Various extraction methods have been used to determine selenium (Se) concentrations in soils and plants in the second seleniferous regions of China. Our results show tea Se contents in the study area range from 1.009 to 2.6 mg/kg, which reveal that the tea areas in Ziyang County are in seleniferous regions. The four extraction methods evaluated in this study provide different information concerning soil and plant Se levels. The quality control/quality assurance program for this project indicated there is excellent agreement between total soil Se and extractable Se. For example, phosphate extractable Se results from the field investigation and greenhouse study were found to be highly correlated (R 2 > 0.91) by linear regression analyses. Results from rye seedling experiments further show phosphate extractable Se has significant correlations with plant Se uptake and that a 0.1 M solution of KH2PO4 can be used as the extractant of soil available Se. In the acid soil, the Brassica campestris yield could be significantly reduced when the content of Se 6+ –Se ‡ 0.5 mg/kg, and the influence on the yield was not as obvious when the content of Se 6+ –Se reached up to 2.0 mg/kg. The uptake by Brassica campestris of Se 6+ –Se is higher than that of Se 4+ –Se. The main factors influencing the biological availability of soil Se, in order of their importance are CaCO3, the presence of silt grains, organic matter and the presence of clay grains. pH could affect KH2PO4 extractable Se through CaCO3.