A method for linking in situ activities of hydrolytic enzymes to associated organisms in forest soils

A root window-based, enzyme-imprinted, membrane system has been modified to enable visualization of the activities of hydrolytic enzymes (acid phosphatase, aminopeptidase, chitinase, and β-glucosidase) in situ in forest soils. The approach can be used to correlate the distribution of enzyme activity with visible features such as roots, mycorrhizas, or mycelial mats. In addition, it enables accurate spatial soil sampling for analysis of microbial communities associated with enzyme activities. The substrates are colorimetric conjugates of napthol, where color develops instantly in the field, or fluorimetric conjugates of 4-methylumbelliferone, whose fluorescent products are detected by a gel-documenting system. The method will allow important questions about the relationship between taxonomic and functional diversity of soil microorganisms to be addressed and identification of enzyme activity hot-spots in soil.