Glycoside Hydrolase Family 16 Enzyme RsEG146 From Rhizoctonia solani AG1 IA Induces Cell Death and Triggers Defence Response in Nicotiana tabacum

Rhizoctonia solani AG1 IA is a harmful necrotrophic fungus responsible for various crop diseases, including maize and rice sheath blight, which can lead to significant production losses. However, the pathogenic mechanisms and the roles of effectors in this pathogen remain poorly understood. In this study, we identified a glycoside hydrolase 16 family gene, RsEG146, from R. solani that was upregulated during its infection of Zea mays leaves. When transiently expressed through agroinfiltration, RsEG146 induced cell death in the leaves of tobacco (Nicotiana tabacum 'Samsun'). The predicted signal peptide of RsEG146 was essential for its cell death-inducing activity, while the conserved enzymic active site was not required. The chitin-binding domain was critical for the cell death-inducing activity of RsEG146, with Gly47 identified as the key residue. Substitution of Gly47 with aspartate, glutamate, or proline significantly impaired the cell death-inducing activity of RsEG146. Additionally, transient and heterogeneous expression of RsEG146 enhanced the pathogenicity of Botrytis cinerea on tobacco, and silencing this gene through spray-induced gene silencing (SIGS) reduced the severity of the disease in maize, indicating that RsEG146 functions as an effector. Furthermore, RsEG146 triggered a plant immune response in tobacco. This study demonstrates that RsEG146 is a potential effector and plays a crucial role in the interactions between R. solani AG1 IA and its host.