Graphene Oxide Induces Toxicity and Alters Energy Metabolism and Gene Expression in Ralstonia solanacearum.

Graphene oxide (GO) is a promising material for development as an antibacterial, phytoprotective agent due to its contact-based antibacterial activity induced by its physical and chemical properties. However, the mechanism underlying the antibacterial effect of GO has yet to be elucidated. In the current study, we investigated the effects of GO on the phytopathogen R. solanacearum at the molecular level with a specific focus on energy metabolism. Under controlled conditions, the bacteriostatic and bactericidal actions of GO were investigated with respect to concentration, treatment time and rotation speed. Transmission electron microscopy (TEM) and destabilization assays revealed that GO caused injury to bacterial cell membrane structures. Furthermore, adenosine triphosphate (ATP) levels decreased after exposure to sheets of GO, while malondialdehyde levels significantly increased, indicating the occurrence of lipid oxidation. A series of genes related to bacterial virulence, motility and oxidative stress were selected to evaluate the molecular mechanism underlying GO’s effects on R. solanacearum. Using quantitative reverse transcription polymerase chain reaction (RT-qPCR), we showed that in the presence of GO, the expression levels of genes involved in virulence and motility were down regulated, with the exception of popA. The phcA, hrpB and flgG genes were significantly downregulated by 2.61-, 3.45- and 4.22-fold, respectively. Conversely, the expression levels of sodB, oxyR and dps, three important oxidative stress genes, were upregulated by 1.82-, 2.17-, and 3.79-fold, respectively. These findings confirmed that cell membrane damage and oxidative stress were responsible for the antibacterial actions of GO, in addition to disturbances to energy metabolism processes.