The Rag1 gene confers antibiotic resistance to soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), and in 2010, varieties expressing Rag1 were released for commercial use in the United States. We do not know how Rag1 varieties will influence the broader community of defoliating insects that inhabit soybean fields. In 2010 and 2011, the preference and performance of pest insects that defoliate soybeans [Glycines max (L.) Merr] were tested using Rag1 and aphidsusceptible varieties. Three coleopterans and four lepidopterans were used: northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae); southern corn rootworm, Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae); bean leaf beetle, Ceratoma trifurcata Förster (Coleoptera: Chrysomelidae); fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae); corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae); soybean looper, Chrysodeix includens (Walker) (Lepidoptera: Noctuidae); and velvetbean caterpillar, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae). The preference of insects was evaluated in choice and no-choice tests using Rag1 and susceptible soybeans. Lepidopterans also were evaluated on Rag1 leaves using four nutritional indices: relative growth rate, approximate digestibility, and efficiency of conversion of ingested material. In the majority of preference tests, no effect of Rag1 was detected, and in cases where preferences were found, there was no consistent pattern of preference for Rag1 vs. susceptible leaf tissue. Helicoverpa zea demonstrated a preference for resistant leaf tissue, but this was dependent on the genetic background of the variety. Evaluations of nutritional indices indicated that three species of Lepidoptera, S. frugiperda, H. zea, and A. gemmatalis, displayed reduced conversion efficiency for Rag1 soybeans, suggesting effects of antibiosis.