Characteristics of resistance of VC6089A, a mungbean, Vigna radiata (L.) Wilczek, bred by using a wild Vigna species, V. sublobata (Roxburgh) Verdcourt (accession no. TC1966), and containing a novel protein, VrD1, were investigated against the cowpea weevil, Callosobruchus maculatus (F.). The seeds of VC6089A showed high level of resistance; >96% of the bruchid eggs failed to develop into adults, whereas 85% of eggs laid on susceptible cultivar VC1973A became adults. Mortality of surviving bruchids raised for five generations on VC6089A remained higher than 96%; however, female adults maintained high fecundity and thus showed a positive population growth through these generations. We therefore cannot exclude the possibility that the beetles could develop resistance to the resistant mungbean VC6089A. The protein VrD1 purified from seeds of VC6089A showed marked toxicity to C. maculatus when beetles were reared on artificial seeds containing varying levels of VrD1. Thorough inhibition of development was observed when artificial seeds containing 0.2% (wt:wt) VrD1 was provided for insect feeding. Our findings demonstrated the insecticidal activity of VC6089A mungbean seeds and VrD1 protein against C. maculatus. These results may facilitate safer control against bruchid infestation.