木焦油污染土壤中微生物特性的空间变异性研究

运用地统计学方法研究了木焦油污染土壤中微生物量、微生物群落结构、微生物活性等的空间变异特征.分别采用26种主要磷脂类脂肪酸(PLFA)的总含量(totPLFA)、PLFA的第一主成分和第二主成分(PLFA PC1和PC2)以及土壤培养过程中CO₂-C的累积释放量(Cre)来表征土壤中的微生物量、微生物群落结构以及微生物活性.结果表明,多数微生物特性指标均存在不同程度的空间自相关性,其半变异函数曲线可用带块金效应的球状模型进行拟合.变量的空间相关距离在2.50～16.60 m之间.PLFA PC1、totPLFA和Cre均具有较强的空间依赖性,其相对结构变差（由结构性因素引起的空间变异）分别为82.3%、79.6%和64.7%,而PLFA PC2 不存在明显的空间依赖性.克立格空间插值图表明,样地中存在几处微生物相对密集分布且代谢活性较高的区域,其中优势微生物菌群是由PLFAs 16:1ω7t,cy17:0,18:1ω7 和cy19:0所表征的革兰氏阴性细菌.土壤中主要污染物多环芳烃含量和空间分布是影响微生物特性空间分布格局的重要因素之一.     

Effect of stand age on soil microbial community structure in wolfberry (Lycium barbarum L.) fields

Soil physicochemical properties and microbes are essential in terrestrial ecosystems through their role in cycling mineral compounds and decomposing organic matter. This study examined the effect of stand age on soil physicochemical properties and microbial community structure in wolfberry (Lycium barbarum L.) fields, in order to reveal the mechanism of soil degradation due to long-term stand of L. barbarum. The objective of the study was achieved by phospholipid fatty acid (PLFA) biomarker analysis of soil samples from L. barbarum fields in Zhongning County, Ningxia Province—the origin of L. barbarum. Five stand ages of L. barbarum were selected, < 1, 3, 6, 9, and 12 years (three plots each). The results showed that soil bulk density increased slightly with increasing stand age, while no clear trend was observed in soil pH or total salinity. As the stand age increased, soil organic matter and nutrients first increased before decreasing, with the highest levels being found in year 9. There was an amazing variety of PLFA biomarkers in soil samples at different stand ages. The average concentrations of total, bacterial, fungal, and actinomycete PLFAs in the surface soil initially decreased and then increased, before decreasing with the stand age in summer. The PLFA concentrations of major microbial groups were highest in year 9, with the total PLFA concentrations being 32.97% and 10.67% higher than those in years < 1 and 12, respectively. Higher microbial PLFA concentrations were detected in summer relative to autumn and in the surface relative to the subsurface soil. The highest ratios of Gram-positive to Gram-negative bacterial (G^?/G⁺) and fungal to bacterial (F/B) PLFAs were obtained in year 6, on average 76.09% higher than those at the other four stand ages. The soil environment was most stable in year 6, with no differences between other stand ages. Therefore, soil microbial community structure was strongly influenced by the stand age in year 6 only. The effect of stand age on soil G^?/G⁺ and microbial community structure varied with season and depth; there was little effect for F/B in the 20–40 cm soil layer. Principal component analysis revealed no correlations between microbial PLFA concentrations and total salinity in the soil; negative correlations were noted between soil pH and F/B in summer (P < 0.01), as well as between soil pH and fungal PLFA in autumn (P < 0.05). Moreover, microbial PLFA concentrations were correlated with soil organic matter (mean R = 0.7725), total nitrogen (mean R = 0.8296), total phosphorus (mean R = 0.8175), available nitrogen (mean R = 0.7458), and available phosphorus (mean R = 0.7795) (P < 0.01). On the whole, the soil ecosystem was most stable in year 6, while soil organic matter, nutrients, and microbial PLFA concentrations were maximal in year 9; thereafter, soil fertility indices and microbial concentrations decreased and soil quality declined gradually as the stand age increased. Therefore, farmers should reduce the application rate of fertilizers, especially compound or mixed fertilizers, in L. barbarum fields; organic or bacterial manure can be applied increasingly to improve the soil environment and prolong the economic life of L. barbarum.     

Metabolic quotient of the soil microflora in relation to plant succession

Summary In this study we propose the hypothesis that ecosystem succession is accompanied by a decrease in the metabolic quotient qCO₂ (respiration-to-biomass ratio) of the soil microflora. The qCO₂ is calculated from basal respiration (CO₂-C·h^-1) per unit microbial biomass carbon (C_mier). The hypothesis was tested by studying two primary successions on recessional moraines of the Rotmoos Ferner (Austria) and the Athabasca Glacier (Canada). For both soil seres (0->200 years) it was shown that the qCO₂ decreased with time, which corroborated the hypothesis. In addition, the short term development of the qCO₂ was demonstrated with a revegetation trial. We observed a rise in qCO₂ for the first two years after reclamation, followed by a subsequent decrease.     

Plant community structure, soil properties and microbial characteristics in revegetated quarries

Quarries are an important type of degraded land in southern China requiring ecological improvement and rehabilitation. In this study, plant community structure, soil properties, and microbial biomass and community function were examined at different rehabilitated phases in three quarries, namely Turret Hill Quarry, Lam Tei Quarry and Shek O Quarry, in Hong Kong. Results show that plant species richness and the percentage of native species increased with rehabilitated ages in the three quarries. The highest coverage of woody species was found at older phases, while the lowest woody coverage occurred at younger phases. Soils were strongly to moderately acidic in reaction, and more acidic soils were found in the older than in the younger sites. Organic C as well as total N and P accumulated in soil along with secondary succession in the three quarries, which were positively correlated with woody species richness. Older phases had higher total microbial biomass C and N which were positively correlated with soil organic C, total N and extractable NO₃-N, as well as woody species coverage and native species richness as shown by the biplot of redundancy analysis. Diversity of utilized carbons suggested that metabolic abilities developed gradually with rehabilitation ages in Shek O Quarry, but Turret Hill Quarry and Lam Tei Quarry had similar patterns of carbon source utilization. Principal component analysis further revealed consistent differences in metabolic diversity. Woody coverage and native species richness were significantly correlated with carbon source utilization and functional diversity.     

Mycorrhiza-plant colonization patterns on a subalpine glacier forefront as a model system of primary succession

Lyman glacier in the North Cascades Mountains of Washington has a subalpine forefront characterized by a well-developed terminal moraine, inconspicuous successional moraines, fluting, and outwash. These deposits were depleted of symbiotic fungi when first exposed but colonized by them over time after exposure. Four major groups of plant species in this system are (1) mycorrhiza-independent or facultative mycotrophic, (2) dependent on arbuscular mycorrhizae (AM) (3) dependent on ericoid mycorrhiza (ERM) or ectomycorrhizae (EM), and (4) colonized by dark-septate (DS) endophytes. We hypothesized that availability of mycorrhizal propagules was related to the success of mycorrhiza-dependent plants in colonizing new substrates in naturally evolved ecosystems. To test this hypothesis roots samples of 66 plant species were examined for mycorrhizal colonization. The plants were sampled from communities at increasing distances from the glacier terminus to compare the newest communities with successively older ones. Long established, secondary successional dry meadow communities adjacent to the glacier forefront, and nearby high alpine communities were sampled for comparison. DS were common on most plant species on the forefront. Nonmycorrhizal plants predominated in the earlier successional sites, whereas the proportion of mycorrhizal plants generally increased with age of community. AM were present, mostly at low levels, and nearly absent in two sites of the forefront. ERM were present in all species of Ericaceae sampled, and EM in all species of Pinaceae and Salicaceae. Roots of plants in the long established meadow and heath communities adjacent to the forefront and the high alpine community all had one or another of the colonization types, with DS and AM predominating.     

Seasonal changes in the microbial community of a salt marsh, measured by phospholipid fatty acid analysis

Microbial activity within the environment can have distinct geochemicaleffects, and so changes in a microbial community structure can result ingeochemical change. We examined seasonal changes in both the microbialcommunityand the geochemistry of an inter-tidal salt marsh in north-west England tocharacterise biogeochemical processes occurring at this site.Phospholipid fatty acid (PLFA) analysis of sediment samples collected atmonthly intervals was used to measure seasonal changes in microbial biomass andcommunity structure. The PLFA data were analysed using multivariate techniques(Ward's method and the Mahalanobis distance metric), and we show that the useofthe Mahalanobis distance metric improves the statistical analysis by providingdetailed information on the reasons samples cluster together and identifyingthedistinguishing features between the separate clusters. Five clusters of likesamples were defined, showing differences in the community structure over thecourse of a year.At all times, the microbial community was dominated by PLFA associated withaerobic bacteria, but this was most pronounced in summer (August). Theabundanceof branched fatty acids, a measure of the biomass of anaerobes, started toincrease later in the year than did those associated with aerobes and thefungalbiomarker 18:26 showed a brief late-summer peak.The salt marsh remained mildly oxic throughout the year despite the increase inmicrobial respiration, suggested by the large increases in the abundance ofPLFA, in the warmer months. The conditions therefore remained most favourablefor aerobic species throughout the year, explaining their continual dominanceatthis site. However, as the abundance of PLFA synthesised by anaerobesincreased,increases in dissolved Mn concentrations were observed, which we suggest weredue to anaerobic respiration of Mn(IV) to Mn(II). Overall, the geochemicalconditions were consistent with the microbial community structure and changeswithin it.     

磷脂脂肪酸法在土壤微生物群落分析中的应用

土壤微生物群落的组成一直是土壤学、微生物学和生态学研究的热点问题。我国在这方面的研究处于国际前列,越来越多的研究成果在国际重要刊物上发表。而磷脂脂肪酸(PLFA)法在土壤微生物群落分析中占有举足轻重的地位,国内外学者都热衷于使用该方法。但是PLFA法的使用仍存在一些不足的地方,需要研究学者们慎重使用。本文综述了国际上相关研究,概述了PLFA方法使用的发展历史,应用及挑战。总结了使用和数据解读时需要注意的问题,整理了PLFA法相关的生物标记以及与新方法结合的设想,方便以后研究的开展。     

Changes in microbial nutrient status during secondary succession and its modification by earthworms

Summary Microbial biomass, nutrient (N and P) status, and carbon and nutrient limitation of the microflora were investigated in soils from five different sites (field, 5-, 12-, and about 50-year-old fallow, beechwood), which represent different stages of a secondary succession from a wheat field to the climax ecosystem of a beechwood on limestone. In addition, the effect of faeces production by the substrate feeding earthworm species Octolasion lacteum (Örley) on the nutrient status of the soil microflora of these sites was studied. Humus had accumulated in the soil of the third fallow site, with an enhanced biomass of microflora. However, in the beechwood soil, which had the highest humus content, microbial biomass was lower than in the soil of the third fallow site and similar to that of the field and the two younger fallow sites. In general, soil microbial biomass was little affected by the passage of soil through the gut of O. lacteum. The soil microflora of the field, the 5-, 12-, and about 50-year-old fallow was limited by carbon, whereas in the beechwood soil phosphorus limited microbial growth. NItrogen availability to the soil microflora was low in the two younger fallow sites and high in the field and the third fallow. In the beechwood soil nitrogen supply did not affect microbial carbon utilization. Application of phosphorus stimulated glucose mineralization in the soil of the field, the third fallow, and the beechwood, but not in the two younger fallow sites. Therefor, the nutrient status of the soil microflora seems to have changed during secondary succession: presumably, during the first phase the availability of nitrogen decreased, whereas during the second phase microbial phosphorus supply became more important, which resulted in phosphorus limitation of the soil microflora in the climax ecosystem. The passage of soil through the gut of O. lacteum caused an alteration in the microbial nutrient status. Generally, microbial growth in earthworm casts was limited by carbon. The relative effect of the gut passage of the soils on microbial carbon utilization seems to increase during succession. Therefore, the effect of decomposer invertebrates on microbial nutrient supply seems to increase during secondary succession. In general, nitrogen did not limit microbial carbon utilization in earthworm casts. Phosphorus requirements of the soil microflora were lowered by the gut passage of the soil of the third fallow site and the beechwood, which indicates an increased phosphorus supply in earthworm casts. Howerver, this additional supply was not sufficient to enable optimal carbon utilization by the soil microflora. The results indicate that the effect of decomposer invertebrates on the soil microflora depends on the nutrient status of the ecosystem.     

Plant succession and rhizosphere microbial communities in a recently deglaciated alpine terrain

This study describes how early and late successional plant species affect soil microorganisms in alpine ecosystems. We quantify the relative importance of plant species and soil properties as determinants of belowground microbial communities. Sixteen plant species were selected from six successional stages (4–14–20–43–75–135 years) within the foreland of the Rotmoosferner glacier, Austria, and at one (reference) site outside the foreland. The size, composition and function of the communities of microorganism in the bulk soil and the rhizosphere were characterized by ninhydrin-reactive nitrogen, phospholipid fatty acids and enzyme activities (β-glucosidase, β-xylosidase, N-acetyl-β-glucosaminidase, leucine aminopeptidase, acid phosphatase, sulphatase). The results show that the microbial data could be grouped according to early (up to 43 years) and late-colonizing plant species (75 or more years). In early succession, no plant species or soil age effect was detected on the microbial biomass, phospholipid fatty acids, or enzyme activity. The rhizosphere microbial community was similar to that in the bulk soil, which in turn was determined by the abiotic environmental conditions. In late succession, improved soil conditions probably mediated plant species effects on the belowground microbial community. The most pronounced rhizosphere effects were attributed to plant species of the 75- and 135-year-old sites. The microbial colonization (size, composition, activity) of the bulk soil predominantly followed changes in vegetation cover, plant life forms and soil organic matter. In summary, the observed successional pattern of the above- and belowground communities provides an example of the facilitation models of primary succession.     

Sheep-urine-induced changes in soil microbial community structure

Soil microbial communities play an important role in nutrient cycling and nutrient availability, especially in unimproved soils. In grazed pastures, sheep urine causes local changes in nutrient concentration which may be a source of heterogeneity in microbial community structure. In the present study, we investigated the effects of synthetic urine on soil microbial community structure, using physiological (community level physiological profiling, CLPP), biochemical (phospholipid fatty acid analysis, PLFA) and molecular (denaturing gradient gel electrophoresis, DGGE) fingerprinting methods. PLFA data suggested that synthetic urine treatment had no significant effect on total microbial (total PLFA), total bacterial or fungal biomass; however, significant changes in microbial community structure were observed with both PLFA and DGGE data. PLFA data suggested that synthetic urine induced a shift towards communities with higher concentrations of branched fatty acids. DGGE banding patterns derived from control and treated soils differed, due to a higher proportion of DNA sequences migrating only to the upper regions of the gel in synthetic urine-treated samples. The shifts in community structure measured by PLFA and DGGE were significantly correlated with one another, suggesting that both datasets reflected the same changes in microbial communities. Synthetic urine treatment preferentially stimulated the use of rhizosphere-C in sole-carbon-source utilisation profiles. The changes caused by synthetic urine addition accounted for only 10-15% of the total variability in community structure, suggesting that overall microbial community structure was reasonably stable and that changes were confined to a small proportion of the communities.     

Soil microbial biomass and community composition along an anthropogenic disturbance gradient within a long-leaf pine habitat

Some of the finest surviving natural habitat in the United States is on military reservations where land has been protected from development. However, responsibilities of military training often require disturbance of that habitat. Herein, we show how the soil microbial community of a long-leaf pine ecosystem at Fort Benning, Georgia responds to military traffic disturbances. Using the soil microbial biomass and community composition as ecological indicators, reproducible changes showed increasing traffic disturbance decreases soil viable biomass, biomarkers for microeukaryotes and Gram-negative bacteria, while increasing the proportions of aerobic Gram-positive bacterial and Actinomycete biomarkers. Soil samples were obtained from four levels of military traffic (reference, light, moderate, and heavy) with an additional set of samples taken from previously damaged areas that were remediated via planting of trees and ground cover. Utilizing 17 phospholipid fatty acid (PLFA) variables that differed significantly with land usage, a linear discriminant analysis with cross-validation classified the four groups. Wilks’ lambda for the model was 0.032 (P<0.001). Overall, the correct classifications of profiles was 66% (compared to the chance that 25% would be correctly classified). Using this model, 10 observations taken from the remediated transects were classified. One observation was classified as a reference, three as light trafficked, and six as moderately trafficked. Non-linear artificial neural network (ANN) discriminant analysis was performed using the biomass estimates and all of the 61 PLFA variables. The resulting optimal ANN included five hidden nodes and resulted in an r² of 0.97. The prediction rate of profiles for this model was again 66%, and the 10 observations taken from the remediated transects were classified with four as reference (not impacted), two as moderate, and four as heavily trafficked. Although the ANN included more comprehensive data, it classified eight of the 10 remediated transects at the usage extremes (reference or heavy traffic). Inspection of the novelty indexes from the prediction outputs showed that the input vectors from the remediated transects were very different from the data used to train the ANN. This difference suggests as a soil is remediated it does not escalate through states of succession in the same way as it descends following disturbance. We propose to explore this hysteresis between disturbance and recovery process as a predictor of the resilience of the microbial community to repeated disturbance/recovery cycles.     

Ethanol, BTEX and microbial community interactions in E-blend contaminated soil slurry

Degradation of benzene, toluene, ethylbenzene, m-, p- and o-xylenes (BTEX) and microbial community shifts in soil slurries contaminated with ethanol–gasoline blends (E-blends), containing 10, 50 or 90% (v/v) ethanol (E10, E50 and E90) were studied in soil slurries previously uncontaminated, contaminated by E-blends or ethanol. BTEX originating from E50 degraded fastest whereas from E10 slowest. Among the individual compounds, ethylbenzene degraded fastest (max 30% d⁻¹), and o-xylene slowest (min 1% d⁻¹) during aerobic conditions in previously not contaminated soils. Previous contamination by E-blends increased BTEX degradation significantly (3–19 times) compared with previously uncontaminated soils, whereas previous contamination with ethanol did not show significant difference in BTEX degradation. At least one type of the E-blends during aerobic conditions had a positive effect on total PLFAs (phospholipid fatty acids) and specific PLFAs, i.e. 10Me18:0, 16:1ω6 and cy17:0, but had a negative effect on cy19:0 and 18:2ω6,9c. The effects on total PLFAs, as well as the individual PLFAs, were particularly strong after repeated contamination. The single most affected PLFA was 16:1ω6, which increased 23 times during E10 treatment in soil slurries previously contaminated by E-blends. Altogether, the various E-blends had significantly different effects on BTEX degradation and also on individual PLFAs under aerobic conditions.