The long-term, poly-factorial, field experiment on Typical chernozemon the Balti steppe (Republic of Moldova) demonstrates the benefits of crop rotation that includes a perennial legume-grass mixture: higher crop yields; better accumulation of soil organic matter and nitrates, and uptake of soil phosphate; and more efficient use of soil water, especially on unploughed plots. The agronomic effect of soil tillage is determined by the interaction with crop sequence and fertilization in the crop rotation. The capacity of the soil to accumulate soil water and consumption of soil water are higher in crop rotation with perennial legumes and grasses, especially under the plough. Crop rotation with the mixture of perennial legume and grasses under ploughless tillage increases the accumulation of soil organic matter relative to other variants. By using a crop rotation with a mixture of perennial legume and grasses, farmers have real possibilities to decrease the dependence on industrial inputs.