<p indent=0mm>Jacalin-related lectins (JRLs) are a group of plant lectins with jacalin protein domain, which play important roles during plant response to biotic and abiotic stress. According their carbohydrate binding specificities, the JRLs were described as two types: galactose-binding acalins (gJRLs) and mannose-binding jacalins (mJRLs). Previous studies have shown that <italic>TaJRL53</italic> encodes a protein with Jacalin and Dirigent domains and can be up-regulated by <italic>Fusarium graminearum</italic> infection. Wheat FHB induced by this pathogen is a disastrous disease, which not only reduces the grain yield, but also contains deoxynivalenol and other toxins, which seriously affect the health of human and livestock. To dissect the gene’s function in FHB resistance, we silenced and overexpressed the <italic>TaJRL53</italic> in wheat respectively. As a result, attenuating <italic>TaJRL53</italic> increased susceptibility to <italic>Fusarium graminearum</italic>. And susceptible wheat transformed with <italic>TaJRL53</italic> over-expression showed higher <italic>Fusarium</italic> head blight resistance, the number of diseased spikelets (NDS) and length of diseased spike rachis (LDR) of transgenics wheat with <italic>TaJRL53</italic> overexpression were significantly reduced than those of wild type. After <italic>Fusarium graminearum</italic> infection, the expression of ROS synthesis pathway, JA synthesis and signaling pathways marker genes, and disease related genes in <italic>TaJRL53</italic> over-expression transgenic plants were higher than those in non-transgenic control. These results suggest that <italic>TaJRL53</italic> enhanced the FHB resistance in wheat through regulating ROS synthesis pathway and JA signal transduction pathways. This study enhanced the understanding of wheat JRLs family genes’ function and laid a foundation for the mechanism dissection of <italic>TaJRL53</italic> resistant to FHB in wheat.