Intraspecific differences in nitrogen assimilating enzymes in spinach under contrasting forms of nitrogen supply 1

Intraspecific differences in the activities of nitrate reductase (NR), glutamine synthetase (GS), NADH dependent glutamate synthase (NADH‐GOGAT), and glutamate dehydrogenase (GDH) under contrasting forms of nitrogen (N) supply were studied in tissues of three spinach (Spinacia oleracea L.) cultivars. The varieties (Viroflay, Butterflay, and Giant) were smooth, curly and semicurly leaved, respectively. The plants were grown in nutrient solutions containing NO₃ as the sole source of N (100:0) and NO₃ plus NH₄ (80:20). Giant, the NH₄ tolerance of which had been evaluated in growth and on the basis of nutrient uptake, had much higher GS and GDH activities in the roots and higher NR and NADH‐GOGAT levels in the leaves of plants grown on NO₃ and NH₄ than that grown on NO₃ alone. On the level of N assimilating enzymes of Butterflay, mixed N nutrition caused an increase of GDH and NADH‐GOGAT in leaves and roots and at the same time a decrease of GS in the roots and NR in the leaves. An inverse relationship between GS and GDH activities was detected in the leaves and foots of Virofiay grown with both N sources. Finally, Viroflay gave the highest levels of GDH irrespective of the NO₃:NH₄ assayed, whereas the leaves of Giant were GDH deficient in comparison with the other cultivars. In addition, the GS activity approached zero in the roots of spinach cultivars characterized by hardly any NH₄ tolerance, whereas in those of Giant it increased remarkably with the supply of NO₃ plus NH₄.     

Regulation of extracellular protease activity in soil in response to different sources and concentrations of nitrogen and carbon

A large proportion of the nitrogen (N) in soil is in the form of proteinaceous material. Its breakdown requires the activity of extracellular proteases and other decomposing enzymes. The goal of our study was to better understand how carbon (C) and N availability affect soil protease activity. Several aerobic incubations were carried out with ammonium (NH₄⁺) and proteins as N sources and cellulose as the main C source. A strong increase in protease activity was observed when proteins were added, the increase depending on the amount of protein added and its solubility. Protease synthesis was clearly substrate induced, as NH₄⁺ had no effect. During this substrate induced phase, the addition of glucose but not NH₄⁺ resulted in protease repression, indicating that the level of protease synthesis was determined by the need for C rather than N. After 1 month of incubation, protease activity remained relatively constant over time and was closely related to microbial biomass N. Different concentrations of mineral N in soil solution had no direct effect on protease activity. However, during this stationary phase, protease activity could be repressed by glucose and NH₄⁺ in a treatment with low mineral N content while in treatments with a higher N availability no repression was observed. We hypothesize that the need for N determined protease activity in the treatment with limited N availability. The addition of NH₄⁺ allowed for reallocation of C and N away from protease synthesis, leading to the observed decrease in protease activity. The repression by glucose may be attributed to shifts in the pathway of microbial NH₄⁺ assimilation. The results emphasize the close links between the microbially mediated cycles of organic C and N.     

Effects of methyl bromide fumigation,chloropicrin fumigation and steam sterilization on soil nitrogen dynamics and microbial properties in a pot culture experiment

Abstract

The effects of steam sterilization (SS), methyl bromide (MeBr) fumigation and chloropicrin (CP) fumigation on soil N dynamics and microbial properties were evaluated in a pot experiment. All disinfection treatments increased the NH⁺ ₄-N level and inhibited nitrification. The additional NH⁺ ₄-N in the CP treatment probably originated from the decomposition of microbial debris by surviving microbes, while that in the SS treatment was attributable to deamination processes of soil organic N occurring in a less labile fraction in addition to the decomposition of microbial debris. The MeBr fumigation increased the level of NH⁺ ₄-N without changing the soil microbial biomass. Based on the determinations of soil microbial biomass, substrate utilization activity (Biolog method) and microbial community structure (phospholipid fatty acid method), the effects of the MeBr, CP and SS treatments on the microbial community were compared. The MeBr fumigation had relatively mild and short-term effects on microbial biomass and activity, but altered the community structure drastically by promoting the growth of gram-positive bacteria. The CP fumigation had large and long-term impacts on microbial biomass and activity; the community structure remained unaffected except for the gram-negative bacteria. Steam sterilization had severe and persistent effects on all parameters. The severity of the effects decreased in the order SS ≥ CP > MeBr.     

Nitrogen-Metabolizing Enzymes: Effect of Nitrogen Sources and Saline Irrigation

Abstract

Indian mustard (Brassica juncea L. cv. RH-30) was grown under different types and levels of nitrogen (N) sources, i.e. nitrate, ammonical, and nitrate plus ammonical, at 40, 80, and 120 kg ha^{? 1} under green house conditions. The plants were salinized with 8 and 12 dSm^{? 1} at 35 and 55 days after sowing. A progressive inhibition of the activity of enzymes of N metabolism, i.e., nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH), was observed with increasing level of salinity. However, the magnitude of such reductions was lowest at the highest level of N (120 kg ha^{? 1}) as compared with the lowest level (40 kg ha^{? 1}) irrespective of N source. The activity of nitrate-assimilating enzymes (NR and NiR) was maximum with nitrate fertilizer, and minimum with the ammonical form. The alleviation of detrimental effects of salinity on NR and NiR was better with the highest level of N (120 kg ha^{? 1}) in nitrate form as compared with the lowest level of N (40 kg ha^{? 1}). In contrast, the maximum activity of ammonium-assimilating enzymes (GS, GOGAT, and GDH) was observed with the highest level of N (120 kg ha^{? 1}) and the minimum with the nitrate form of N under salinity. These results indicate that despite the high salinity, an increase in the concentration and uptake of N stimulates the activities of nitrate-assimilating enzymes (NR and NiR) as well as of the ammonia-assimilating enzymes (GS, GOGAT, and GDH).     

外源蔗糖对高NH4+胁迫下拟南芥碳氮代谢的影响

以拟南芥野生型、amt1.1和amt1.3为实验材料，采取土培的方法，以正常培养液（4mmol/L NH4+）培养，在20mmol/L NH4+的胁迫下，通过在培养液中添加0%（T1）蔗糖、5%（T2）蔗糖，测定地上部分的鲜重，叶绿素，游离NH4+，可溶性糖，可溶性蛋白，谷氨酰胺合成酶（GS）、谷氨酸脱氢酶（GDH），矿质元素含量等指标，研究外源蔗糖对NH4+胁迫拟南芥碳氮代谢的影响。结果表明，T1处理下，拟南芥生长受到严重的抑制，鲜重，GS，GDH酶活性降低，游离NH4+含量，叶绿素含量，可溶性糖和可溶性蛋白含量增加，植株的N、P、K、Ca的含量增加，Mg、Fe的含量减少。其中col-0在T1处理下受到的抑制比amt1.1和amt1.3更为显著。与T1处理相比较，T2处理增加了拟南芥植株的鲜重，显著提高了可溶性糖和可溶性蛋白含量，提高了GS和GDH的活性；降低了叶绿素和游离NH4+的含量，提高了植株体内的N、P、K、Ca，Mg的含量，降低了植株Fe的含量，其中，外源蔗糖对col-0高NH4+毒害的缓解更为显著。     

Ammonium assimilation in rice based on the occurrence of 15N and inhibition of glutamine synthetase activity

Assimilation of ammonium (NH₄) into free amino acids and total reduced nitrogen (N) was monitored in both roots and shoots of two‐week old rice seedlings supplied with 5 mM 99% (¹⁵NH₄)₂SO₄ in aerated hydroponic culture with or without a 2 h preincubation with 1 mM methionine sulfoximine (MSX), an inhibitor of glutamine synthetase (GS) activity. ¹⁵NH₄ was not assimilated into amino acids when the GS/GOGAT (glutamate synthase) cycle was inhibited by MSX. Inhibition of glutamine synthetase (GS) activity in roots with MSX increased both the amount of NH₄ and the abundance of ¹⁵N labeled NH₄. In contrast, the amount of Gln and Glu, and their proportions as ¹⁵N, decreased in roots when GS activity was inhibited. This research confirms the importance of GS/GOGAT in NH₄ assimilation in rice roots.

¹⁵N‐labeled studies indicate that NH₄ ions incorporated by roots of rice are transformed primarily into glutamine (Gln) and glutamic acid (Glu) before being converted to other amino acids through transamination (15). The formation of amino acids such as aspartic acid (Asp) and alanine (Ala) directly from free NH₄ in roots also has been reported (4,15). Translocation of free NH₄ to plant shoots, based on the concentration of free NH₄ in xylem exudate, has been reported in tomato (13), although NH₄ in shoots primarily originates from nitrate reduction in the shoot. Photorespiration also can contribute to the accumulation of NH₄ in leaves (7).

The GS/GOGAT cycle appears to be primarily responsible for the assimilation of exogenously supplied NH₄ and NH₄ derived from nitrate reduction in leaves, as well as NH₄ derived from photorespiration (2,3,6,8). Genetic evidence cited to support this conclusion includes the lethal effect of photorespiratory conditions on plant mutants deficient in chloroplast‐localized GS and GOGAT activities (2,3,9), and the rapid accumulation of free NH₄ in GS‐deficient mutants under photorespiratory conditions (2,3,5).

The present study was initiated to quantify the in vivo amino acid synthesis in rice roots and shoots by analysis of ¹⁵N labeling, and should provide a more complete understanding of this important system for NH₄ utilization.     

Perennial winged bean yield and nitrogen fixation improvement with soil fertility treatments of a typic eutrustox

The perennial legume, Winged Bean (Psophocarpus tetragonolobus (L) DC), has potential as a high protein food crop for the humid, tropical regions of the world. Edible seed pods, oil seed grain, leaves, flowers, and unique high protein tubers provide abundant nutritious components desirable for improved human diets. However, soil characteristics and fertility levels influence plant growth, yields and nitrogen fixation capability of this legume. Objectives of this study were to determine soil‐plant nutrient influences on vigorous regrowth, pod and tuber yields, nitrogenase (C₂H₂ red.) activity levels, and nodule cytosol components of the perennial Siempre cultivar grown on a Typic Eutrustox during three years, 1978–1980.

Available soil phosphorus was a first limiting plant nutrient during all three years of plant age. Effects of combined 100 mg P with 200 mg K/kg soil were highly significant for every parameter and plant age. Pod and seed yields more than doubled with PK addition compared to the check. Tuber growth, nodule mass and nitrogenase activity levels more than trebled with PK treatments as compared to the check. Both elemental P and K were significantly increased within the nodule cytosol of fertilized plants. Cytosol Na was significantly decreased with soil K additions. The best fit multiple regression was: nitrogenase = 1.99 nodule wt. + 6.34 tuber wt. + 0.39 tuber % N + 5.08 cytosol P + 1.55 cytosol K ‐ 0.45 cytosol Na, R² = 95.5, C.V. = 11.2%. The dominant nodule cytosol enzymes, aspartate aminotransferase (AST) and glutamine synthetase (GS), significantly increased with soil K additions regardless of P treatments. Glutamate dehydrogenase (GDH) and glutamate synthase (GOGAT) also contributed significantly with multiple regression for nitrogenase = 1.07 GS + 2.1 AST + 1.74 GOGAT ‐ 1.76 GDH + 12.89 Ureide, R² = .89, C.V. = 17.3%. Highly significant increases in nodule cytosol ureide composition with K soil additions has interest because of the role as a nitrogenous nonprotein component for many legumes. Increases in growth, nodulation and nitrogenase activity levels resulted with increased K levels of 0, 100, 200 and 300 mg K/kg soil when soil P and Ca were not limiting.     

利用gs1.1和gs1.2突变体研究外源蔗糖对高铵胁迫拟南芥碳氮代谢的影响

以拟南芥野生型Col-0、谷氨酰胺合成酶敲除突变体gs1.1和gs1.2为实验材料,采取土培试验,比较正常培养液(4 mmol/L NH4⁺)培养(CK)、正常培养液(4 mmol/L NH4⁺)下外源添加5%蔗糖(T1)、高NH4⁺胁迫(20 mmol/L)(T2)以及高NH4⁺胁迫(20 mmol/L)下外源添加5%蔗糖(T3)对拟南芥各株系各生理指标的影响;通过测定地上部分的鲜重、叶绿素、游离NH4⁺、可溶性糖、可溶性蛋白、谷氨酰胺合成酶(GS)、谷氨酸脱氢酶(GDH)、矿质元素含量等指标,研究外源蔗糖对NH4⁺胁迫拟南芥碳氮代谢的影响。结果表明,高NH4⁺胁迫下,拟南芥生长受到严重的抑制,鲜重、GS、GDH酶活性降低,游离NH4⁺含量、叶绿素含量、可溶性糖和可溶性蛋白含量增加,植株的N、P、K、Ca的含量增加,Mg、Fe的含量减少,其中gs1.1和gs1.2在高NH4⁺处理下受到的抑制比Col-0更为显著。外源添加5%蔗糖显著缓解了高NH4⁺毒害,提高了可溶性糖和可溶性蛋白含量,提高了GS和GDH的活性,降低了叶绿素和游离NH4⁺的含量,提高了植株体内的N、P、K、Ca,Mg的含量,降低了植株Fe的含量,其中,外源蔗糖对gs1.1和gs1.2高NH4⁺毒害的缓解更为显著。     

Phytotoxic Effects of Residual Glufosinate on the Nitrogen Assimilation of Transplanted Rapeseed Seedlings

Herbicide application is an efficient method to control weed growth in modern agriculture production, but there is concern about the ecological impact of unwanted herbicide residues in the soil. Rapeseed varieties ZS11 and D148 were used to evaluate the phytotoxic effects of residual glufosinate on the assimilation of nitrogen (N) in rapeseed seedlings transplanted to untreated [0 g hm^?2 glufosinate] or treated [450 g hm^?2 and 900 g hm^?2 glufosinate] soils. Glutamine synthetase (GS) and glutamate dehydrogenase (GDH) activities, the contents of ammonium (NH₄⁺), free amino acids (FAA), and soluble protein (SP), and seedling dry weight (DW) were determined at 5, 8, 11, 20, 40, and 70 d post-transplant. Both concentrations of glufosinate induced physiological phytotoxicity on the N assimilation of transplanted seedlings of both varieties, as their leaves and roots presented reduced GS activities and SP contents, and increased GDH activities, and NH₄⁺ and FAA content. Glufosinate phytotoxicity on the N assimilation of transplanted seedlings reached a plateau at 11 to 20 d. further, GDH in roots and GS in leaves were still significantly different at 70 d. Meanwhile, ZS11 might be more sensitive to glufosinate than D148 since ZS11 had more variation than D148 at the same treatment, and the overdose of glufosinate more strongly inhibited N assimilation than the recommended dose. Therefore, it is essential to apply a suitable glufosinate dose to the transplanted variety, to minimize adverse effects on crops and the environment.

Abbreviations: N, Nitrogen; GS, Glutamine synthetase; GDH, glutamate dehydrogenase; NH₄⁺, ammonium; FAA, free amino acids; SP, soluble proteins; DW, Dry weight; ANOVA, one-way analysis of variance; NO₃^?, nitrate; OECD, Organisation for Economic Co-operation and Development; PPT, phosphinothricin; USEPA, United States Environmental Protection Agency     

Relationship between carbon and nitrogen availability and extracellular enzyme activities in soil

Microorganisms rely on extracellular enzymes to break down insoluble organic polymers such as cellulose, protein, and chitin into smaller units for uptake. Our objective was to investigate the factors affecting the relationship between soil extracellular enzyme activities and C and N turnover. Several aerobic incubations were carried out with ammonium (NH₄⁺) and proteins as N sources and cellulose as the main C source. Cellulase (exocellulase and β-glucosidase) activity was positively correlated with the amount of cellulose added, as well as with the availability of N. A decrease in the C to N ratio of the amendments from 40 to 10 resulted in an increase in exocellulase and β-glucosidase activity of 18% and 10%, respectively. Similarly, the activity of protease initially depended on the amount and kind of protein added; later, however, an increase in carbon availability resulted in an elevated protease activity. Initially, protease and cellulase activity were induced by their corresponding substrates and an increase in activity of both enzymes resulted in a proportional increase in carbon dioxide (CO₂) evolution. Over time, however, the level of enzyme activity became increasingly determined by factors other than substrate availability. In addition, N turnover, while initially determined by the amount and kind of N source added, became increasingly dominated by the C to N ratio of the substrates added. Our study showed that even though enzyme activities alone may not be sufficient to describe the decomposition process, they can yield valuable information about the availability of specific organic compounds and their degradation over time.     

Ammonium immobilisation during chloroform fumigation

The assumption in using the chloroform fumigation technique for microbial biomass determination is that microbes are killed or at least inactivated by the treatment. Problems associated with transformations of the N released on or during fumigation have so far only been associated with the fumigation-incubation method. A laboratory and a field study were carried out to investigate the possible N transformations during biomass determination by the fumigation-extraction method. Labelled NH₄NO₃ (either the NO₃⁻, NH₄⁺ or both pools were ¹⁵N enriched) was applied to the soil and biomass determinations made at intervals subsequently. The size and enrichment of the ammonium (NH₄⁺), and nitrate (NO₃⁻) pools were determined before and after chloroform fumigation. The ¹⁵N enrichment of the NH₄⁺ pool after fumigation could only be explained if immobilisation of ammonium occurred at some time during the 24 h fumigation period. The extent of this immobilisation was calculated. In addition, there was evidence that nitrification occurred during the fumigation procedure at the start of the laboratory study and throughout the field study. The laboratory and field study differed mainly in the dynamics related to NO₃⁻ uptake and release. There was evidence for uptake of NO₃⁻ by the microbial biomass with and without utilization. We conclude that the ¹⁵N enrichment in the microbial biomass cannot be accurately determined when N transformations and release of non-utilized N occurs during fumigation. The possible immobilisation of mineral N during fumigation will affect the magnitude of the factor used to convert measured microbial biomass N to actual microbial biomass N in soil.     

Effect of biochar and nitrogen fertilizers on maize seedling growth and enzyme activity relating to nitrification

Maximizing nitrogen use efficiency (NUE) involves synchronizing the interplay between nitrogen preferential crops and the nitrogen transformation pathways of soil. Biochar may benefit specific N-preference crops in relatively unsuitable soil environments; however, experimental data are lacking. This study tested eight treatments, consisting of four nitrogen treatments (N0 = control; N1 = NH₄Cl; N2 = NaNO₃; and N3 = 1:1 ratio of NH₄⁺ and NO₃⁻) each with biochar applied at 0% or 2% (w/w). The results show that biochar and/or nitrogen application enhanced maize seedling biomass and NO₃⁻-based fertilizer resulted in higher seedling biomass than NH₄⁺-based fertilizer. With the application of biochar and NH₄⁺-based fertilizer, maize seedling biomass increased and soil NH₄⁺-N content was significantly reduced compared with NH₄Cl sole application. Correlation analysis and redundancy analysis revealed that SOC content and inorganic nitrogen content were the main factors influencing maize growth and N absorption. Biochar with or without nitrogen fertilizer (except N1 treatment) significantly increased β-1,4-glucosidase (BG) activity. Co-application treatments also resulted in higher vector length, an indicator of C limitation—the increment might add to the risk of microbial C limitation. The activity of ammonia monooxygenase (AMO), a key enzyme in nitrification, decreased with the co-application of biochar and nitrogen, suggesting the alteration of nitrogen transformation.     

Studies on soil fumigation—I: Effects on ammonium,nitrate and phosphate in soil and on the growth,nutrition and yield of wheat

Fumigation of field soil with chloropicrin alone or followed by methyl bromide, each at 220Kg·ha^?1, released 20–30 parts/10⁶ NH⁺₄-N which persisted for 75 days; such fumigation also doubled the amount of bicarbonate-extractable phosphate 28 days after fumigation. Soil fumigation increased both the vegetative and grain yields as well as increasing the content of N in the grain and the content of K and Cl in the tops at ear emergence. Root growth and the phosphate uptake activity of the roots were increased by soil fumigation.     

Significance of organic nitrogen uptake from plant residues by soil microorganisms as affected by carbon and nitrogen availability

Soil microorganisms can use a wide range of nitrogen (N) compounds. When organic N sources are degraded, microorganisms can either take up simple organic molecules directly (direct route), or organic N may be mineralized first and taken up in the form of mineral N (mineralization-immobilization-turnover [MIT] route). To determine the importance of the direct route, a microcosm experiment was carried out. Two types of wheat residue were added to soil samples, including younger residue with a carbon (C) to N ratio of 12 and older residue with a C to N ratio of 29. Between days 1 and 4, the gross N mineralization rate reached 8.4 and 4.0 mg N kg⁻¹ dry soil day⁻¹ in the treatment with younger and older residue, respectively. During the same period, there was no difference in protease activity between the two residue amended treatments. The fact that protease activity was not related to gross N mineralization, even though the products of protease activity are the substrates for N mineralization, suggests that not all organic molecules released from residue or soil N passed through the soil mineral N pool. In fact, when leucine and glycine were added, only 10 and 53% of the amino acid-N, respectively, was mineralized. The fraction of N taken up via the direct route was estimated to be 55 and 62% for the young and older residue, respectively. After 28 days of incubation, the proportion of amino acid-N mineralized had increased especially in the soil amended with older residue, suggesting that the MIT route became increasingly important. This result is supported by an increase in the activities of enzymes responsible for the intracellular assimilation of ammonium (NH₄⁺). Our results suggest that in contrast to what is proposed by many models of soil N cycling, both the direct and MIT routes were operative, with the direct route being the preferred route of residue N uptake. The direct route became less important over time and was more important in soil amended with older residue, suggesting that the direct route is favored by lower mineral N availabilities. An important implication of these findings is that when the direct route is dominant, gross N mineralization underestimates the amount of N made available from the residue.     

Effects of ammonium and nitrate additions on carbon mineralization in wetland soils

Wetlands have been recognized as a soil carbon (C) sink due to low decomposition. As decomposition is largely controlled by the availability of soil nitrogen (N), an elevated anthropogenic N input could influence the C balance in wetlands. However, the effects of the form of N on decomposition are poorly understood. Here, a 54-day laboratory incubation experiment was conducted, with a diel cycle (day: 22 °C for 13 h; night: 17 °C for 11 h) in order to determine how the dominant N form influences the mineralization of soil C in two adjacent wetland soils, with distinct physicochemical characteristics. Three combinations of N compounds were added at three different rates (0, 30, 60 kg N ha⁻¹ yr⁻¹): Ammonium dominant (NH₄Cl + NH₄NO₃); nitrate dominant (NH₄NO₃ + NaNO₃); and ammonium nitrate treatments (NH₄NO₃). In the acidic soil, the CO₂ efflux was reduced with N additions, especially with NH₄NO₃ treatment. In addition, decreases in the microbial enzyme activities (β-glucosidase, N-acetyl-glucosaminidase, phosphatase, and phenol oxidase) and soil pH were observed with NH₄NO₃ and -dominant treatment. Under alkaline conditions, marginal changes in response to N additions were observed in the soil CO₂ efflux, extractable DOC, simple substrate utilization, enzyme activities and pH. A regression analysis revealed that the changes in pH and enzyme activities after fertilization significantly influenced the soil CO₂ efflux. Our findings suggest that the form of N additions could influence the rate of C cycling in wetland soils via biological (enzyme activities) and chemical (pH) changes.     

Effect of Plant Growth Regulators and Different Nitrogen Sources on Glutamate Dehydrogenase Activity

Radish (Raphanus sativus L.) seedlings, treated with various plant growth regulators (PGRs) [viz. kinetin (KiN), gibberellic acid (GA), and abscisic acid(ABA)] were exposed to different nitrogen (N) sources in light and dark condition, and aminative (NADH) and deaminative (NAD⁺) glutamate dehydrogenase (GDH) activities were measured in cotyledons. A differential effect of nitrogen sources, plant growth regulators, and light or dark condition was observed in all the treatments. The NAD‐GDH (deaminating) activity in radish seedlings was only about 10% of aminating activity (irrespective of the PGR treatment). Except with abscisic acid, in all other treatments, either NAD‐GDH or NADH‐GDH activities were more in dark than in light. The amination and deamination reactions also showed different ratios of activity under different N sources (KNO3, NH₄Cl and NH₄NO₃). These data suggest the presence of isoenzymes or conformers of GDH, specific for each tissue, whose activities vary depending on the physiological condition of the tissue. Different energy status of the seedlings during light or dark condition or with PGR treatments may affect the GDH activity differently.     

Gross nitrogen mineralization and nitrification rates and their relationships to enzyme activities and the soil microbial biomass in soils treated with dairy shed effluent and ammonium fertilizer at different water potentials

 Gross N mineralization and nitrification rates and their relationships to microbial biomass C and N and enzyme (protease, deaminase and urease) activities were determined in soils treated with dairy shed effluent (DSE) or NH₄ ⁺ fertilizer (NH₄Cl) at a rate equivalent to 200 kg N ha^–1 at three water potentials (0, –10 and –80 kPa) at 20  °C using a closed incubation technique. After 8, 16, 30, 45, 60 and 90 days of incubation, sub-samples of soil were removed to determine gross N mineralization and nitrification rates, enzyme activities, microbial biomass C and N, and NH₄ ⁺ and NO₃ ^– concentrations. The addition of DSE to the soil resulted in significantly higher gross N mineralization rates (7.0–1.7 μg N g^–1 soil day^–1) than in the control (3.8–1.2 μg N g^–1 soil day^–1), particularly during the first 16 days of incubation. This increase in gross mineralization rate occurred because of the presence of readily mineralizable organic substrates with low C : N ratios, and stimulated soil microbial and enzymatic activities by the organic C and nutrients in the DSE. The addition of NH₄Cl did not increase the gross N mineralization rate, probably because of the lack of readily available organic C and/or a possible adverse effect of the high NH₄ ⁺ concentration on microbial activity. However, nitrification rates were highest in the NH₄Cl-treated soil, followed by DSE-treated soil and then the control. Soil microbial biomass, protease, deaminase and urease activities were significantly increased immediately after the addition of DSE and then declined gradually with time. The increased soil microbial biomass was probably due to the increased available C substrate and nutrients stimulating soil microbial growth, and this in turn resulted in higher enzyme activities. NH₄Cl had a minimal impact on the soil microbial biomass and enzyme activities, possibly because of the lack of readily available C substrates. The optimum soil water potential for gross N mineralization and nitrification rates, microbial and enzyme activities was –10 kPa compared with –80 kPa and 0 kPa. Gross N mineralization rates were positively correlated with soil microbial biomass N and protease and urease activities in the DSE-treated soil, but no such correlations were found in the NH₄Cl-treated soil. The enzyme activities were also positively correlated with each other and with soil microbial biomass C and N. The forms of N and the different water potentials had a significant effect on the correlation coefficients. Stepwise regression analysis showed that protease was the variable that most frequently accounted for the variations of gross N mineralization rate when included in the equation, and has the potential to be used as one of the predictors for N mineralization. Received: 10 March 1998     

Evaluation of biological and chemical nitrogen indices for predicting nitrogen-supplying capacity of paddy soils in the Taihu Lake region,China

Simple and rapid chemical indices of soil nitrogen (N)-supplying capacity are necessary for fertilizer recommendations. In this study, pot experiment involving rice, anaerobic incubation, and chemical analysis were conducted for paddy soils collected from nine locations in the Taihu Lake region of China. The paddy soils showed large variability in N-supplying capacity as indicated by the total N uptake (TNU) by rice plants in a pot experiment, which ranged from 639.7 to 1,046.2 mg N pot⁻¹ at maturity stage, representing 5.8% of the total soil N on average. Anaerobic incubation for 3, 14, 28, and 112 days all resulted in a significant (P < 0.01) correlation between cumulative mineral NH₄⁺-N and TNU, but generally better correlations were obtained with increasing incubation time. Soil organic C, total soil N, microbial C, and ultraviolet absorbance of NaHCO₃ extract at 205 and 260 nm revealed no clear relationship with TNU or cumulative mineral NH₄⁺-N. Soil C/N ratio, acid KMnO₄-NH₄⁺-N, alkaline KMnO₄-NH₄⁺-N, phosphate–borate buffer extractable NH₄⁺-N (PB-NH₄⁺-N), phosphate–borate buffer hydrolyzable NH₄⁺-N (PB_HYDR-NH₄⁺-N) and hot KCl extractable NH₄⁺-N (H_KCl−NH₄⁺-N) were all significantly (P < 0.05) related to TNU and cumulative mineral NH₄⁺-N of long-term incubation (>28 days). However, the best chemical index of soil N-supplying capacity was the soil C/N ratio, which showed the highest correlation with TNU at maturity stage (R = −0.929, P < 0.001) and cumulative mineral NH₄⁺-N (R = −0.971, P < 0.001). Acid KMnO₄-NH₄⁺-N plus native soil NH₄⁺-N produced similar, but slightly worse predictions of soil N-supplying capacity than the soil C/N ratio.     

Soil fertility effects on growth,nitrogen fixation,nodule enzyme activity and xylem exudates of lablab purpureus (L.) sweet,grown on a typic eutrustox

Abstract

The Lablab or Hyacinth bean, Lablab purpureus (L.) Sweet, is a drought tolerant grain‐forage legume widely grown within a wide range of neotropical regions of the world. It has been an important cultivated food grain crop for many centuries within extensive warm humid‐subhumid climatic areas and grown on widely different Ultisol and Oxisol soil types. The objective of these studies was to determine soil fertility effects of Ca, P, and K factorial combinations applied to a Typic Eutrustox on growth, nodulation, and nitrogen fixation of Lablab bean. Stem and nodule xylem components were determined along with soil pH and nitrification within the nodule rhizosphere. Highly significant increases resulted with P fertilization for stem, leaf, seed, nodulation, and nitrogenase activity. With the exception of nitrogenase, all were also increased with K additions. Both stem and seed were significantly increased with Ca treatments. Highly significant increases for total plant N content resulted with P, K, Ca, and P x K interaction. Allantoinates were initially dominant components of stem xylem exudate following decapitation but were not detected after 5 days. Amino‐amide composition remained relatively constant. Nitrates increased from 1.6 to 81.8 ug N ml^‐1 exudate during the 8 day collection period. Soil nitrate levels adjunct to decomposing nodules increased from 4.2 to 1661.1 ug g^‐1 soil. Initial high levels of ureide transforming enzymes ALTN, ALTC, and URC were not detected after 4 days. Amine‐amide enzymes AST, GS, GOGAT, and GDH were relatively constant with nitrate reductase increasing from 0.12 to 9.35 IU ml^‐1 xylem exudate during the 8 d period. Nodule xylem export components were dominated by ureides, 429.8 umol, and amines 30.3 umol ml^‐1. Enzyme activity levels were highest for AST 22.17 and GS 13.25 IU ml^‐1 with the ureide enzymes URC 2.24, ALTN 0.26, and ALTC 0.15 IU ml^‐1 . Nodule exudate composition was K 422.0, P 63.4, Ca 53.8, Mg 42.8, and Na 25.6 ug ml^‐1.     

不同铵硝配比对弱光下白菜氮素吸收及相关酶的影响

以黑色遮阳网覆盖模仿弱光环境, 使光照强度为自然光的20%左右, 以自然光照为对照, 采用精确控制水培溶液氮素营养, 研究NH₄⁺-N/NO₃^--N 比例分别为0/100、25/75、50/50、75/25、100/0 对弱光下白菜氮代谢及硝酸还原酶和谷氨酰胺合成酶活性的影响。结果表明, 弱光下, 白菜的鲜重及叶片总氮量以NH₄⁺-N/NO₃^--N 比为25/75 时最大, NH₄⁺-N/NO₃^--N 比为100/0 时最低。随弱光处理的进行, 白菜叶片中硝酸还原酶活性及谷氨酰胺合成酶活性均呈下降趋势, 但NH₄⁺-N/NO₃^--N 比为25/75 时, 可维持叶片内较高的硝酸还原酶活性及谷氨酰胺合成酶活性。试验表明, NH₄⁺-N/NO₃^--N 比25/75 是白菜在弱光下生长的较适宜氮素形态配比。