Osmia cornifrons plays a major role in the pollination of orchards, but basic information on vitellogenin and the oocyte development is limited. To better understand vitellogenin in hymenopteran insects, we cloned a cDNA encoding vitellogenin from the hornfaced bee O. cornifrons. O. cornifrons vitellogenin cDNA contains 5477 bp with an open reading frame of 1,783 amino acid residues, and has a predicted molecular mass of approximately 200.21 kDa and a pI of 6.55. O. cornifrons vitellogenin possesses four consensus (RXXR/S) cleavage sites and has conserved DGXR and GL/ICG motifs in the C-terminus. The deduced amino acid sequence of the O. cornifrons vitellogenin cDNA showed a 66% identity with Megachile rotundata, 53% to Apis mellifera, 51% to Bombus ignitus, and 42%-30% with other hymenopteran insect vitellogenins. Phylogenetic analysis showed that O. cornifrons vitellogenin clustered with vitellogenins from Megachildae, Apidae, Vespidae, and Formicidae species but not with those from Pteromalidae, Aphelinidae or Ichneumonidae species.