ABSTRACTGlucose‐regulated protein 78 (Grp78), a crucial molecular chaperone in the endoplasmic reticulum, has been extensively investigated in vertebrates. However, its functional exploration in insects remains limited. This study cloned the full‐length cDNA sequence of Grp78 in Plutella xylostella (L.), which is 2583 bp long. The open reading frame (ORF) is 2004 bp in length and encodes a total of 667 amino acids, including three conserved characteristic sequences of the HSP70 family. PxGrp78 is expressed in various developmental stages of P. xylostella, with the highest expression observed in third instar larvae and higher expression in female adults compared to male adults. The interference with PxGrp78 in female adults was found to significantly reduce the quantity of egg laying and the hatching rate, as well as shorten the oviposition period and down‐regulate the expression of the PxVg gene. These results suggested an important role for PxGrp78 in the reproduction of P. xylostella.