Effects of microplastics and drought on soil ecosystem functions and multifunctionality

Microbes in soils and other environments produce extracellular enzymes to depolymerize and hydrolyze organic macromolecules so that they can be assimilated for energy and nutrients. Measuring soil microbial enzyme activity is crucial in understanding soil ecosystem functional dynamics. The general concept of the fluorescence enzyme assay is that synthetic C-, N-, or P-rich substrates bound with a fluorescent dye are added to soil samples. When intact, the labeled substrates do not fluoresce. Enzyme activity is measured as the increase in fluorescence as the fluorescent dyes are cleaved from their substrates, which allows them to fluoresce. Enzyme measurements can be expressed in units of molarity or activity. To perform this assay, soil slurries are prepared by combining soil with a pH buffer. The pH buffer (typically a 50 mM sodium acetate or 50 mM Tris buffer), is chosen for the buffer's particular acid dissociation constant (pKa) to best match the soil sample pH. The soil slurries are inoculated with a nonlimiting amount of fluorescently labeled (i.e. C-, N-, or P-rich) substrate. Using soil slurries in the assay serves to minimize limitations on enzyme and substrate diffusion. Therefore, this assay controls for differences in substrate limitation, diffusion rates, and soil pH conditions; thus detecting potential enzyme activity rates as a function of the difference in enzyme concentrations (per sample). Fluorescence enzyme assays are typically more sensitive than spectrophotometric (i.e. colorimetric) assays, but can suffer from interference caused by impurities and the instability of many fluorescent compounds when exposed to light; so caution is required when handling fluorescent substrates. Likewise, this method only assesses potential enzyme activities under laboratory conditions when substrates are not limiting. Caution should be used when interpreting the data representing cross-site comparisons with differing temperatures or soil types, as in situ soil type and temperature can influence enzyme kinetics.

Microplastics may enter the soil in a wide range of shapes and polymers. However, little is known about the effects that microplastics of different shapes, polymers, and concentration may have on soil properties and plant performance. To address this, we selected 12 microplastics representing different shapes (fibers, films, foams, and fragments) and polymers, and mixed them each with soil at a concentration of 0.1, 0.2, 0.3, and 0.4%. A phytometer (Daucus carota) grew in each pot during 4 weeks. Shoot, root mass, soil aggregation, and microbial activity were measured. All shapes increased plant biomass. Shoot mass increased by ∼27% with fibers, ∼60% with films, ∼45% with foams, and by ∼54% with fragments, as fibers hold water in the soil for longer, films decrease soil bulk density, and foams and fragments can increase soil aeration and macroporosity, which overall promote plant performance. By contrast, all shapes decreased soil aggregation by ∼25% as microplastics may introduce fracture points into aggregates and due to potential negative effects on soil biota. The latter may also explain the decrease in microbial activity with, for example, polyethylene films. Our findings show that shape, polymer type, and concentration are key properties when studying microplastic effects on terrestrial systems.