Identification of Insect Nephotettix cincticeps Proteins Interacting With Rice Dwarf Virus‐Encoding Pns10 Tubule by DUALmembrane Yeast Two‐Hybrid System

Rice dwarf virus (RDV) exploits virus-induced non-structural proteins Pns10 tubules to overcome membrane or tissue barriers for viral spread within its insect vectors, Nephotettix cincticeps. However, whether vector factors are involved in the motility of virus-containing tubules and, if so, which ones, remains largely unknown. In this study, a high-throughput DUALmembrane yeast two-hybrid system was employed to screen for N. cincticeps proteins interacting with RDV Pns10 tubule. Our results demonstrate that RDV Pns10 interacts with 64 candidate proteins from the vector, which were classified into nine molecular function categories. Furthermore, we confirmed that the expression levels of 14 selected candidate genes, primarily including three energy-related mitochondria genes, three cytoskeleton-related genes, three ribosomal protein synthesis genes, two stress-related DnaJ protein genes, and three intracellular transport-related proteins, were differently modulated upon interaction with RDV Pns10 protein. In conclusion, this study provides new insights into the role and importance of RDV Pns10 interaction with vector proteins during viral infection. It also enhances our understanding of the biochemistry and physiological responses of N. cincticeps to viral infection.