Activation of Early Defense Signals in Seedlings of Nicotiana benthamiana Treated with Chitin Nanoparticles

Chitin is an excellent material for the synthesis of nanoparticles because it is an elicitor and can form nanostructured materials. The application of chitin nanoparticles (CNPs) in plants can activate early defense responses associated with chitin. In this study, CNPs were synthesized by water in oil (W/O) emulsion using an aqueous chitin solution. The CNPs were characterized and used to evaluate the activation of genes related to early responses to chitin and the production of reactive oxygen species (ROS) on seedlings of Nicotiana benthamiana. The CNPs had an average size of 280 nm in diameter, a polydispersity of 0.299, a surface charge of 26.9 mV, and their chemical composition was corroborated by the disappearance of microaggregated CNPs treated with chitinases observed under a microscope. Seedlings treated with CNPs for one hour revealed increments in the expression of genes STZ, ATL2, and MAPK3, in contrast when they were treated with chitin oligomers, and no changes in gene CERK1 was detected in both conditions. Finally, the synthesis of ROS mediated by CNPs was detected in seedlings, which was higher than those generated by the treatment of chitin oligomers. These results demonstrated the capability to generate CNPs by emulsion, which are capable of triggering responses related to early defense in N. benthamiana more efficiently than chitin oligomers.