G. W. McCarty

Effect of freeze-thaw events on mineralization of soil nitrogen

SummaryIn humid regions of the United States there is considerable interest in the use of late spring (April–June) soil NO3−concentrations to estimate fertilizer N requirements. However, little information is available on the environmental factors that influence soil NO3−concentrations in late winter/early spring. The influence of freeze-thaw treatments on N mineralization was studied on several central Iowa soils. The soils were subjected to temperatures of-20°C or 5°C for 1 week followed by 0–20 days of incubation at various temperatures. The release of soluble ninhydrin-reactive N, the N mineralization rate, and net N mineralization (mineral N flush) were observed. The freeze-thaw treatment resulted in a significant increase in the N mineralization rate and mineral N flush. The N mineralization rate in the freeze-thaw treated soils remained higher than in non-frozen soils for 3–6 days when thawed soils were incubated at 25°C and for up to 20 days in thawed soils incubated at 5°C. The freeze-thaw treatments resulted in a significant release of ninhydrin-reactive N. These values were closely correlated with the mineral N flush (r2=0.84). The release of ninhydrin-reactive N was more closely correlated with biomass N (r2=0.80) than total N (r2=0.65). Our results suggest that freeze-thaw events in soil disrupt microbial tissues in a similar way to drying and re-wetting or chloroform fumigation. Thus the level of mineral N released was directly related to the soil microbial biomass. We conclude that net N mineralization following a spring thaw may provide a significant portion of the total NO3−present in the soil profile.

Denitrification capacity and denitrification potential of subsurface soils

Abstract Although numerous studies of denitrification in surface soils have been reported, few attempts have been made to study denitrifying activity in subsurface soils. We collected samples of four Iowa soil profiles to a depth of 3 m and measured their population of denitrifying bacteria and their capacity and potential for denitrification. Their denitrification capacity was assessed from their ability to reduce nitrate when incubated anaerobically (helium atmosphere) at 30°C for 72 hours after treatment with nitrate, and their denitrification potential was assessed from their corresponding ability when incubated anaerobically after treatment with both nitrate and organic carbon (as glucose). We found that the denitrification potentials of the subsurface samples studied greatly exceeded their denitrification capacities and that whereas both the population of denitrifying bacteria and the denitrification capacity of the samples decreased appreciably with depth in the profile, the denitrification potenti...

Availability of organic carbon for denitrification of nitrate in subsoils

SummaryPrevious work in our laboratory indicated that the slow rate of denitrification in Iowa subsoils is not due to a lack of denitrifying microorganisms, but to a lack of organic C that can be utilized by these microorganisms for reduction of NO3−. This conclusion was supported by studies showing that drainage water from tile drains under agricultural research plots contained only trace amounts of organic C and had very little, if any, effect on denitrification in subsoils. Aqueous extracts of surface soils promoted denitrification when added to subsoils, and their ability to do so increased with increase in their organic C content. Amendment of surface soils with corn and soybean residues initially led to a marked increase in the amounts of organic C in aqueous extracts of these soils and in the ability of these extracts to promote denitrification in subsoils, but these effects were short-lived and could not be detected after incubation of residue-treated soils for a few days. We conclude from these observations that water-soluble organic C derived from plant residues is decomposed so rapidly in surface soils that very little of this C is leached into subsoils, and that this largely accounts for the slow rate of denitrification of nitrate in subsoils.

Regulation of urease production in soil by microbial assimilation of nitrogen

SummaryStudies of the effects of different forms of N on urease production in soils amended with organic C showed that although microbial activity, as measured by CO2 production, was stimulated by the addition of NH4+ or NO3- to C-amended soils (200 μmol glucose-C g−1 soil), urease production was repressed by these forms of N. The addition of L-methionine sulfoximine, an inhibitor of inorganic N assimilation by microorganisms, relieved the NH4+ and NO3- repression of urease production in C-amended soil. The addition of sodium chlorate, an inhibitor of NO3- reduction to NH4+ by microorganisms, relieved the NO3- repression of urease production, but did not eliminate the repression associated with NH4+. These observations indicate that microbial production of urease in C-amended soils is not directly repressed by NH4+ or NO3-, but by products formed by microbial assimilation of these forms of N. This conclusion is supported by our finding that the biologically active L-isomers of alanine, arginine, asparagine, aspartate, and glutamine, repressed urease production in C-amended soil, whereas the D-isomers of these amino acids had little or no influence on urease production. This work suggests that urease synthesis by soil microorganisms is controlled by the global N regulon.

Laboratory evaluation of dicyandiamide as a soil nitrification inhibitor

Abstract Laboratory studies to evaluate dicyandiamide (DCD) as a soil nitrification inhibitor showed that it is considerably more effective than several compounds that have been patented or proposed as fertilizer amendments for retarding nitrification of fertilizer nitrogen (N) in soil, but is considerably less effective than 2‐ethynylpyridine, nitrapyrin (N‐Serve), etridiazole (Dwell), 3‐methylpyrazole‐l‐carboxamide (MPC), or 4‐amino‐l,2,4‐triazole (ATC). Other findings in studies reported were as follows: a) DCD is more effective for inhibiting nitrification of ammonium‐N than of urea‐N; b) the effectiveness of DCD as a nitrification inhibitor is markedly affected by soil temperature and soil type and is limited by the susceptibility of DCD to leaching; c) DCD has very little, if any, effect on urea hydrolysis, denitrification, or seed germination in soil; d) products of DCD decomposition in soil (guanylurea and guanidine) have little, if any, effect on nitrification compared with DCD; e) in the absence...

Effect of N-(n-butyl) thiophosphoric triamide on hydrolysis of urea by plant, microbial, and soil urease

SummaryComparison of the effects of N-(n-butyl) thiophosphoric triamide (NBPT) and phenylphosphorodiamidate (PPD) on hydrolysis of urea by plant (jackbean), microbial (Bacillus pasteurii), and soil urease showed that whereas NBPT was considerably more effective than PPD for inhibiting hydrolysis of urea added to soil, it was much less effective than PPD for inhibiting hydrolysis of urea by plant or microbial urease. Studies to account for this observation indicated that NBPT is rapidly decomposed in soil to a compound that is much more effective than NBPT for inhibition of urease activity and that this compound is N-(n-butyl) phosphoric triamide.

Inhibition of nitrification in soil by heterocyclic nitrogen compounds

SummaryThe relationship between the structures of diverse heterocyclic nitrogen (N) compounds and the effectiveness of these compounds for the inhibition of nitrification in soil was studied by determining the effects of different amounts of 12 unsubstituted and 33 substituted heterocyclic N compounds on the production of (NO2−+NO3−)-N in soils incubated at 25 °C for 21 days after treatment with ammonium sulfate. The results showed that unsubstituted heterocyclic N compounds containing two adjacent ring N atoms inhibit nitrification in soil and that two of these compounds, pyrazole and 1,2,4-triazole, are potent inhibitors. They also showed that several substituted pyrazoles and thiadiazoles are good inhibitors of nitrification in soil (e.g., 3-methylpyrazole and 3,4-dichloro-1,2,5-thiadiazole).

Factors affecting the availability of organic carbon for denitrification of nitrate in subsoils

SummaryRecent work in our laboratory indicated that the slow rate of denitrification in Iowa subsoils is not due to a lack of denitrifying microorganisms, but to a lack of organic C that can be utilized by these microorganisms for reduction of nitrate. To identify factors affecting the availability of leachable organic C in surface soils capable of promoting denitrification in subsoils, we studied the effects of freezing and drying and of plants and plant residues on the amounts of water-soluble organic C in surface soils and the ability of this organic C to promote denitrification in subsoils. We found that aqueous extracts of field-moist, frozen, and air-dried surface soils promoted denitrification in subsoils and that their stimulatory effects on denitrification were highly correlated (r=0.93) with their organic C contents and decreased in the order air-dried soils ≫ frozen soils >field-moist soils. But a detailed study of the effect of drying a surface soil to different water tensions indicated that drying of soils under natural conditions is not likely to lead to a substantial increase in their content of water-soluble organic C. Amendment of surface soils with corn or soybean residues led to a marked increase in the amount of organic C in aqueous extracts of the soils and in the ability of these extracts to promote denitrification in subsoils. These effects of plant residues could not be detected after incubation of residue-treated soils for a few days under aerobic conditions, but they increased markedly with an increase in the time of incubation from 1 to 10 days when residue-treated soils were incubated under anaerobic conditions. Analyses for organic acids indicated that this increase was largely due to fermentative production of acetic, propionic, and butyric acids by soil microorganisms. Growth chamber studies showed that growth of corn, soybean, wheat, and sorghum plants on surface soil did not significantly increase the organic C content of leachates of the soil or the ability of these leachates to promote denitrification in subsois. We conclude that plant residues are a major source of the leachable organic C in surface soils that is capable of promoting denitrification in subsoils.

Inhibition of plant and microbial ureases by phosphoroamides

Enzyme kinetic studies of inhibition of plant (jackbean) and microbial (Bacillus pasteurii) ureases by eight phosphoroamides [phenylphosphorodiamidate, 4-chlorophenylphosphorodiamidate, phosphoric triamide, N-(diaminophosphinyl)benzamide, N-(diaminophosphinyl)benzeneacetamide, 4-chloro-N-(diaminophosphinyl)benzamide, N-(4-nitrophenyl)phosphoric triamide, N-(diaminophosphinyl)-3-pyridinecarboxamide] demonstrated that these compounds are slow, tight-binding inhibitors of urease enzymes. Measurement of the dissociation constants (Ki*) of the enzyme-inhibitor complexes (E · I*) formed by interaction of the ureases and phosphoroamide inhibitors studied showed that these inhibitors had a much higher affinity (i.e., a lower Ki*) for plant urease than for microbial urease. Measurement of rate constants for formation (kon) and decay (koff) of E · I* showed that, whereas kon varied greatly with the different inhibitors and ureases, koff was constant for the phosphoroamides tested and had a characteristic value for each urease. The half-life of E · I* (30°C; pH 7 THAM buffer) for the plant urease was much longer than that for the microbial urease, and this difference largely accounted for the much higher values of Ki* (koff/kon) observed with microbial urease.

Effect of nickel deficiency in soybeans on the phytotoxicity of foliar-applied urea

The leaf-tip necrosis commonly observed after foliar fertilization of soybean [Glycine max (L.) Merr.] plants with urea is usually attributed to ammonia formed through hydrolysis of urea by plant urease. We recently found, however, that although addition of a urease inhibitor (phenylphosphorodiamidate) to foliar-applied urea increased the urea content and decreased the ammonia content and urease activity of soybean leaves, it increased the leaf-tip necrosis observed after foliar fertilization. We concluded that this necrosis was due to accumulation of toxic amounts of urea rather than formation of toxic amounts of ammonia. To confirm this conclusion, we measured the urea content, urease activity, and leaf-tep necrosis of leaves of soybean plants treated with urea after growth of the plants in nutrient solutions containing different amounts of nickel (Ni), which is an essential component of urease. We found that the urease activity of these leaves decreased, and that their urea content and leaf-tip necrosis increased, with decrease in the Ni content of the nutrient solution. Besides supporting the conclusion that the leaf-tip necrosis observed after foliar fertilization of soybean with urea is due to accumulation of toxic amounts of urea in the soybean leaves, these observations indicate that Ni-deficient plants may have a lower urease activity than plants that are not deficient in Ni and may therefore be more susceptible to leaf burn when foliar-fertilized with urea.