Functional diversity as indicator of the recovery of soil health derived from Thlaspi caerulescens growth and metal phytoextraction

Continuous phytoextraction has lately drawn a lot of attention due to its potential for the remediation of metal polluted soils. Although when assessing the success of a phytoextraction process, up till now, emphasis has mostly been placed on metal removal, it is important to highlight that the ultimate objective of a phytoextraction process must be to restore soil health. Consequently, a short-term microcosm study was carried out to evaluate the capacity of an actively growing ecotype of the Zn and Cd hyperaccumulator Thlaspi caerulescens (Lanestosa ecotype) to phytoextract metals from soil and, above all, to assess the potential of soil functional diversity (through the determination of soil enzyme activities and community level physiological profiles) to both determine the toxic effect of metals on soil condition and to monitor the efficiency of a metal phytoextraction process. T. caerulescens plants grown in metal polluted soils showed a shoot metal concentration of 337 mg of Cd, 5670 mg of Zn and 76.6 mg of Pb per kg of dry weight tissue. Apart from confirming its great potential for Zn and Cd phytoextraction, the presence of T. caerulescens, as compared to the metal phytoextraction itself, had the major effect on soil biological parameters. Actually, in metal polluted soils, the presence of T. caerulescens led to a 154, 115, 140, 37 and 164% increase in the activity of β-glucosidase, arylsulphatase, acid phosphatase, alkaline phosphatase and urease, respectively. Metal pollution did not cause a clear inhibition of soil enzyme activities. Contrasting results were obtained with EcoPlates™ versus soil enzyme activities. Actually, the presence of metals led to significantly lower values of Shannon's index calculated from enzyme activities and non-significant higher values of this same index when calculated from EcoPlates™ data. It was concluded that biological indicators of soil health are valid tools to evaluate the success of a metal phytoextraction process.     

Does changing the taxonomical resolution alter the value of soil macroinvertebrates as bioindicators of metal pollution?

Ecological indicators are taxa that are affected by, and indicate effects of, anthropogenic environmental stress or disturbance on ecosystems. There is evidence that some species of soil macrofauna (i.e. diameter >2 mm) constitute valuable biological indicators of certain types of soil perturbations. This study aims to determine which level of taxonomic resolution, (species, family or ecological group) is the best to identify indicator of soil disturbance. Macrofauna were sampled in a set of sites encompassing different land-use systems (e.g. forests, pastures, crops) and different levels of pollution. Indicator taxa were sought using the IndVal index proposed by Dufrêne and Legendre [Dufrêne, M., Legendre, P., 1997. Species assemblages and indicator species: the need for a flexible asymetrical approach. Ecological Monographs 67, 345-366]. This approach is based on a hierarchical typology of sites. The index value changes along the typology and decreases (increases) for generalist (specialist) faunal units (species, families or ecological groups). Of the 327 morphospecies recorded, 19 were significantly associated with a site type or a group of sites (5.8%). Similarly, species were aggregated to form 59 families among which 17 (28.8%) displayed a significant indicator value. Gathering species into 28 broad ecological assemblages led to 14 indicator groups (50%). Beyond the simple proportion of units having significant association with a given level of the site typology, the proportion of specialist and generalist groups changed dramatically when the level of taxonomic resolution was altered. At the species level 84% of the indicator units were specialist, whereas this proportion decreased to 70 and 43% when families and ecological groups were considered. Because specialist groups are the most interesting type of indicators either in terms of conservation or for management purposes we come to the conclusion that the species level is the most accurate taxonomic level in bioindication studies although it requires a high amount of labour and operator knowledge and is time-consuming.     

Structure and function of the soil microbial community in microhabitats of a heavy metal polluted soil

 Particle-size fractionation of a heavy metal polluted soil was performed to study the influence of environmental pollution on microbial community structure, microbial biomass, microbial residues and enzyme activities in microhabitats of a Calcaric Phaeocem. In 1987, the soil was experimentally contaminated with four heavy metal loads: (1) uncontaminated controls; (2) light (300 ppm Zn, 100 ppm Cu, 50 ppm Ni, 50 ppm V and 3 ppm Cd); (3) medium; and (4) heavy pollution (two- and threefold the light load, respectively). After 10 years of exposure, the highest concentrations of microbial ninhydrin-reactive nitrogen were found in the clay (2–0.1 μm) and silt fractions (63–2 μm), and the lowest were found in the coarse sand fraction (2,000–250 μm). The phospholipid fatty acid analyses (PLFA) and denaturing gradient gel electrophoresis (DGGE) separation of 16S rRNA gene fragments revealed that the microbial biomass within the clay fraction was predominantly due to soil bacteria. In contrast, a high percentage of fungal-derived PLFA 18 : 2ω6 was found in the coarse sand fraction. Bacterial residues such as muramic acid accumulated in the finer fractions in relation to fungal residues. The fractions also differed with respect to substrate utilization: Urease was located mainly in the <2 μm fraction, alkaline phosphatase and arylsulfatase in the 2–63 μm fraction, and xylanase activity was equally distributed in all fractions. Heavy metal pollution significantly decreased the concentration of ninhydrin-reactive nitrogen of soil microorganisms in the silt and clay fraction and thus in the bulk soil. Soil enzyme activity was reduced significantly in all fractions subjected to heavy metal pollution in the order arylsulfatase >phosphatase >urease >xylanase. Heavy metal pollution did not markedly change the similarity pattern of the DGGE profiles and amino sugar concentrations. Therefore, microbial biomass and enzyme activities seem to be more sensitive than 16S rRNA gene fragments and microbial amino-sugar-N to heavy metal treatment. Received: 21 January 2000     

Reducing the dimensionality of soil microinvertebrate community datasets using Indicator Species Analysis: Implications for ecosystem monitoring and soil management

Soil microinvertebrates are closely associated with soil decomposition and nutrient cycles and may be particularly responsive indicators for soil management practices. However, identification of appropriate bioindicator species for many systems has been severely limited by a lack of information on species taxonomy, distribution, and functional role. We evaluated Indicator Species Analysis (ISA) as an objective method for assessing the indicator potential of different taxa without regard to their ecological role or expected management response. Restricting ordination and site classification to significant indicator morphotaxa reduced the dimensionality of the community data matrix by 69% while only slightly decreasing the efficiency of unsupervised classification (from 87.2 to 84.4%); the percentage of total variability explained by first two PCA axes increased following ISA. When these same indicator morphotaxa were used to classify an independent set of samples, the percentage of total variability explained by the first two PCA axes increased from 55.3 to 65.2%; cluster analysis of the test dataset correctly classified 47 out of 50 plots by cover type (94% accuracy). However, restriction of analysis to indicator morphotaxa alone reduced detection of differences between sampling dates relative to the complete dataset. Although care needs to be taken to ensure that the dataset used for indicator selection is fully representative of underlying temporal and spatial variability, ISA appears to overcome many of the limitations associated with parametric and multivariate approaches for identifying indicator morphotaxa and has the potential to greatly reduce the taxonomic expertise and labor costs associated with sorting and identification of soil microarthropods.     

污染土壤植物修复技术研究进展

综述了国内外污染土壤植物修复技术研究进展 ,植物修复是利用某些植物对土壤重金属的超量吸收挥发以及对土壤中有机污染物降解等特殊功能 ,并与根际微生物协同作用 ,原位修复污染土壤的方法 ,费用低 ,效果显著 ,环境友好 ,是极具发展潜力的“绿色产业”。     

基于耦合模型的重金属污染土壤植物修复效益空间差异分析

在污染土壤生态修复的同时须兼顾其经济、社会效益,以提高其修复综合效益并推动修复工作的顺利开展。以贵溪市为研究区域,使用耦合模型对污染土地植物修复效益进行空间差异评价并筛选适宜的修复植物类型。研究结果显示:研究区域重金属污染土壤植物修复综合效益存在较大空间差异,其中水泉区重金属污染土壤植物修复效益最低为0.173 6,苏门区重金属污染土壤植物修复效益最高为0.835 4;九牛岗区和庞源区效益相近,效益值小于0.4,属于效益较低区块;在种植植物类型上看,混合种植的树木和乔灌木植物,能产生较好的效益,单纯种植一种植物类型,总体效益低;建议将高效益的苏门区植物类型和种植方式进行推广,并采用红叶石楠和栾树间套种植方式。     

Limitations of soil microbial biomass carbon as an indicator of soil pollution in the field

The size of the soil microbial biomass carbon (SMBC) has been proposed as a sensitive indicator for measuring the adverse effects of contaminants on the soil microbial community. In this study of Australian agricultural systems, we demonstrated that field variability of SMBC measured using the fumigation-extraction procedure limited its use as a robust ecotoxicological endpoint. The SMBC varied up to 4-fold across control samples collected from a single field site, due to small-scale spatial heterogeneity in the soil physicochemical environment. Power analysis revealed that large numbers of replicates (3-93) were required to identify 20% or 50% decreases in the size of the SMBC of contaminated soil samples relative to their uncontaminated control samples at the 0.05% level of statistical significance. We question the value of the routine measurement of SMBC as an ecotoxicological endpoint at the field scale, and suggest more robust and predictive microbiological indicators.     

Changes of microbial properties in (near-) rhizosphere soils after phytoextraction by Sedum alfredii H: A rhizobox approach with an artificial Cd-contaminated soil

The ultimate goal of soil remediation is to restore soil health. Soil microbial parameters are considered to be effective indicators of soil health. The aim of this study was to determine the effects of phytoextraction on microbial properties through the measurement of soil microbial biomass carbon, soil basal respiration and enzyme activities. For this purpose, a pre-stratified rhizobox experiment was conducted with the Cd hyperaccumulator Sedum alfredii H. for phytoextraction Cd from an artificial contaminated soil (15.81 mg kg⁻¹) under greenhouse conditions. The plant and soil samples were collected after growing the plant for three and six months with three replications. The results indicated that the ecotype of S. alfredii H. originating from an ancient silver mining site was a Cd-hyperaccumulator as it showed high tolerance to Cd stress, the shoot Cd concentration were as high as 922.6 mg kg⁻¹ and 581.9 mg kg⁻¹ at the two samplings, and it also showed high BF (58.4 and 36.8 after 3 and 6 months growth), and TF (5.8 and 5.1 after 3 and 6 months growth). The amounts of Cd accumulated in the shoots of S. alfredii reached to an average of 1206 μg plant⁻¹ after 6 months growth. Basal respiration, invertase and acid phosphatase activities of the rhizosphere soil separated by the shaking method were significantly higher (P < 0.01) than that of the near-rhizosphere soil and the unplanted soil after 3 months growth, so were microbial biomass carbon, urease, invertase and acid phosphatase activities of the rhizosphere soil after 6 months growth. Acid phosphatase activity of the 0–2 mm sub-layer rhizosphere soil collected by the pre-stratified method after 3 months growth was significantly higher (P < 0.05) than that of other sub-layer rhizosphere soils and bulk soil, and so were microbial biomass carbon, basal respiration, urease, invertase and acid phosphatase activities of the 0–2 mm sub-layer rhizosphere soil after 6 months growth. It was concluded that phytoextraction by S. alfredii could improve soil microbial properties, especially in rhizosphere, and this plant poses a great potential for the remediation of Cd contaminated soil.     

Soil microbial response during the phytoremediation of a PAH contaminated soil

The aim of this trial was to quantify and compare the responses of soil microbial communities during the phytoremediation of polycyclic aromatic hydrocarbons (PAHs) in a laboratory trial. The experiment was conducted in 1-kg pots and planted treatments consisted of a mixed ryegrass (Lolium perenne) and white clover (Trifolium repens) sward together with a rhizobial inoculum (Rhizobium leguminosarum bv. trifolii). Throughout the 180-d experiment soil microbial biomass and communities of PAH degraders were monitored. PAH degradation was enhanced in planted treatments that received a rhizobial inoculum. Microbial biomass was enhanced in planted treatments, but there were no significant differences between treatments that had received a rhizobial inoculum and those that had not. However, numbers of PAH degraders were greater in the treatment that had received a rhizobial inoculum.     

Suitability of soil microbial parameters as indicators of heavy metal pollution

Several microbial parameters (microbial biomass, respiration, dehydrogenase, phosphatase, sulphatase, glucosidase, protease and urease activities) were measured in soils from five sites located in urban green areas close to roads differing in traffic density. Our aims were to evaluate the suitability of such parameters as field biomarkers of stress induced by heavy metal pollution, and to compare results obtained by single microbial parameters with results given by an index expressing the average microbial (AME) response of the microbial community. Data showed that all parameters were significantly reduced in the sites characterized by the highest load of metals in soil. Dehydrogenase, sulphatase, glucosidase activities and respiration, declined exponentially with increasing metal concentration, whereas phosphatase activity and AME decreased following a sigmoidal type relationship. In contrast, protease, urease and microbial biomass were not significantly correlated with soil metal concentration. Microbial parameters differed both in sensitivity to critical metal concentrations and in the rate of decline at increasing metal loads in soil. Due to the complex interplay of chemical, physical and biological factors which influence microbial activities and biomass, the proposed index (AME) appeared more suitable than single microbial parameters for a biomonitoring study of this type.     

Influences of lignin from paper mill sludge on soil properties and metal accumulation in wheat

The influences of lignin application on soil properties of three different soils, Jiangxi soil (Ultisol, Hapludult), Heilongjiang soil (Alfisol, Entioboralf) and Beijing soil (Alfisol, Haplustalf), and metal accumulation in wheat (Triticum aestivum L.) were studied in a pot experiment. By lignin amendment, soil pH, organic matter (OM) and cation exchange capacity (CEC) increased, except for CEC in the Beijing soil. Analysis showed that available P and K in lignin-amended soils were also elevated, except for P in the Jiangxi soil. A three-step sequential extraction procedure proposed by the Standards, Measurements and Testing Programme (formerly BCR) of the European Commission was used to investigate the fraction redistribution of heavy metals in soils with lignin application. The fractions were specified as B1: water soluble, exchangeable and carbonate bound, and weakly adsorbed; B2: Fe-Mn oxide bound; and B3: organic matter and sulfide bound. Generally, the heavy metal content of the B2 fraction decreased whereas that of the B3 fraction increased. Lignin application to arable soils can not only improve plant growth in vitro, but also reduce the accumulation of the heavy metals Cu, Zn, Cd, Pb, Cr and Ni in wheat plants.     

Effects of heavy metal contamination and remediation on soil microbial communities in the vicinity of a zinc smelter as indicated by analysis of microbial community phospholipid fatty acid profiles

Heavy metal contamination in an area immediately surrounding a zinc smelter has resulted in destruction of over 485 hectares of forest. The elevated levels of heavy metals in these soils have had significant impacts on the population size and overall activity of the soil microbial communities. Remediation of these soils has resulted in increases in indicators of biological activity and viable population size, which suggest recovery of the microbial populations. Questions remain as to how the metal contamination and subsequent remediation at this site have impacted the population structure of the soil microbial communities. In the current study, microbial communities from this site were analyzed by the phospholipid fatty acid (PLFA) procedure. Principal component analysis of the PLFA profiles indicated that there were differences in the profiles for soils with different levels of metal contamination, and that soils with higher levels of metal contamination showed decreases in indicator PLFAs for mycorrhizal fungi, Gram-positive bacteria, fungi, and actinomycetes. PLFA profiles for remediated sites indicated that remediated soils showed increases in indicator PLFAs for fungi, actinomycetes, and Gram-positive bacteria, compared to unremediated metal contaminated soils. These data suggest a change in the population structure of the soil microbial communities resulting from metal contamination and a recovery of several microbial populations resulting from remediation.     

沙漠腹地人工绿地土壤微生物变异与土壤环境因子关系的研究

通过野外采样与室内分析,研究了不同定植年限下塔里木沙漠公路防护林人工绿地土壤微生物的变异规律及其与土壤物理和化学因子间的关系。结果表明:随林龄的增加,土壤微生物数量明显增加,土壤养分含量有所提高,土壤肥力状况明显改善;在土壤微生物的区系组成中,细菌为优势种,占微生物总数的80%以上,放线菌次之,而真菌最少,不到微生物总数的0.1%;土壤微生物数量受土壤环境因子的影响,其中土壤容重、总孔隙度、含水量、有机质和全氮、全磷、全钾及速效氮、速效磷、速效钾含量与细菌、放线菌和真菌的数量均存在极显著相关关系;防护林建设后土壤颗粒逐渐细化。说明防护林的定植促进了咸水灌溉条件下风沙土的发育,土壤质量提高,利于植物的生长,塔里木沙漠公路防护林防风固沙效益明显。     

Stimulated activity of the soil nitrifying community accelerates community adaptation to Zn stress

Field data have shown that soil nitrifying communities gradually adapt to zinc (Zn) after a single contamination event with reported adaptation times exceeding 1 year. It was hypothesized that this relatively slow adaptation relates to the restricted microbial diversity and low growth rate of the soil nitrifying community. This hypothesis was tested experimentally by recording adaptation rates under varying nitrification activities (assumed to affect growth rates) and by monitoring shifts in community composition. Soils were spiked at various Zn concentrations (0-4000 mg Zn kg⁻¹) and two NH₄⁺-N doses (N1, N2) were applied to stimulate growth. A control series receiving no extra NH₄⁺-N was also included. Soils were incubated in pots under field conditions with free drainage. The pore water Zn concentration at which nitrification was halved (EC50, mg Zn l⁻¹) did not change significantly during 12 months in the control series (without NH₄⁺-N applications), although nitrification recovered after 12 months at the highest Zn dose only. The EC50 after 12 months incubation increased by more than a factor 10 with increasing NH₄⁺-N dose (p < 0.05) illustrating that increased activity accelerates adaptation to Zn. Zinc tolerance tests confirmed the role of Zn exposure, time and NH₄⁺-N dose on adaptation. Zinc tolerance development was ascribed to the AOB community since the AOB/AOA ratio (AOB = ammonia oxidizing bacteria; AOA = ammonia oxidizing archaea) increased from 0.4 in the control to 1.4 in the most tolerant community. Moreover, the AOB amoA DGGE profile changed during Zn adaptation whereas the AOA amoA DGGE profile remained unaffected. These data confirm the slow but pronounced adaptation of nitrifiers to Zn contamination. We showed that adaptation to Zn was accelerated at higher activity and was associated with a shift in soil AOB community that gradually dominated the nitrifying community.     

重金属污染矿区复垦土壤微生物生物量及酶活性的研究

对铜矿废弃地复垦土壤微生物生物量及酶活性研究结果表明 ,与对照土壤相比 ,矿区复垦土壤微生物生物量C、N和P均有所降低 ,微生物商Cmic Corg可作为矿区重金属污染土壤微生物学敏感指标之一 ;酶活性变化与此相似 ,脲酶、脱氢酶和酸性磷酸酶与对照差异显著 ,其他则与对照差异明显 ,一定程度削弱了矿区土壤中C、N营养元素周转速率和能量循环     

Surface and subsurface microbial biomass, community structure and metabolic activity as a function of soil depth and season

Microbial biomass, size and community structure along with an estimate of microbial activity and soil chemical parameters were determined at three depths in two soils (e.g. sandy loam Ultic Hapludalf and silt loam Mollic Hapludalf) replicated three times under one winter and summer season. Microbial biomass and community structure were estimated from phospholipid-PO₄ content and fatty acid methyl ester (FAME) measurements. Microbial activity and assimilative capacity were estimated using a ³H-acetate incorporation into phospholipids and by incubating the soil samples at the average winter and summer temperatures, 3 and 20 °C, respectively. We found that the size of the microbial biomass in both the surface and the subsurface soils was not significantly affected by the seasonal variation but activity increased by as much as 83% at the summer temperatures in the surface soil. We demonstrated using FAME analysis that for both soils seasonal changes in the subsurface microbial community occurred. These findings suggest that winter conditions will shift the population activity level in both the surface and subsurface systems and the biochemical structure of the community in the subsurface. In all cases, the inorganic chemical properties of the soil, as a function of season, remained constant. The greatly increased activity of microbial population at the higher temperature will favor the capacity of the system to utilize nutrients or organic materials that may enter soil. During low temperature seasons the capacity of either surface or subsurface soils to assimilate materials is generally diminished but the reduction reflects changes in metabolism and not a reduced biomass size.     

Soil aggregate size mediates the impacts of cropping regimes on soil carbon and microbial communities

Understanding the influence of long-term crop management practices on the soil microbial community is critical for linking soil microbial flora with ecosystem processes such as those involved in soil carbon cycling. In this study, pyrosequencing and a functional gene array (GeoChip 4.0) were used to investigate the shifts in microbial composition and functional gene structure in a medium clay soil subjected to various cropping regimes. Pyrosequencing analysis showed that the community structure (β-diversity) for bacteria and fungi was significantly impacted among different cropping treatments. Functional gene array-based analysis revealed that crop rotation practices changed the structure and abundance of genes involved in C degradation. Significant correlations were observed between the activities of four enzymes involved in soil C degradation and the abundance of genes responsible for the production of respective enzymes, suggesting that a shift in the microbial community may influence soil C dynamics. We further integrated physical, chemical, and molecular techniques (qPCR) to assess relationships between soil C, microbial derived enzymes and soil bacterial community structure at the soil micro-environmental scale (e.g. within different aggregate-size fractions). We observed a dominance of different bacterial phyla within soil microenvironments which was correlated with the amount of C in the soil aggregates suggesting that each aggregate represents a different ecological niche for microbial colonization. Significant effects of aggregate size were found for the activity of enzymes involved in C degradation suggesting that aggregate size distribution influenced C availability. The influence of cropping regimes on microbial and soil C responses declined with decreasing size of soil aggregates and especially with silt and clay micro-aggregates. Our results suggest that long term crop management practices influence the structural and functional potential of soil microbial communities and the impact of crop rotations on soil C turnover varies between different sized soil aggregates. These findings provide a strong framework to determine the impact of management practices on soil C and soil health.     

螯合剂对大叶井口边草Pb、Cd、As吸收性影响研究

用室内土培试验方法,研究在采自田间的Pb、Cd、As、Zn和Cu复合污染土壤上种植大叶井口边草条件下,外源分别添加0、1.5、3、6、12 mmol/kg乙二胺二琥珀酸（EDDS）、氨三乙酸（NTA）和乙二胺四乙酸（EDTA）对大叶井口边草吸收Pb、Cd和As的影响。结果表明,3种螯合剂处理对大叶井口边草生物量没有显著影响,说明大叶井口边草对3种螯合剂耐性较强;6、12 mmol/kg EDTA处理能极显著提高土壤Pb有效态浓度,进而促进大叶井口边草对Pb的吸收。大叶井口边草地上部Pb吸收量最高达（47.4 ± 1.7）mg/kg,是对照的3.66倍。6、12 mmol/kg EDTA处理能极显著地提高土壤中Cd的有效性,但未促进大叶井口边草地上部对Cd吸收。6 mmol/kg EDDS和3 mmol/kg NTA显著提高了土壤中As有效态浓度,进而提高大叶井口边草地上部对As的吸收,大叶井口边草地上部吸收As最高达（276 ± 10） mg/kg。6 mmol/kg EDTA和6 mmol/kg EDDS处理下大叶井口边草提取的Pb、As量最大,分别为（317 ± 53） μg/盆和（873 ± 41）μg/盆,说明6 mmol/kg EDDS处理下大叶井口边草对复合污染土壤中As的修复具有较大的潜力。     

Soil fertility is associated with fungal and bacterial richness,whereas pH is associated with community composition in polar soil microbial communities

Microbial activities in Arctic and Antarctic soils are of particular interest due to uncertainty surrounding the fate of the enormous polar soil organic matter (SOM) pools and the potential to lose unique and vulnerable micro-organisms from these ecosystems. We quantified richness, evenness and taxonomic composition of both fungi and bacteria in 223 Arctic and Antarctic soil samples across 8 locations to test the global applicability of hypotheses concerning edaphic drivers of soil microbial communities that have been primarily developed from studies of bacteria in temperate and tropical systems. We externally validated our model's conclusions with an independent dataset comprising 33 Arctic heath samples. We also explored if our system was responding to large scale climatic or biogeographical processes that we had not measured by evaluating model stability for one location, Mitchell Pennisula, that had been extensively sampled. Soil Fertility (defined as organic matter, nitrogen and chloride content) was the most important edaphic property associated with measures of α-diversity such as microbial richness and evenness (especially for fungi), whereas pH was primarily associated with measures of β-diversity such as phylogenetic structure and diversity (especially for bacteria). Surprisingly, phosphorus emerged as consistently the second most important driver of all facets of microbial community structure for both fungi and bacteria. Despite the clear importance of edaphic factors in controlling microbial communities, our analyses also indicated that fungal/bacterial interactions play a major, but causally unclear, role in structuring the soil microbial communities of which they are a part.     

Soil water availability strongly alters the community composition of soil protists

Drought and heavy rainfall are contrasting conditions expected to result from increasingly extreme weather during climate change; and both scenarios will strongly affect the functioning of soil systems. However, little is known about the specific responses of soil microorganisms, whose functioning is intimately tied to the magnitude of the water-filled pore space in soil. Soil heterotrophic protists, being important aquatic soil organisms are considered as key-regulators of microbial nutrient turnover. We investigated the responses of distinct protist taxa to changes in soil water availability (SWA) using a modified enumeration technique that enabled quantification of protist taxa up to genus level. Our study revealed a non-linear shift of protist abundance with decreasing SWA and this became apparent at a maximum water-filled pore size of ≤40 μm. Generally, taxa containing large specimen were more severely affected by drought, but responses to either drought or rewetting of soils were not uniform among taxa. Changes in water availability may thus affect the functioning of key taxa and soil ecosystems long before aboveground “drought” effects become apparent.