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The soybean aphid ( Aphis glycines ) continues to threaten soybean production in the United States. A suite of management strategies, such as planting aphid-resistant cultivars, has been successful in controlling soybean aphids. Several  Rag  genes (resistance against  A. glycines ) have been identified, and two are currently being deployed in commercial soybean cultivars. However, the mechanisms underlying  Rag -mediated resistance are yet to be identified. In this study, we sought to determine the nature of resistance conferred by the  Rag5  gene using behavioral, molecular biology, physiological, and biochemical approaches. We confirmed previous findings that plants carrying the  Rag5  gene were resistant to soybean aphids in whole plant assays, and this resistance was absent in detached leaf assays. Analysis of aphid feeding behaviors using the electrical penetration graph technique on whole plants and detached leaves did not reveal differences between the  Rag5  plants and Williams 82, a susceptible cultivar. In reciprocal grafting experiments, aphid populations were lower in the  Rag5/rag5  (Scion/Root stock) chimera, suggesting that  Rag5- mediated resistance is derived from the shoots. Further evidence for the role of stems comes from poor aphid performance in detached stem plus leaf assays. Gene expression analysis revealed that biosynthesis of the isoflavone kaempferol is upregulated in both leaves and stems in resistant  Rag5  plants. Moreover, supplementing with kaempferol restored resistance in detached stems of plants carrying  Rag5 . This study demonstrates for the first time that  Rag5 -mediated resistance against soybean aphids is likely derived from stems.

The Source of Rag5-Mediated Resistance to Soybean Aphids Is Located in the Stem