Attempts to derive EC50 values for heavy metals from land-applied Cu-, Ni-, and Zn-spiked sewage sludge

We established a field trial to assess the impacts on soil biological properties of application of heavy metal-spiked sewage sludge, with the aim of determining toxicity threshold concentrations of heavy metals in soil. Plots were treated with sludges containing increasing concentrations of Cu, Ni and Zn in order to raise the metal concentrations in the soil by 0-200 mg Cu kg⁻¹, 0-60 mg Ni kg⁻¹ and 0-400 mg Zn kg⁻¹, and were then cultivated and sown in ryegrass-clover pasture and monitored annually for 6 years. All biological properties measured (soil basal respiration, microbial biomass C, and sulphatase enzyme activities), except phosphatase activity, increased in all plots over the duration of the experiment. Consequently, it was only possible to assess effects of heavy metals across time if, each year, all data for each metal were normalised by expressing them as percentages of the activities measured in an un-sludged control plot. When this was done, no significant effects of increasing heavy-metal concentrations on basal respiration, microbial biomass C or respiratory quotient (qCO₂) were observed, although total Cu and soil solution Cu were significantly negatively related to microbial biomass C when it was expressed as a proportion of soil total C. None of the properties measured were affected by increasing Ni concentrations. Phosphatase and sulphatase activities were significantly negatively related to increasing Zn concentrations, but not usually to increasing Cu unless they were expressed as a proportion of total C. A sigmoidal dose-response model was used to calculate EC₂₀ and EC₅₀ values using the normalised data, but generally, the model parameters had very large 95% confidence intervals and/or the fits to the model had small R² values. The factors primarily responsible for confounding these results were site and sample variations not accounted for by the normalisation process and the absence of any data points at metal concentrations beyond the calculated EC₅₀ values. In the few instances where reasonable EC₂₀ values could be calculated, they were relatively consistent across properties, e.g., EC₂₀ for total Zn and phosphatase (330 mg kg⁻¹), total Zn and sulphatase (310 mg kg⁻¹), and EC₂₀ for total Cu and sulphatase (140 mg kg⁻¹) and total Cu and microbial biomass C (140 mg kg⁻¹), when both sulphatase and microbial biomass C were expressed as a proportion of total C. Our results suggest that Cu and Zn at the upper concentrations used in this experiment were possibly having adverse effects on some soil biological properties. However, much higher metal concentrations will be needed to accurately calculate EC₂₀ and EC₅₀ and this may not be easily achievable without many applications of sewage sludge, even if the sludge is spiked with heavy metals.     

Enzyme activities along a climatic transect in the Judean Desert

Soil enzymes have an important influence on nutrient cycling. We examined spatial and temporal patterns in dehydrogenase, arylsulfatase, alkaline and acid phosphatase activities, and their relationships with organic carbon and microbial biomass nitrogen at three sites in Israel representing different climatic regions: Mediterranean (humid), mildly arid and arid. The sites were selected along a climatic transect from the Judean Mountains in the west to the Dead Sea in the east of Israel. With increasing aridity, soil organic carbon, soil microbial biomass nitrogen, dehydrogenase, phosphatase and different pools of arylsulfatase activities decreased significantly. A sharp change in enzyme activities existed between 260- and 120-mm mean annual rainfall. The arylsulfatase activity of the microbial biomass in the 0–2- and 5–10-cm soil layers usually accounted for more than 50% of the total activity, and the fraction of total activity in the 0–2-cm soil layer of the arid sites was significantly greater than that of the humid site. Dehydrogenase and total and microbial biomass arylsulfatase activities were sensitive indicators of the climatic change along the transect. At the humid and mildly arid sites, the activities of dehydrogenase were less in the winter than in the summer and spring, whereas total and microbial biomass arylsulfatase activities were less in both summer and winter. At the arid site, lower values were observed in the summer at 0–2-cm soil depth. At all sites, lower alkaline phosphatase activities at 0–2 cm were observed in the summer, but there were no significant seasonal differences in acid phosphatase activities. These different seasonal patterns of enzyme activities are attributed to the enzyme source, and specific seasonal soil moisture and temperature conditions at the studied sites. The low dehydrogenase and microbial biomass arylsulfatase activities in the winter at the humid and mildly arid sites are explained by the cold and wet soil conditions, and the low enzyme activity in the summer at the arid site is attributed to the dry and hot soil conditions.     

The use of polyethylene mulches to change soil microclimate as revealed by enzyme activity and biomass nitrogen,sulphur and phosphorus

Summary The effects of white, black and clear polyethylene mulches on temporal fluctuations in protease, sulphatase and phosphatase activities and levels of available and biomass N, S and P in raised beds of soil under a strawberry crop were investigated under field conditions. During spring, summer and early autumn, clear and, to a lesser extent black, polyethylene mulch increased both maximum and minimum mean monthly temperatures over those recorded with white mulch or no mulch. During summer and autumn, soil moisture content measured at monthly intervals was higher under mulched than unmulched conditions. Levels of extractable nitrate and sulphate in soils during autumn followed the order: clear mulch = black mulch > white mulch > no mulch. However, there were no significant differences in levels of enzyme activity or concentrations of biomass N, S or P between treatments. There were significant seasonal fluctuations in estimates of biomass N, S and P, but there was no close relationship between fluctuations in any estimate. This discrepancy was attributed to errors in the chloroform fumigation technique. Enzyme activities showed significant temporal fluctuations but the three enzymes did not show similar times of minimum and maximum activity. The complexity of interactions between soil microbial and biochemical properties and nutrient availability was exemplified by the absence of a discernible relationship between seasonal fluctuations in enzyme activities and levels of biomass N, S and P and/or levels of extractable mineral N, S and P in the soil. Strawberry fruit yields and vegetative growth followed the order : clear mulch = black mulch>white mulch>no mulch. Clear, and to a lesser extent black mulch increased the percentage of total yields produced in the first 2 months of cropping.     

棉花抗黄萎病生理与生化机制研究

本文概述了棉花抗黄萎病的特点与规律,指出棉花植株是通过自身组织结构的强化、植物抗毒素的合成以及微生物之间的互作三方面应对黄萎病菌的侵染。认为棉花植株主要通过促进木质素生成和增加积累量以强化组织结构。在棉花抗病反应中以植物抗毒素的合成为主要途径,包括萜类、酶类和激素类3类化学物质,其中酶类物质如几丁质酶已经在很多植物中被证明具有抗病作用,但是其具体作用途径仍不清楚。土壤中某些微生物也可以对黄萎病菌产生作用。重点讨论了在棉花抗黄萎病信号通路中重要分子磷酸激酶和磷酸酯酶在植株抗性防卫中起着级联调控开关的作用,在现有信号调控研究基础上其在抗病机制中可能的参与途径。对目前棉花抗黄萎病分子育种研究中存在的主要问题和可能的解决途径进行了探讨,展望了其发展的方向。     

酒石酸对黏粒矿物上酸性磷酸酶吸附及活性的影响

采用平衡批处理法,研究了模拟根系分泌物——酒石酸溶液的浓度(0~80 mmol L-1)、pH(2.5~7.0)对酸性磷酸酶在针铁矿、高岭石及黄棕壤和砖红壤胶体(<2μm)上的吸附及比活性影响。结果表明,当酒石酸浓度由0 mmol L-1(作为对照)升高至80 mmol L-1时,酸性磷酸酶在供试土壤胶体和矿物表面的吸附量先急剧降低(0~5mmol L-1之间),后逐渐达到平衡;以羟基化表面为主的针铁矿对酸性磷酸酶的吸附百分率受酒石酸浓度的影响最大,吸附百分率从对照的96.2%降至80 mmolL-1酒石酸浓度时的33.7%,其他以层状铝硅酸盐矿物为主的供试土壤胶体和高岭石则受影响较小,吸附百分率从对照的76.3%~60.6%降至80 mmol L-1酒石酸浓度时的56.2%~41.6%。酸性磷酸酶在酒石酸体系中的最大吸附pH点均在该酶的等电点酸侧,这可能与酒石酸对矿物表面的电荷性质改变有关;酒石酸体系中,针铁矿上酸性磷酸酶的比活性远高于其它供试土壤胶体,该酶的最适比活性点随胶体类型的不同而无变化或有所高移。     

Differential interaction of Aspergillus niger and Peniophora lycii phytases with soil particles affects the hydrolysis of inositol phosphates

The effect of the soil environment on the mobility, stability and catalytic activity of phytase from two sources was compared, as these factors have important implications for the efficacy of enzyme function in soil. Phytase from an ascomycete fungus (Aspergillus niger) and a basidiomycete fungus (Peniophora lycii) was added to soil suspensions from three contrasting soils and activities in the solution and solid phase were monitored. The two enzymes were compared because the P. lycii phytase was known to have greater specific activity and a more acidic isoelectric point (pI) than A. niger and therefore predicted to have different adsorption characteristics. When added to soil suspensions buffered at pH 7.5, both phytases remained in solution in all of the soils. In contrast at near natural soil pH (pH 5.5), only the P. lycii phytase remained in solution, while the A. niger phytase was rapidly adsorbed to the soil solid phase. The extent of this adsorption was reduced, however, in a soil-dependent manner by prior addition of bovine serum albumin (BSA) to the soil suspensions. At the natural pH of the soil, the stability of the P. lycii phytase in soil solution was improved under sterile conditions, whereas degradation of the A. niger phytase was unaffected. Subsequently, P. lycii phytase was shown to be more effective at hydrolysing myo-inositol hexakisphosphate added to the soil. Moreover, the P. lycii phytase also hydrolysed more organic phosphate that was endogenous to a range of soils. This research indicates that the physicochemical properties of fungal phytases affect their mobility and temporal stability and their capacity to hydrolyse inositol phosphates in soil environments.     

Performance of para-nitrophenyl phosphate and 4-methylumbelliferyl phosphate as substrate analogues for phosphomonoesterase in soils with different organic matter content

Phosphomonoesterase (PMEase) activity plays a key role in nutrient cycling and is a potential indicator of soil condition and ecosystem stress. We compared para-nitrophenyl phosphate (pNPP) and 4-methylumbelliferyl phosphate (MUP) as substrate analogues for PMEase in 7 natural ecosystem soils and 8 agricultural top soils with contrasting C contents (8.0-414 g kg⁻¹ C) and pH (3.0-7.5). PMEase activities obtained with pNPP (0.05-5 μmol g⁻¹ h⁻¹) were significantly less than activities obtained with MUP (0.9-13 μmol g⁻¹ h⁻¹), especially in soils with a high organic matter content (>130 g kg⁻¹). Only PMEase activities assayed with MUP correlated significantly with total C and total N (r=0.7, P<0.01 all), and pH (r=−0.71, P<0.01). PMEase activities obtained with the two substrate analogues were correlated when expressed on a C-content basis (r=0.8, P<0.001), but not when expressed on an oven-dry soil weight basis. This indicated that interference by organic matter is related to the quantity rather than to the quality of organic matter. Overall, assaying with MUP was more sensitive compared to assaying with pNPP, particularly in the case of high organic and acid soils.     

华北山前平原农田土壤养分循环的酶学调控

土壤酶是推动土壤养分循环的生物催化剂, 研究华北山前平原农田养分循环的酶学调控机制将有助于评价该地区土壤养分状况。试验用土壤取自中国科学院栾城农业生态系统试验站, 分析了土壤脲酶与碱性磷酸酶活性在0~10 cm、10~20 cm、20~30 cm 和30~40 cm 4 个土层中的分布, 研究了土壤粗砂(2 000~200 μm)、粉砂(200~63 μm)、粉粒(63~2 μm)和黏粒(2~0.1 μm) 4 个颗粒组中土壤酶活性与土壤养分之间的相关性。研究结果表明: 土壤脲酶活性与碱性磷酸酶活性均随土壤深度增加而降低, 土壤酶活性随土壤粒径减小而升高。在土壤粗砂和粉砂组分中, 土壤酶活性与养分含量显著相关, 而在土壤粉粒和黏粒组分中, 土壤酶活性与土壤养分含量并不总是显著相关。这些结果说明: 土壤酶活性在土壤养分循环中起到重要的调控作用, 土壤粗砂和粉砂中的土壤酶具有较高的田间实际催化活性, 对土壤养分循环的贡献较大; 与此相反, 土壤粉粒和黏粒中的土壤酶由于土壤无机矿物的吸附作用而失去部分田间催化能力, 降低了其对土壤养分循环的贡献。     

Shrub encroachment does not reduce the activity of some soil enzymes in Mediterranean semiarid grasslands

Shrub encroachment is a worldwide phenomenon with implications for desertification and global change. We evaluated its effects on the activities of urease, phosphatase and β-glucosidase in Mediterranean semiarid grasslands dominated by Stipa tenacissima by sampling 12 sites with and without resprouting shrubs along a climatic gradient. The presence of shrubs affected the evaluated enzymes at different spatial scales. Soils under S. tenacissima tussocks and in bare ground areas devoid of vascular plants had higher values of phosphatase and urease when the shrubs were present. For the β-glucosidase, this effect was site-specific. At the scale of whole plots (30 m × 30 m), shrubs increased soil enzyme activities between 2% (β-glucosidase) and 22% (urease), albeit these differences were significant only in the later case. Our results indicate that shrub encroachment does not reduce the activity of extracellular soil enzymes in S. tenacissima grasslands.     

蔬菜生长期土壤酶活性的变化

利用长期施肥的定位试验对土壤酶活性进行研究,了解当季蔬菜生长期土壤酶活性的变化情 况。种植乌塌菜小区和种植番茄小区土壤酶活性变化趋势基本一致。有机肥的施用可以提高土壤耕层 转化酶、磷酸酶和脲酶活性。