Most agricultural products produced in naturally seleniferous areas have problems including large variation and instability of Se content. Therefore, it is of great significance to establish a suitable extraction method for the development and utilization of seleniferous soil resources. In this article, 40 soil samples were collected from the naturally seleniferous areas in Jianshi County, Enshi Tujia and Miao Autonomous Prefecture, Hubei Province and used for rice pot experiment to establish a method for extracting effective selenium from acidic and seleniferous soil. The sequential extraction method and 10 commonly used single step extraction methods were used to extract and determine the forms of selenium and available selenium in soil. The correlation between the content of effective selenium extracted by different extraction methods and the selenium absorbed by rice plants was analyzed. The results showed that the content of water-soluble and exchangeable selenium in the acidic and seleniferous soil in Jianshi was relatively low, ranging from 0.12 to 46.0 μg/kg and from 0.58 to 197.35 μg/kg, respectively. Selenium mainly existed in forms of organic bound and residual selenium that are difficult for plants to utilize. The content of selenium in iron manganese oxide bound state, organic bound state, and residual state was 0.01-0.34 mg/kg, 0.05-0.88 mg/kg, and 0.04-2.72 mg/kg, respectively. Among dozens of single step extractants, 0.5 mol/L NaHCO₃ and 0.1 mol/L KH₂PO₄ solutions had higher extraction efficiency for available selenium in soil. Among different forms of selenium in soil, the correlation between exchangeable selenium and the content of selenium in plant was the highest (r=0.80, P<0.01), while the correlation between other forms of selenium and the content of selenium in plant was lower than that of exchangeable selenium. Among the single step extraction methods, the content of available selenium in soil extracted with 0.1 mol/L KH₂PO₄ solution and 0.5 mol/L NaHCO₃ solution was higher, but the correlation between the former and the content of selenium in plant was more significant (r=0.83, P<0.01). It is indicated that the extraction method with 0.1 mol/L KH₂PO₄ is the most suitable method for extracting biologically available selenium from weakly acidic and seleniferous soil if considering the extraction efficiency and its correlation with the content of selenium in plant.