Impacts of organic and inorganic fertilizers on nitrification in a cold climate soil are linked to the bacterial ammonia oxidizer community

The microbiology underpinning soil nitrogen cycling in northeast China remains poorly understood. These agricultural systems are typified by widely contrasting temperature, ranging from −40 to 38°C. In a long-term site in this region, the impacts of mineral and organic fertilizer amendments on potential nitrification rate (PNR) were determined. PNR was found to be suppressed by long-term mineral fertilizer treatment but enhanced by manure treatment. The abundance and structure of ammonia-oxidizing bacterial (AOB) and archaeal (AOA) communities were assessed using quantitative polymerase chain reaction and denaturing gradient gel electrophoresis techniques. The abundance of AOA was reduced by all fertilizer treatments, while the opposite response was measured for AOB, leading to a six- to 60-fold reduction in AOA/AOB ratio. The community structure of AOA exhibited little variation across fertilization treatments, whereas the structure of the AOB community was highly responsive. PNR was correlated with community structure of AOB rather than that of AOA. Variation in the community structure of AOB was linked to soil pH, total carbon, and nitrogen contents induced by different long-term fertilization regimes. The results suggest that manure amendment establishes conditions which select for an AOB community type which recovers mineral fertilizer-suppressed soil nitrification.     

Recovery of Soil Ammonia Oxidation After Long-Term Zinc Exposure Is Not Related to the Richness of the Bacterial Nitrifying Community

A soil sterilization–reinoculation approach was used to manipulate soil microbial diversity and to assess the effect of the diversity of the ammonia-oxidizing bacteria (AOB) on the recovery of the nitrifying community to metal stress (zinc). Gamma-irradiated soil was inoculated with 13 different combinations of up to 22 different soils collected worldwide to create varying degrees of AOB diversity. Two months after inoculation, AOB amoA DGGE based diversity (weighted richness) varied more than 10-fold among the 13 treatments, the largest value observed where the number of inocula had been largest. Subsequently, the 13 treatments were either or not amended with ZnCl₂. Initially, Zn amendment completely inhibited nitrification. After 6 months of Zn exposure, recovery of the potential nitrification activity in the Zn amended soils ranged from <10 % to >100 % of the potential nitrification activity in the corresponding non-amended soils. This recovery was neither related to DGGE-based indices of AOB diversity nor to the AOB abundance assessed 2 months after inoculation (p?>?0.05). However, recovery was significantly related (r?=?0.75) to the potential nitrification rate before Zn amendment and only weakly to the number of soil inocula used in the treatments (r?=?0.46). The lack of clear effects of AOB diversity on recovery may be related to an inherently sufficient diversity and functional redundancy of AOB communities in soil. Our data indicate that potential microbial activity can be a significant factor in recovery.     

Does long-term fertilization treatment affect the response of soil ammonia-oxidizing bacterial communities to Zn contamination?

Experiments were conducted to examine the effects of long-term fertilization and acute Zn toxicity on the size, nitrification activity and community structure of autotrophic ammonia-oxidizing populations of the β-subgroup of the class Proteobacteria in arable soils. Plots under different long-term fertilization regimes were sampled, and then different concentrations of ZnCl₂ were spiked into soil samples for 8 weeks. It was found that long-term fertilization significantly increased nitrification rates and population size, and there was a positive correlation between them. A shift in the composition of AOB was also detected in samples fertilized with mineral N fertilizer (NPK) and organic matter (OM) as compared to unfertilized sample. EC50 values suggested that there was no significant difference in Zn toxicity to nitrification rates among the three fertilization treatments. Long-term fertilization did not improve the resilience of AOB activity to Zn toxicity.     

Development of Metal Tolerance in Soil Bacterial Communities Exposed to Experimentally Increased Metal Levels

The development of metal tolerance in soil bacterial communities exposed to different heavy metals was examined under laboratory conditions. An agricultural soil amended with different Zn concentrations was studied most intensively, and measurements were made over a 28-month incubation period by means of the thymidine incorporation technique. Tolerance levels were not affected by metal concentrations lower than 2 mmol of Zn kg (dry weight) of soil(sup-1), but above this value, the level of Zn tolerance increased exponentially with the logarithm of the soil Zn concentration. An increased metal tolerance was detected after only 2 days of Zn exposure. Thereafter, stable tolerance values were observed at different sampling times for bacterial communities exposed to up to 8 mmol of Zn kg (dry weight)(sup-1), indicating no changes in tolerance with time. The tolerance of bacterial communities exposed to 32 mmol of Zn kg (dry weight)(sup-1) increased rapidly within the second week of incubation, but then the values remained unchanged until the end of the experiment. Bacterial communities from soil contaminated with 16 mmol of Zn kg (dry weight)(sup-1) showed an increase of the same magnitude, but the increase started later, after 4 months of incubation, and took place for a much longer period (more than 1 year). Cd, Cu, and Ni addition also resulted in metal-tolerant communities, and the level of tolerance increased with prolonged incubations of the soils. The bacterial community at the end of the incubation period also exhibited a lower pH optimum and an increased tolerance to low osmotic potential. The results suggest that the increase in metal tolerance of the community after adding metals can be attributed to an immediate effect due to the death of sensitive species and a later effect due to different competitive abilities and adaptation of surviving bacteria.     

Microbial Biomass, Community Structure and Metal Tolerance of a Naturally Pb-Enriched Forest Soil

The effect of long-term elevated soil Pb levels on soil microbiota was studied at a forest site in Norway, where the soil has been severely contaminated with Pb since the last period of glaciation (several thousand years). Up to 10% Pb (total amount, w/w) has been found in the top layer. The microbial community was drastically affected, as judged from changes in the phospholipid fatty acid (PLFA) pattern. Specific PLFAs that were high in Pb-enriched soil were branched (especially br17:0 and br18:0), whereas PLFAs common in eukaryotic organisms such as fungi (18:2ω6,9 and 20:4) were low compared with levels at adjacent, uncontaminated sites. Congruent changes in the PLFA pattern were found upon analyzing the culturable part of the bacterial community. The high Pb concentrations in the soil resulted in increased tolerance to Pb of the bacterial community, measured using both thymidine incorporation and plate counts. Furthermore, changes in tolerance were correlated to changes in the community structure. The bacterial community of the most contaminated soils showed higher specific activity (thymidine and leucine incorporation rates) and higher culturability than that of control soils. Fungal colony forming units (CFUs) were 10 times lower in the most Pb-enriched soils, the species composition was widely different from that in control soils, and the isolated fungi had high Pb tolerance. The most commonly isolated fungus in Pb-enriched soils was Tolypocladium inflatum. Comparison of isolates from Pb-enriched soil and isolates from unpolluted soils showed that T. inflatum was intrinsically Pb-tolerant, and that the prolonged conditions with high Pb had not selected for any increased tolerance.     

Effects of zinc-smelter emissions on forest soil microflora.

Within 2 km of a zinc (Zn) smelter in Palmerton, Pennsylvania, near the Lehigh Water Gap, up to 13.5% Zn by weight has been measured in the O2 horizon of the soil, and up to 8% Zn in the A1 horizon. The total numbers of bacteria, actinomycetes, and fungi (measured by dilution plate counts) were greatly reduced in the most severely Zn-contaminated soils compared with control soils. The reduction of microbial populations may be a partial cause of the decreased rate of litter decomposition at Lehigh Gap. Growth of most bacteria from control sites was reduced by 100 to 200 muM Zn, most actinomycetes by 100 muM Zn, and most fungi by 100 to 1000 muM Zn in thin-Pablum extract agar (TPab). All the tested actinomycetes and non-spore-forming bacteria isolated from Zn-contaminated Lehigh Gap soils were Zn-tolerant, growing normally in media containing 600-2000 muM Zn. Most fungi, regardless of source, were capable of at least 50% of normal growth at 700 muM Zn. Zinc-tolerant bacteria, actinomycetes, and fungi were readily isolated from low-Zn soils, suggesting that selection for Zn tolerance may proceed rapidly. Acidophilic Mortierella species have been selectively eliminated near the smelter, apparently because of elevated soil pH. Peryronellaea glomerata (Corda) Goidanich and Coniothyrium spp. were found only in the high-Zn soils.     

Root porosity, radial oxygen loss and iron plaque on roots of wetland plants in relation to zinc tolerance and accumulation

Background and aims

Wetland plants have been widely used in constructed wetlands for the clean-up of metal-contaminated waters. This study investigated the relationship between rate of radial oxygen loss (ROL), root porosity, Zn uptake and tolerance, Fe plaque formation in wetland plants.

Methods

A hydroponic experiment and a pot trial with Zn-contaminated soil were conducted to apply different Zn level treatments to various emergent wetland plants.

Results

Significant differences were found between plants in their root porosities, rates of ROL, Zn uptake and Zn tolerance indices in the hydroponic experiment, and concentrations of Fe and Mn on roots and in the rhizosphere in the pot trial. There were significant positive correlations between root porosities, ROL rates, Zn tolerance, Zn, Fe and Mn concentrations on roots and in the rhizosphere. Wetland plants with higher root porosities and ROL tended to have more Fe plaque, higher Zn concentrations on roots and in their rhizospheres, and were more tolerant of Zn toxicity.

Conclusions

Our results suggest that ROL and root porosity play very important roles in Fe plaque formation, Zn uptake and tolerance, and are useful criteria for selecting wetland plants for the phytoremediation of Zn-contaminated waters and soils/sediments.     

Assessment of Pollution-Induced Microbial Community Tolerance to Heavy Metals in Soil Using Ammonia-Oxidizing Bacteria and Biolog Assay

This study attempted to investigate if the tolerance of soil bacterial communities in general, and autotrophic ammonia-oxidizing bacteria (AOB) in particular, evolved as a result of prolonged exposure to metals, and could be used as an indigenous bioindicator for soil metal pollution. A soil contaminated with copper, chromium, and arsenic (CCA) was mixed with an uncontaminated garden soil (GS3) to make five test soils with different metal concentrations. A modified potential ammonium oxidation assay was used to determine the metal tolerance of the AOB community. Tolerance to Cr, Cu, and As was tested at the beginning and after up to 13 months of incubation. Compared with the reference GS3 soil, the five CCA soils showed significantly higher tolerance to Cr no matter which form of Cr (Cr³⁺, CrO₄ ^2?, or Cr₂O₇ ^2?) was tested, and the Cr tolerance correlated with the total soil Cr concentration. However, the tolerance to Cu²⁺, As³⁺, and As⁵⁺ did not differ significantly between the GS3 soil and the five CCA soils. Community level physiological profiles using Biolog microtiter plates were also used to examine the chromate tolerance of the bacterial communities extracted after six months of exposure. Our results showed that the bacterial community tolerance was altered and increased as the soil Cr concentration was increased, indicating that the culturable microbial community and the AOB community responded in a similar manner.     

Effect of Metal-Rich Sludge Amendments on the Soil Microbial Community

The effects of heavy-metal-containing sewage sludge on the soil microbial community were studied in two agricultural soils of different textures, which had been contaminated separately with three predominantly single metals (Cu, Zn, and Ni) at two different levels more than 20 years ago. We compared three community-based microbiological measurements, namely, phospholipid fatty acid (PLFA) analysis to reveal changes in species composition, the Biolog system to indicate metabolic fingerprints of microbial communities, and the thymidine incorporation technique to measure bacterial community tolerance. In the Luddington soil, bacterial community tolerance increased in all metal treatments compared to an unpolluted-sludge-treated control soil. Community tolerance to specific metals increased the most when the same metal was added to the soil; for example, tolerance to Cu increased most in Cu-polluted treatments. A dose-response effect was also evident. There were also indications of cotolerance to metals whose concentration had not been elevated by the sludge treatment. The PLFA pattern changed in all metal treatments, but the interpretation was complicated by the soil moisture content, which also affected the results. The Biolog measurements indicated similar effects of metals and moisture to the PLFA measurements, but due to high variation between replicates, no significant differences compared to the uncontaminated control were found. In the Lee Valley soil, significant increases in community tolerance were found for the high levels of Cu and Zn, while the PLFA pattern was significantly altered for the soils with high levels of Cu, Ni, and Zn. No effects on the Biolog measurements were found in this soil.     

Do ammonia-oxidizing archaea respond to soil Cu contamination similarly asammonia-oxidizing bacteria?

Inhibitory experiments were conducted to investigate the responses of the population sizes of ammonia-oxidizing archaea (AOA) and bacteria (AOB) and the potential nitrification rates (PNRs) to Cu contamination in four Chinese soils. PNR was determined using a substrate-induced nitrification (SIN) assay, and the population size of the nitrifiers represented by amoA gene abundances was quantified using a real-time polymerase chain reaction (qPCR) assay. Both population size and PNR of the ammonia oxidizers reduced considerably at high Cu concentrations in all the soils. Bacterial amoA gene abundance was reduced by from 107-fold (Hailun soil) to more than 232-fold (Hangzhou soil) at the highest Cu concentrations (2,400 mg kg^?1 Cu for Hailun, Langfang and Guangzhou soils and 1,600 mg kg^?1 Cu for Hangzhou soil), while reduction in archaeal amoA gene abundance was from 10-fold (Langfang soil) to 89-fold (Hangzhou soil). AOA seemed more tolerant to Cu contamination than AOB. Nitrification rates were inhibited by more than 50% at a Cu concentration of 600 mg kg^?1, and by more than 90% at the highest Cu concentrations in all soils. These results indicated that both AOA and AOB can be inhibited by toxic metals, highlighting the need to consider the role of AOA in nitrification in soils.     

Simazine application inhibits nitrification and changes the ammonia-oxidizing bacterial communities in a fertilized agricultural soil

s-Triazine herbicides are widely used for weed control, and are persistent in soils. Nitrification is an essential process in the global nitrogen cycle in soil, and involves ammonia-oxidizing Bacteria (AOB) and ammonia-oxidizing Archaea (AOA). In this study, we evaluated the effect of the s-triazine herbicide simazine on the nitrification and on the structure of ammonia-oxidizing microbial communities in a fertilized agricultural soil. The effect of simazine on AOB and AOA were studied by PCR-amplification of amoA genes of nitrifying Bacteria and Archaea in soil microcosms and denaturing gradient gel electrophoresis (DGGE) analyses. Simazine [50?μg g(-1) dry weight soil (d.w.s)] completely inhibited the nitrification processes in the fertilized agricultural soil. The inhibition by simazine of ammonia oxidation observed was similar to the reduction of ammonia oxidation by the nitrification inhibitor acetylene. The application of simazine-affected AOB community DGGE patterns in the agricultural soil amended with ammonium, whereas no significant changes in the AOA community were observed. The DGGE analyses strongly suggest that simazine inhibited Nitrosobacteria and specifically Nitrosospira species. In conclusion, our results suggest that the s-triazine herbicide not only inhibits the target susceptible plants but also inhibits the ammonia oxidation and the AOB in fertilized soils.     

Effects of heavy-metal additions on ammonification and nitrification in soils contaminated with cadmium,lead and zinc

Summary The sensitivity of the mineralization of nitrogen by a range of soils contaminated with heavy metals (up to 340 μg Cd g⁻¹, 7500 μg Pb g⁻¹ and 34000 μg Zn g⁻¹) to the addition of heavy metals in solution were studied using pot incubations (ammonification) and a soil perfusion technique (nitrification). The ammonification of peptone showed little correlation between treatments with Cd, Zn (1000 and 5000 μg g⁻¹) and Pb (10000 and 20000 μg g⁻¹) and origin of the soil. Nitrification was considerably more sensitive to heavy metals than ammonification. All the soils had active, often large, populations of ammonifying and nitrifying organisms which showed substantial similarities between the soils. The rate of nitrifying activity (NO₃−N production) was logrithmic in most cases. The presence of tolerant populations of nitrifying organisms in the contaminated soils was demonstrated. Tolerance was also eventually acquired after a longer lag phase, by the non-contaminated soil populations although the rate of activity was often reduced. Metals added in solution were adsorbed by the soil within 4 hours. Differences in toxicity between metal salts (chlorides, sulphates and acetate) were attributed to the amount left in solution. However, in many instances, acetate was found to stimulate all the stages in the mineralisation of nitrogen.     

变性梯度凝胶电泳法研究我国不同土壤氨氧化细菌群落组成及活性

实验选用了我国3种不同土壤研究土壤硝化活性、硝化细菌数量,并使用变性梯度凝胶电泳(DGGE)的方法研究了不同土壤中氨氧化细菌(AOB)区系变化。通过28d的土壤培养实验研究发现,潮土具有最强的硝化势,几乎100%的铵态氮转化为硝态氮;而红壤中的硝化势最弱,只有4.9%的铵态氮转化为硝态氮。对这3种土壤硝化细菌进行计数发现,3种土壤氨氧化菌数量差异显著,而3种土壤亚硝酸氧化菌(NOB)处于一个数量级。采用氨氧化菌功能基因amoA(氨单加氧酶ammoniamonooxygenase)特异PCR结合DGGE的方法对土壤氨氧化菌区系进行分析。红壤有4个氨氧化菌种属,与潮土和黄泥土没有共同的氨氧化菌种属。4个种属中两个是与潮土和黄泥土亲源性比较远的,特有的氨氧化菌种属,这两个种属与已知的Nitrosospira属的cluster3bz97838和Nitrosospira属的cluster3aAF353263亲源性比较近。潮土有5个氨氧化菌种属,潮土与黄泥土有两个共同的氨氧化菌种属,这两个种属中的一个是潮土和黄泥土特有的,与其他氨氧化菌种属亲源性比较远的氨氧化菌种属,这个种属与已知的Nitrosospira属的cluster3bZ97849亲源性比较近。黄泥土有4个氨氧化菌种属,除了与潮土共有的一个种属是两种土壤特有的氨氧化菌种属外,黄泥土还有一个与其他氨氧化菌种属亲源性比较远的,黄泥土特有的种属,与Nitrosospira属的cluster3aAF353263亲源性很近。3种土壤中分离到的硝化细菌表现出不同的硝化能力。实验结果表明,以amoA基因为目标的PCR-DGGE是比以16SrDNA为目标的PCR-DGGE更有效的研究氨氧化菌种群的方法;3种土壤的氨氧化菌种群差异显著,尤其是红壤的氨氧化菌种群与另外两种土壤差异明显,这种差异可能与红壤的低pH条件对氨氧化菌种群的长期选择有关;3种土壤中的硝化活性与土壤中的硝化细菌数量没有显著相关,可能由于3种土壤差异显著的土壤环境对硝化活性的影响造成。因此在对不同土壤硝化细菌进行研究时不仅需要对硝化细菌数量进行研究,还需要研究不同土壤中硝化细菌的种属及不同土壤环境条件对硝化细菌硝化活性的影响。     

Root development of the Zinc-hyperaccumulator plant Thlaspi caerulescens as affected by metal origin,content and localization in soil

Penetration into and exploitation of contaminated soils by roots of hyperaccumulator plants is a prerequisite for efficient removal of heavy metals, i.e. efficacy of phytoextraction. This work was undertaken to study the development of roots of the Zn-hyperaccumulator Thlaspi caerulescens under various conditions of soil contamination. Rhizoboxes were constructed with a removable plastic front cover, and filled with soils containing different amounts and forms of metals (Zn, Cd and Pb). Treatments were: homogeneous soil profile, superposition of three layers, inclusion of contaminated soil into uncontaminated soil, or inclusion of uncontaminated soil into uniformly contaminated soil. Four seedlings were transplanted into each rhizobox, and development of the root system was periodically recorded for 133 days. At harvest, the biomass and size of the rosette of aerial parts were determined. The aerial biomass/root length fraction as well as the kinetics of root development varied according to the presence and localization of Zn. The distribution and morphology of roots at harvest were strongly dependent upon the metal content and form in soil. Roots exhibited a high affinity for the Zn-contaminated patches and showed two distinct morphologies according to the concentration of Zn in soil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Metagenomic and network analyses reveal key players in nitrification in upland agricultural soils

Nitrification, a key step in soil nitrogen cycling, is a biologically mediated process crucial to the ecological environment. However, how nitrifiers drive nitrification under different soil properties and climatic factors at large spatial scales is poorly understood. Here, using metagenomic sequencing and network-based approaches, we identified key nitrifying species of upland agricultural soils in northern China, which spans a wide range of climates and geographic distances. We found that potential nitrification rates (PNRs) varied in different soils and were positively correlated with soil pH (5.42–8.46) and mean annual temperature (MAT) and negatively correlated with the C/N ratio. Network analysis revealed that one module (module 3) was significantly correlated with PNR. In this module, 16 dominant nodes were associated with AOB Nitrosomonas and most nodes were significantly correlated with environmental factors, suggesting that abiotic conditions are important for determining the assembly of these key nitrifiers. Our study advanced the understanding of the key nitrifying populations and their environmental drivers in upland agricultural soil across different soil and climate types.     

High abundance of ammonia-oxidizing archaea in acidified subtropical forest soils in southern China after long-term N deposition

Nitrification has been believed to be performed only by autotrophic ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) until the recent discovery of ammonia-oxidizing archaea (AOA). Meanwhile, it has been questioned whether AOB are significantly responsible for NH(3) oxidation in acidic forest soils. Here, we investigated nitrifying communities and their activity in highly acidified soils of three subtropical forests in southern China that had received chronic high atmospheric N deposition. Nitrifying communities were analyzed using PCR- and culture (most probable number)-based approaches. Nitrification activity was analyzed by measuring gross soil nitrification rates using a (15) N isotope dilution technique. AOB were not detected in the three forest soils: neither via PCR of 16S rRNA and ammonia monooxygenase (amoA) genes nor via culture-based approaches. In contrast, an extraordinary abundance of the putative archaeal amoA was detected (3.2?×?10(8) -1.2?×?10(9) g?soil(-1) ). Moreover, this abundance was correlated with gross soil nitrification rates. This indicates that amoA-possessing archaea rather than bacteria were predominantly responsible for nitrification of the soils. Furthermore, sequences of the genus Nitrospira, a dominant group of soil NOB, were detected. Thus, nitrification of acidified subtropical forest soils in southern China could be performed by a combination of AOA and NOB.     

Biofortification of zinc in wheat grain by the application of sewage sludge

Background and aims

Increasing the concentrations of the essential micronutrient Zn in staple crops like grain is desirable for human nutrition. We investigated the long-term ability of municipal treatment works sewage sludge, liquid sewage sludge and ZnCO₃ applied to soils to increase Zn in in wheat grain (Triticum aestivum L.) in a number of field experiments conducted on different soils.

Methods

We used six long-term field experiments that were set up on contrasting soils in England and the target applications were built up between 1994 and 1997. Topsoil samples and harvested grain samples were taken and air dried in 1999, 2001, 2003 and 2005. Relationships between grain Zn concentrations and soil properties and changes with time were examined.

Results

Wheat grain Zn concentrations increased with soil Zn concentrations in a similar log-log relationship with all of the Zn sources tested. Comparing total or extractable Zn in soil as explanatory factors showed little benefit of using extractable Zn measurements to predict grain concentrations over total Zn. Additional factors such as soil pH or organic carbon did not explain much more of the variation in grain Zn in our experiments. However, grain Zn concentrations did not respond at all at a site with pH 7.7.

Conclusions

Sewage sludge applications to soil can increase grain Zn concentrations for at least 2 to 8?years after application and has similar effectiveness to ZnCO₃.     

Initial Copper Stress Strengthens the Resistance of Soil Microorganisms to a Subsequent Copper Stress

To improve the prediction of essential ecosystem functioning under future environmental disturbances, it is of significance to identify responses of soil microorganisms to environmental stresses. In this study, we collected polluted soil samples from field plots with eight copper levels ranging from 0 to 3,200 mg Cu kg^?1 soil. Then, the soils with 0 and 3,200 mg Cu kg^?1 were selected to construct a microcosm experiment. Four treatments were set up including Cu0-C and Cu3200-C without further Cu addition, and Cu0-A and Cu3200-A with addition of 57.5 mg Cu kg^?1 soil. We measured substrate-induced respiration (SIR) and potential nitrification rate (PNR). Furthermore, the abundance of bacterial, archaeal 16S rRNA genes, ammonia-oxidizing bacteria and archaea amoA genes were determined through quantitative PCR. The soil microbial communities were investigated by terminal restriction fragment length polymorphism (T-RFLP). For the field samples, the SIR and PNR as well as the abundance of soil microorganisms varied significantly between eight copper levels. Soil microbial communities highly differed between the low and high copper stress. In the microcosm experiment, the PNR and SIR both recovered while the abundance of soil microorganisms varied irregularly during the 90-day incubation. The differences of microbial communities measured by pairwise Bray–Curtis dissimilarities between Cu0-A and Cu0-C on day 0 were significantly higher after subsequent stress than before. However, the differences of microbial communities between Cu3200-A and Cu3200-C on day 0 changed little between after subsequent stress and before. Therefore, initial copper stress could increase the resistance of soil microorganisms to subsequent copper stress.     

Reaction norms of life history traits in response to zinc in Thlaspi caerulescens from metalliferous and nonmetalliferous sites

* We examined phenotypic plasticity of fitness components in response to zinc (Zn) in the Zn hyperaccumulator, Thlaspi caerulescens. * Two populations from Zn-enriched soils (M) and two populations from normal soils (NM) were grown in pots at three Zn concentrations (0, 1000 and 8000 mg kg(-1) Zn), for an entire life cycle. Growth, Zn accumulation and fitness components were assessed. * Based on vegetative growth, M and NM populations had similar Zn tolerance at 1000 mg kg(-1) Zn. However, reproductive output was markedly decreased in NM at 1000 and 8000 mg kg(-1) Zn. In M populations, Zn did not affect fitness. However, low Zn status enhanced reproductive output in year 1 compared with year 2 and decreased survival after the first flowering season. * M populations are able to achieve equal fitness across a broad range of Zn concentrations in soil by different combinations of fecundity and longevity. No cost of higher tolerance was demonstrated in M populations. Reproductive traits appeared to be a more sensitive indicator of tolerance than vegetative growth.     

Phospholipid Fatty Acid Composition and Heavy Metal Tolerance of Soil Microbial Communities along Two Heavy Metal-Polluted Gradients in Coniferous Forests

The effects of long-term heavy metal deposition on microbial community structure and the level of bacterial community tolerance were studied along two different gradients in Scandinavian coniferous forest soils. One was near the Harjavalta smelter in Finland, and one was at Ronnskar in Sweden. Phospholipid fatty acid (PLFA) analysis revealed a gradual change in soil microbial communities along both pollution gradients, and most of the individual PLFAs changed similarly to metal pollution at both sites. The relative quantities of the PLFAs br18:0, br17:0, i16:0, and i16:1 increased with increasing heavy metal concentration, while those of 20:4 and 18:2(omega)6, which is a predominant PLFA in many fungi, decreased. The fungal part of the microbial biomass was found to be more sensitive to heavy metals. This resulted in a decreased fungal/bacterial biomass ratio along the pollution gradient towards the smelters. The thymidine incorporation technique was used to study the heavy metal tolerance of the bacteria. The bacterial community at the Harjavalta smelter, exposed mainly to Cu deposition, exhibited an increased tolerance to Cu but not to Cd, Ni, and Zn. At the Ronnskar smelter the deposition consisting of a mixture of metals increased the bacterial community tolerance to all tested metals. Both the PLFA pattern and the bacterial community tolerance were affected at lower soil metal concentrations than were bacterial counts and bacterial activities. At Harjavalta the increased Cu tolerance of the bacteria and the change in the PLFA pattern of the microbial community were found at the same soil Cu concentrations. This indicated that the altered PLFA pattern was at least partly due to an altered, more metal-tolerant bacterial community. At Ronnskar, where the PLFA data varied more, a correlation between bacterial community tolerance and an altered PLFA pattern was found up to 10 to 15 km from the smelter. Farther away changes in the PLFA pattern could not be explained by an increased community tolerance to metals.