AbstractSustainable agricultural management practices aimed at improving soil health can alter the soil microbiome, which can influence plant health and defenses against insects. The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major belowground pest of corn. Pest management relies heavily on the planting of transgenic crops expressing Bacillus thuringiensis (Bt) toxins. In this study, we ask how corn-WCR interactions via the soil microbiome are affected by a conservation management system (extended corn-soybean-wheat rotation with cover crops under no-till) compared with a conventional management system (corn-soybean rotation under mulch tillage and no cover crops) when combined with transgenic Bt corn. To do this, we applied soil microbes from the conservation and conventional management systems to two corn lines, one producing Bt and one non-Bt. We then reared Bt-resistant and Bt-susceptible WCR on inoculated seedlings to examine plant and insect changes in fitness. We found that Bt was effective against susceptible larvae in both soil treatments. Bt-resistant larvae were ~ 20% smaller when reared in the presence of soil microbes from the conservation management system. Thus, control of Bt-resistant WCR may be improved in a conservation system without sacrificing Bt effectiveness in susceptible insects. Comparing the microbial communities using 16S rRNA sequencing revealed that management practices influenced the microbiomes associated with the soil and the plant rhizosphere, but not WCR. Our findings suggest value for growers in utilizing conservation management practices, such as no-till and cover crops, in agricultural systems through bottom-up changes to plant–insect interactions via the soil microbiome.