Background and Aims. The study aims to explore the impact of advancements in wheat genetics on root structure and rhizosphere biology, which are still not fully understood. Specifically, we investigated various factors including the exudation of carboxylates, colonization by arbuscular mycorrhizal fungi, microbial activity, and root architecture in winter wheat varieties that have been released between 1965 and 2020. Methods. To conduct our study, we sowed fourteen different winter wheat varieties with four replicates on acidic Andisol at field conditions. Complete root systems and soil samples were extracted using a tractor-mounted hydraulic sampler tube of 3.5 cm diameter, which reached a depth of 60 cm. Results. In this sense, succinate showed a significant increase by 21%. Mycorrhizal colonization was inversely proportional to P concentrations and all varieties showed higher microbial activity at anthesis. The longest roots were found in varieties released after the year 2000, but no significant differences were found in other root architecture parameters. There was no clear pattern observed in root architecture or biological activity as a function of the year of release. Plant genetics moderated root architecture, carboxylate exudation, microbial activity, and mycorrhizal colonization, all of which are affected by high P concentrations. Conclusions. This study investigated plant-microorganism interactions, often overlooked due to root system analysis challenges. Older wheat varieties showed higher carboxylate exudation. We identified wheat varieties with potential for improved root systems and crop efficiency.