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<jats:p> Ziadi, N., Angers, D. A., Gagnon, B., Lalande, R., Morel, C., Rochette, P. and Chantigny, M. H. 2014. Long-term tillage and synthetic fertilization affect soil functioning and crop yields in a corn–soybean rotation in eastern Canada. Can. J. Soil Sci. 94: 365–376. Adoption of conservation practices can induce beneficial changes to soil properties and related crop yields in which magnitude varies according to soil and climatic conditions but usually increases with time. A long-term field experiment was initiated in 1992 at L'Acadie in southern Quebec on a clay loam soil to evaluate the effect of tillage [mouldboard plow (MP) vs. conservation (CT)], synthetic N fertilization (0, 80, and 160 kg N ha<jats:sup>−1</jats:sup>) and synthetic P fertilization (0, 17.5, and 35 kg P ha<jats:sup>−1</jats:sup>) on soil functioning and grain yields of a corn–soybean rotation. Soil tillage was performed every year while synthetic fertilizers were applied only to the corn. Results obtained 12 to 20 yr after initiation of the study indicated that CT enhanced organic C accumulation, NO<jats:sub>3</jats:sub>-N, P and K availability, microbial biomass and activity, and microbial community structure in the upper soil layer, likely due to leaving crop residues at the soil surface. The MP practice resulted in greater organic C content deeper, near the bottom of the plow layer, which promoted soil microbial activity at that depth. However, soil N<jats:sub>2</jats:sub>O emissions were not affected by tillage. The N and P fertilization increased the availability of these nutrients, but had no significant effect on the soil microbial biomass, activity, and structure. Linear relationships were established between soil available P and cumulative P budgets obtained under MP or 0 kg P ha<jats:sup>−1</jats:sup> under CT. Crop yields varied by year in this study but on average, MP yielded 10% more corn and 13% more soybeans than CT. Corn yield increased linearly with added synthetic N each year, whereas soybean yield was little affected by residual N, and both crops did not respond to fertilizer P. Response to N fertilization did not differ due to tillage or P. Despite higher costs associated with plowing, the profitability of MP was greater than CT on this clay loam soil due to greater yields. Specialized management practices (e.g., delayed planting, better herbicide selection, fall cover crop, in-row tillage) might help to improve CT performance on these cool, humid fine-textured soils. </jats:p>