The effect of organic, mineral and combined organic and mineral fertilization of soils on the winter wheat yields and nutrient contents in soils was evaluated in long-term field experiments. Two sites with different soil characteristics were evaluated -Lukavec u Pacova (cambisol) and Ivanovice na Hané (degraded chernozem). The type of fertilization influenced wheat yields. Nutrient uptake by winter wheat was higher under nitrogen (N) fertilization, which resulted in a negative balance of phosphorus (P) and potassium (K) and to a decrease of nutrient contents in the more fertile soils at Ivanovice na Hané. Two soil tests (Mehlich 3 method and NH 4 -acetate method) were used to determine P and K availability. The mineral nitrogen fertilization negatively and significantly affected NH 4 -acetate extractable concentrations of nutrients in the soils and these were lower in comparison with concentrations of P and K determined by Mehlich 3 method. Relative availability of P in alkaline soils from Ivanovice treated with mineral N increased while the soil pH decreased.