Viruses rely on host cellular metabolism for energy and macromolecule synthesis during replication. Therefore, there is a close relationship between viral replication and cellular metabolism. rhabdovirus (SCRV) causes significant economic losses in the Chinese perch () industry. Thus, it is important to address the interaction between SCRV replication and host cells to prevent and control SCRV. However, little is known about the relationship between SCRV replication and cellular metabolism. In addition, glutamine is an abundant amino acid necessary for energy generation and macromolecule synthesis in cells. Glutamine is also essential for the infection and replication of some viruses. Glutaminolysis and reductive glutamine metabolism (RGM) are the main pathways of glutamine metabolism. Particularly, the RGM pathway plays an important role in tumor cells. However, the metabolic pathway of glutamine involved in viral replication remains largely uncharacterized. To clarify the interaction between SCRV replication and RGM, we analyzed the expression of key enzymes in the RGM pathway in Chinese perch brain (CPB) cells following infection with SCRV, using quantitative real-time PCR (qRT-PCR) and western blotting. Our results showed that SCRV replication was decreased in CPB cells cultured in a glutamine-depleted medium, indicating that glutamine is required for efficient SCRV replication. In addition, SCRV infection promoted the expression of key enzymes involved in the RGM pathway in CPB cells, particularly the expression of isocitrate dehydrogenase 2 (IDH2), indicating that RGM in CPB cells is altered during SCRV infection. SCRV proliferation was also inhibited in CPB cells cultured in the presence of bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl)ethyl sulfide (a glutaminase inhibitor), (-)-epigallocatechin monogallate (a glutamate dehydrogenase inhibitor), 5-(tetradecyloxy)-2-furoic acid (an acetyl-CoA carboxylase inhibitor), or 4-methylene-2-octyl-5-oxotetrahydrofuran-3-carboxylic acid (a fatty acid synthase inhibitor), suggesting that the RGM pathway is beneficial for SCRV replication and proliferation. Furthermore, downregulation of the IDH2 gene, the product of which converts α-ketoglutarate (α-KG) to citrate, significantly inhibited the yield of SCRV and the expression of the SCRV structural protein. Conversely, overexpression of the IDH2 gene significantly increased the yield of SCRV and the expression of SCRV structural protein, indicating that IDH2 plays an important role in SCRV replication and proliferation. These results indicated that SCRV infection induces RGM to accommodate the biosynthetic and energy needs required for its efficient replication and proliferation. These findings provide new insights into viral pathogenesis and antiviral treatment strategies.