Engineering a novel entomopathogenic strain Pseudomonas chlororaphis for efficient production of double‐stranded RNAs and pest control

RNA interference (RNAi)-based pesticides are emerging as the next generation of pest control solutions. We have previously identified Pseudomonas chlororaphis strain R3-3, which exhibits toxicity towards Plagiodera versicolora. We engineered a mutant strain derived from R3-3, named P. chlororaphis BM, through knocking out the RNase III gene and incorporating a T7 RNA polymerase expression system to boost dsRNA production. We revealed that P. chlororaphis BM produced comparable amounts of dsRNA to the strain E. coli HT115 (DE3) while maintaining its insecticidal activity. Importantly, insect feeding bioassays demonstrated that P. chlororaphis BM expressing dsRNA targeted β-Actin (encoding β-actin protein) of P. versicolora and Srp54K (encoding signal recognition particle protein 54 k) of Henosepilachna vigintioctopunctata exhibited enhanced insecticidal efficacy compared to E. coli HT115 (DE3). The development of P. chlororaphis BM, a novel dsRNA-expressing bacterium, holds promise for pest management due to its robust dsRNA production and sustained insecticidal activity. This research paves the way for leveraging biocontrol bacteria in RNAi-based pest management strategies. © 2025 Society of Chemical Industry.