Cover crop mixture diversity, biomass productivity, weed suppression, and stability.

The diversity-productivity, diversity-invasibility, and diversity-stability hypotheses propose that increasing species diversity should lead, respectively, to increased average biomass productivity, invasion resistance, and stability. We tested these three hypotheses in the context of cover crop mixtures, evaluating the effects of increasing cover crop mixture diversity on aboveground biomass, weed suppression, and biomass stability. Twenty to forty cover crop treatments were replicated three or four times at eleven sites using eighteen species representing three cover crop species each from six pre-defined functional groups: cool-season grasses, cool-season legumes, cool-season brassicas, warm-season grasses, warm-season legumes, and warm-season broadleaves. Each species was seeded as a pure stand, and the most diverse treatment contained all eighteen species. Remaining treatments included treatments representing intermediate levels of cover crop species and functional richness and a no cover crop control. Cover crop seeding dates ranged from late July to late September with both cover crop and weed aboveground biomass being sampled prior to winterkill. Stability was assessed by evaluating the variability in cover crop biomass for each treatment across plots within each site. While increasing cover crop mixture diversity was associated with increased average aboveground biomass, we assert that this was the result of the average biomass of the pure stands being drawn down by low biomass species rather than due to niche complementarity or increased resource use efficiency. At no site did the highest biomass mixture produce more than the highest biomass pure stand. Furthermore, while increases in cover crop mixture diversity were correlated with increases in weed suppression and biomass stability, we argue that this was largely the result of diversity co-varying with aboveground biomass, and that differences in aboveground biomass rather than differences in diversity drove the differences observed in weed suppression and stability.

Soil electrical conductivity and water content affect nitrous oxide and carbon dioxide emissions in intensively managed soils.

Accumulation of soluble salts resulting from fertilizer N may affect microbial production of N(2)O and CO(2) in soils. This study was conducted to determine the effects of electrical conductivity (EC) and water content on N(2)O and CO(2) production in five soils under intensive cropping. Surface soils from maize fields were washed, repacked and brought to 60% or 90% water-filled pore space (WFPS). Salt mixtures were added to achieve an initial in situ soil EC of 0.5, 1.0, 1.5 and 2.0 dS m(-1). The soil cores were incubated at 25 degrees C for 10 d. Average CO(2) production decreased with increasing EC at both soil water contents, indicating a general reduction in microbial respiration with increasing EC. Average cumulative N(2)O production at 60% WFPS decreased from 2.0 mg N(2)O-N m(-2) at an initial EC of 0.5 dS m(-1) to 0.86 mg N(2)O-N m(-2) at 2.0 dS m(-1). At 90% WFPS, N(2)O production was 2 to 40 times greater than that at 60% WFPS and maximum N(2)O losses occurred at the highest EC level of 2.0 dS m(-1). Differences in the magnitude of gas emissions at varying WFPS were due to available substrate N and the predominance of nitrification under aerobic conditions (60% WFPS) and denitrification when oxygen was limited (90% WFPS). Differences in gas emissions at varying soil EC may be due to changes in mechanisms of adjustment to salt stress and ion toxicities by microbial communities. Direct effects of EC on microbial respiration and N(2)O emissions need to be accounted for in ecosystems models for predicting soil greenhouse gas emissions.

Rapid spectrophotometric determination of 2,4,6-trinitrotoluene in a Pseudomonas enzyme assay.

Although TNT (2,4,6-trinitrotoluene) and its degradation products can be quantified by HPLC, this method is not suitable for simultaneous analyses of the numerous samples typically encountered in enzyme studies. To solve this problem, we developed a simple and rapid spectrophotometric assay for TNT and tested the procedure using partially purified nitroreductase(s) from a Pseudomonas aeruginosa isolate, which transformed TNT in the culture medium. In highly alkaline solution, TNT (pK(a)=11.99) exhibits significant absorbance at 447 nm, while major metabolites, 2-amino-4, 6-dinitrotoluene (2ADNT), 4-amino-2,6-dinitrotoluene (4ADNT), and 2,6-diamino-4-nitrotoluene (2,6DANT) display no absorbance at this wavelength. Assay mixtures of TNT, Tris-HCl buffer, a reductant, and the enzyme(s) were analyzed by measuring absorbance 4 min after adjusting the pH to 12.2. TNT transformation to colorless metabolites was linear with respect to protein and substrate concentrations. Using the assay, we determined that TNT nitroreductase(s) from the isolate required an electron donor and preferred NADH to NADPH. TNT transformation increased when NAD was recycled to NADH using glucose-6-phosphate (GP) and glucose-6-phosphate dehydrogenase (GPDH). Enzymatic transformation of TNT was completely inhibited by Cu(2+) (5 mM) and was partially inhibited by other divalent metallic cations. Because the assay is sensitive to ammonium sulfate, dithiothreitol, ascorbic acid, and sodium phosphate, extracts should be assayed in the absence of these components.

Denitration of 2,4,6-trinitrotoluene by Pseudomonas savastanoi.

Past disposal of wastewaters containing 2,4,6-trinitrotoluene (TNT) at the former Nebraska Ordnance Plant has resulted in numerous acres of TNT-contaminated soil. Examining the microbial population of these soils revealed several TNT-tolerant Pseudomonas spp. We selected one species, P. savastanoi, to determine its ability to transform TNT. Pure culture experiments were performed in pseudomonas minimal medium containing 0.31 mM TNT (70 mg TNT . L(-1)) under varied nutrient and cell density regimes. Experiments with TNT as a sole C or N source showed that P. savastanoi has the ability to denitrate TNT, as evidenced by production of 2,4-dinitrotoluene (2,4-DNT) and NO2- with time. TNT denitration and formation of 2,4-DNT were enhanced by removing NH4+ and adding NO2- to the growth medium. In all experiments, 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT) appeared as incidental reduction products. Glucose addition to the medium enhanced 2-ADNT and 4-ADNT production and decreased denitration of TNT. Mid-log phase cells rapidly transformed [ring-14C(U)]TNT but were unable to mineralize significant quantities of TNT, as evidenced by conversion of less than 1% of the label to 14CO2. These results indicate that P. savastanoi is a TNT-tolerant pseudomonad that can promote TNT degradation through reductive denitration and nitro moiety reduction.