Expression of the ripAA Gene in the Soilborne Pseudomonas mosselii Can Promote the Control Efficacy against Tobacco Bacterial Wilt

SIMPLE SUMMARY: Tobacco bacterial wilt caused by Ralstonia solanacearum brings large economic losses every year. Currently, an increasing number of biocontrol agents have been widely used in the control of this disease, but they cannot replace chemical agents, mostly due to the poorer control effect. Therefore, in this study, the avirulence gene ripAA from Ralstonia solanacearum, which determines incompatible interactions with tobacco plants, was introduced into the biocontrol agent Pseudomonas mosselii to increase the efficacy against Ralstonia solanacearum. The newly engineered strain can improve the systemic resistance and elicit a primary immune response of plants. Our research not only provides a new strategy for the genetic modification of biocontrol agents, in which a number of avirulence genes from the pathogen or plant can be tested to be expressed in different biocontrol agents to antagonize this plant disease, but also helps the study of the interaction between the phytopathogenic avirulence gene and the host. ABSTRACT: The environmental bacterium Pseudomonas mosselii produces antagonistic secondary metabolites with inhibitory effects on multiple plant pathogens, including Ralstonia solanacearum, the causal agent of bacterial wilt. In this study, an engineered P. mosselii strain was generated to express R. solanacearum ripAA, which determines the incompatible interactions with tobacco plants. The ripAA gene, together with its native promoter, was integrated into the P. mosselii chromosome. The resulting strain showed no difference in antimicrobial activity against R. solanacearum. Promoter-LacZ fusion and RT-PCR experiments demonstrated that the ripAA gene was transcribed in culture media. Compared with that of the wild type, the engineered strain reduced the disease index by 9.1% for bacterial wilt on tobacco plants. A transcriptome analysis was performed to identify differentially expressed genes in tobacco plants, and the results revealed that ethylene- and jasmonate-dependent defense signaling pathways were induced. These data demonstrates that the engineered P. mosselii expressing ripAA can improve biological control against tobacco bacterial wilt by the activation of host defense responses.