The diversity of grassland in upland areas of Czech Republic was studied on selected soil characteristics of these biotopes. In the first phase, 44 soil characteristics were studied and mutual correlations were found between many of them. In the following phase characteristics were chosen which correlated most with other soil characteristics and, at the same time, were easy to evaluate in practise. A great number of correlations were also evidenced between many soil characteristics and the content of humus and nitrogen, which are closely linked to organic matter in soil. In studying these characteristics on selected areas with different types of grassland and consequential cluster analysis and further evaluation, the grassland plots were divided into three groups, from newly established vegetation to species-rich communities. Non-parametric analysis was carried out on the results and a statistically significant difference was proved between the species rich and poorvegetation and carbon and nitrogen content of the soil. Slightly different humus quality (higher amount of HA) was also found under richer vegetation. These results show that at 0-20 cm layer, 58.9 tonnes of carbon ha⁻¹ was measured under species-poor pastureland and 106.1 tonnes of carbon ha under species-rich vegetation. The results showed that besides supporting species diversity, the described quality change can also be important for carbon sequestration. The difference of about 40-50 tonnes of carbon ha and converting 10% of grassland in the Czech Republic to species-rich vegetation would mean sequestration of about 3.9 Mt carbon. If only agroenvironmentally subsidized areas are converted, carbon sequestration in such vegetation could amountto 1.7 Mt.