Bradford reactive soil protein in Appalachian soils: distribution and response to incubation, extraction reagent and tannins

Bradford reactive soil protein (BRSP) is thought to correlate to glomalin, an important soil glycoprotein that promotes soil aggregate formation and may represent a significant pool of stable soil organic matter (SOM). However, more information is needed about its importance in Appalachian soils and its relationships with other soil properties. We measured BRSP in 0-20 cm soil from pastures, hayfields, cultivated fields or forest areas in southern West Virginia. Highest amounts of BRSP were found near the soil surface and decreased significantly with depth for all land uses except cultivated sites. Forest and pasture sites contained more BRSP than hayfields or cultivated fields but these differences occurred only in the 0-5 cm depth. Overall averages of C and N in BRSP represented about 4.0 and 6.5% of the total soil C and N respectively. During a 395 day soil incubation, we found CO2-C evolution rates comparable to other studies but only small changes in BRSP (< 10%) including some evidence for increases during incubation. Sodium citrate, sodium pyrophosphate, and sodium oxalate recovered significantly more BRSP from soil than the other extractants we tested with highest extraction efficiencies observed for sodium citrate and pyrophosphate. Recovery of BRSP appears related to negative charge and buffering capacity of both the soil and extractant. Extractants with low negative charge had little buffering capacity and yielded little BRSP. Tannic acid appeared to increase extraction of BRSP but less soluble-N was recovered from tannin-treated samples than from untreated controls and E4/E6, the ratio of absorbance at 465 and 665 nm, decreased, evidence for the formation of larger or heavier molecules. Formation of dark-colored substances during extraction suggests the colorimetric Bradford assay may overestimate soil protein when tannins are present. Recovery of less soluble-N from soil extracts and lower E4/E6 ratios suggests tannins may bind with soil constituents themselves or form non-extractable N-containing complexes.