDH3 as more virulent, particularly in radish, carrot, and cabbage, with the hypodermal syringe method showing broader infectivity compared to minimal infection via coir-enrichment seedling inoculation. Further, whole genome sequencing of RDH3 revealed a 4.8 Mb genome, 55% GC content, a single plasmid, and 99% ANI similarity to E. cloacae GGT036, containing T6SS, T4SS, ICEs, prophages, genomic islands, and 12 horizontal gene transfer events. These findings underscore the emerging role of E. cloacae in vegetable soft rot and highlight the need for further research on its pathogenic mechanisms and management strategies.