Estimation of the total and active microbial biomass in burial mound paleosoils of a different age

Microorganisms that were isolated from steppe soils buried below kurgans from 5800 to 750 years ago were analyzed for the completeness of isolation, total biomass (the sum of glucose-reactivated and resting microbial cells), and active biomass (metabolically active cells). The metabolic state of microbial communities in buried and modern background soils was estimated from the proportion of active and total biomasses. The paleosoils were found to be characterized by lower total and active biomasses and a lower proportion of active microorganisms as compared to the modern background soils. The age-dependent decrease in the content of active microorganisms in the microbial communities of palesoils was not monotonic. For instance, the 4000-year-old paleosoil was characterized by a high total biomass and a relatively low content of active microorganisms, whereas the 1950-year-old paleosoil was characterized by a relatively low total biomass and a relatively high content of active microorganisms. This could reflect the temporal dynamics of paleoclimatic conditions in the geographic region under study.     

The Mechanisms and Time Factor of the Enzyme Structure of a Paleosoil

The data on the enzyme activity of paleosoils of archaeological sites are given. It is shown that the activity of phosphatases and urease in soils of ancient settlements is significantly higher than in modern soils: 1.5–2.0 times for urease and 7–15 times for phosphatase in some cultural layers of the Bronze Age. This is related to a large amount of organic material (garbage, rubbish, excrement, and urea), which entered the soil in ancient times and stimulated soil microorganisms to produce a greater amount of enzymes, whose high activity has been preserved for 4000 years. The location of the enzymes was determined by soil fumigation using chloroform and activation of extracellular enzymes by glycine. The release of intracellular enzymes as a result of fumigation caused a significant increase in phosphatase activity in modern soils and soils of ancient settlements in contrast to the virgin paleosoil of the Bronze Age. The treatment by glycine exerted a smaller effect on the activity of phosphatases, but caused a significant increase in urease activity. This may indicate the predominating extracellular localization of urease in paleosoils of ancient settlements, while phosphatase is characterized by both extra- and intracellular localization.

     

Reactivation of dormant and nonculturable bacterial forms from paleosoils and subsoil permafrost

Methods of reactivating the dormant forms (DFs) and nonculturable cells (NCs) of the bacterial communities of buried paleosoils and subsoil permafrost stored for long periods of time (thousands to millions of years), including completely sterile samples (CFU = 0), were developed. They were based on washing the DFs and NCs to remove anabiosis autoinducers (spore germination autoinhibitors) and introducing low molecular weight extracellular growth regulators of microbial or plant origin, such as alkylhydroxybenzenes of the alkylresorcinol subtype, indoleacetic acid, and wheat germ agglutinin. It was revealed that the dormant communities of permafrost and buried soils differed in their sensitivity to reactivating factors, probably due to different natural storage conditions of the tested soil substrates and the heterogeneity of dormant populations. The latter was confirmed by FISH (fluorescent in situ hybridization): applying the reactivation methods to the cells of the dormant permafrost community resulted in an increase in the number of metabolically active cells from 5 to 77% of their total number. In contrast, the addition of microbial anabiosis autoinducers (C₁₂-AHB) to background surface soil and permafrost samples caused the transition of bacterial cells to the dormant or the nonculturable state, depending on the C₁₂-AHB concentration and the sensitivity of the cells from the control soil or permafrost’ to it. The data obtained contribute to our knowledge concerning the role of intercellular communication factors and the survival of microorganisms under extreme environmental conditions.     

The effect of pyrogenically modified substrates on mineralizing activity and growth strategies of microorganisms of grey forest soil

The rates of the mineralization processes initiated by the input of plant residues and pyrogenically modified plant material into gray forest soil under forests and meadows were assayed. While meadow plant residues was mineralized more rapidly than the forest floor, decomposition of the pyrogenic material resulted in disproportional changes in CO₂ emission from soils. Statistical treatment showed that the respiratory activity of CO₂ emission by heterotrophic microorganisms, which is a physiological characteristic of microbial communities, is 89% determined by the substrate quality. The maximal specific growth rate, which reflects the functional changes in microbial communities, was affected by the cenosis (36%) and the substrate (30%). Most of the carbon of the original plant material (up to 90%) was removed during the burning of plant substrates. The remaining compounds in the pyrogenically transformed material changed the process of mineralization in soil compared both to the control variant and to soil enriched with plant residues. Input of plant residues and ash into the soil resulted in increased total and active biomass, while the maximal specific growth rate decreased and the generation time for the active biomass increased. In the case of soils with plant residues, these changes in the state of microbial communities were brief and occurred during the period of intense mineralization (0–5 days), while, in soils with plant ash, stable changes were revealed after more prolonged incubation. Experimental determination of the microbial biomass turnover time (MTT) by means of two methods (from the ratio between the microbial biomass and respiration and from microbial specific growth rates) made it possible to determine the economical coefficient Y for microbial communities metabolizing the substrates of different availability. Depending on the experimental variant, the Y values varied from 0.22 to 0.51. Decreased maximal specific growth rate and increased values of Y (the coefficient of efficiency of substrate utilization) showed the predominant contribution of K-strategists in the mineralization of low available substrates in soil. The balance calculations and physiological characteristics of the microbial community suggested that the priming effect was most probable in soils enriched with plant ash.     

Influence of corn,switchgrass, and prairie cropping systems on soil microbial communities in the upper Midwest of the United States

Because soil microbes drive many of the processes underpinning ecosystem services provided by soils, understanding how cropping systems affect soil microbial communities is important for productive and sustainable management. We characterized and compared soil microbial communities under restored prairie and three potential cellulosic biomass crops (corn, switchgrass, and mixed prairie grasses) in two spatial experimental designs – side‐by‐side plots where plant communities were in their second year since establishment (i.e., intensive sites) and regionally distributed fields where plant communities had been in place for at least 10 years (i.e., extensive sites). We assessed microbial community structure and composition using lipid analysis, pyrosequencing of rRNA genes (targeting fungi, bacteria, archaea, and lower eukaryotes), and targeted metagenomics of nifH genes. For the more recently established intensive sites, soil type was more important than plant community in determining microbial community structure, while plant community was the more important driver of soil microbial communities for the older extensive sites where microbial communities under corn were clearly differentiated from those under switchgrass and restored prairie. Bacterial and fungal biomasses, especially biomass of arbuscular mycorrhizal fungi, were higher under perennial grasses and restored prairie, suggesting a more active carbon pool and greater microbial processing potential, which should be beneficial for plant acquisition and ecosystem retention of carbon, water, and nutrients.     

Soil microbial carbon uptake characteristics in relation to soil management

Abstract The kinetics of glucose uptake by soil microbial communities in 16 different soild (7 under monocultures and 9 under crop rotations) differing in microbial biomass content, % C_org, pH and clay content were investigated at 22°C. The V _max value of microbial bimasses under monoculture, was o.27 μg C_gluc · μg⁻¹ C_mic · h⁻¹ (range 0.18–0.44), twice as high as the mean value of V _max of microbial biomasses under rotations (0.13 μg C_gluc, range 0.07–0.19). Mean values of K _m were 714 μg C_gluc and 290 μg C_gluc · g⁻¹ soil, respectively.
These differences were highly significant ( P =0.001, based on SE) and could not be relate to particle size distribution of the soils, pH or C_org. A Michaelis-Menten type uptake response was apparent over the total range of glucose concentrations used (45.4–1453.3 μg C_gluc · g⁻¹ soil) for microbial biomasses under rotation while the majority of microbial biomasses under monocultures showed a similar response only at low glucose concentrations. A different uptake mechanism appeared to be involved at higher glucose concentrations (similar to diffusion) in monoculture soils.     

Microbial biomass and growth kinetics of microorganisms in chernozem soils under different farm land use modes

The carbon content of microbial biomass and the kinetic characteristics of microbial respiration response to substrate introduction have been estimated for chernozem soils of different farm lands: arable lands used for 10, 46, and 76 years, mowed fallow land, non-mowed fallow land, and woodland. Microbial biomass and the content of microbial carbon in humus (Cmic/Corg) decreased in the following order: soils under forest cenoses-mowed fallow land-10-year arable land-46- and 75-year arable land. The amount of microbial carbon in the long-plowed horizon was 40% of its content in the upper horizon of non-mowed fallow land. Arable soils were characterized by a lower metabolic diversity of microbial community and by the highest portion of microorganisms able to grow directly on glucose introduced into soil. The effects of different scenarios of carbon sequestration in soil on the reserves and activity of microbial biomass are discussed.     

Dynamics of Organic Matter Decomposition and Microflora Composition of Forest Litter in Artificial Biogeocenoses

The dynamics of litter stock, microbial biomass, and composition and structure of microbial communities, were studied in the course of soil organic matter transformation during vegetation season. The dynamics of litter stock in coniferous and deciduous forests proved to correlate with the biomass and total abundance of microorganisms, particularly, with the proportion of microfungi in the microbial community.     

Soil Microbial Communities of Eastern Antarctica

Investigation of microbial communities of Antarctica soils is a very important field of research that expands our knowledge of microbial participation in primary soil formation and specific features of their communities in extreme habitats, and it is of considerable interest in directed search of for microorganisms as potential biotechnological objects. The results of long-term (2012–2017) complex studies on soil microbial communities of the Russian East Antarctica polar stations at Shirmakher oasis (Novolazarevskaya station), the Larsemann Hills (Progress station), and the Tala Hills (Molodezhnaya station) are presented in this review. The assessment of biomass of soil microorganisms by the methods of direct microscopy has been carried out for the first time for this region. The general amount of microbial biomass is small; the fungi dominate (77–99%). The unique features of Antarctic soils are the high content and morphological diversity of small forms of microorganisms: fungi are presented by mainly single-celled structures (small spores and yeasts), while bacteria by ultrafine (filtering) forms. At the same time, microorganisms can significantly contribute to such important ecological functions of soil as the emission of greenhouse gases, especially during the warm season with the stable positive temperatures of the soil. This should be considered during creation of models and forecasts of global warming. The use of various isolation techniques for the analysis of the soil microbial population, together with the succession approach, significantly expand the information about taxonomic diversity of cultivated fungi and bacteria in Antarctica soils.     

Microbial biomass and growth kinetics of microorganisms in chernozem soils under different land use modes

The carbon content of microbial biomass and the kinetic characteristics of microbial respiration response to substrate addition have been estimated for chernozem soils under different land use: arable lands used for 10, 46, and 76 years, mowed meadow, natural forest, and forest shelter belt. Microbial biomass and the content of microbial carbon in humus (C_mic /C_org) decreased in the following order: soils under forest cenoses—mowed meadow—10-year arable land—46- and 75-year arable land. The amount of microbial carbon in the long-plowed horizon was 40% of its content in the upper horizon of natural forest. Arable soils were characterized by a lower metabolic diversity of microbial community and by the highest portion of microorganisms able to grow directly on glucose introduced into soil. The effects of different scenarios of carbon sequestration in soil on the amounts and activity of microbial biomass are discussed.     

Molecular and Functional Assessment of Bacterial Community Convergence in Metal-Amended Soils

Species diversity and the structure of microbial communities in soils are thought to be a function of the cumulative selective pressures within the local environment. Shifts in microbial community structure, as a result of metal stress, may have lasting negative effects on soil ecosystem dynamics if critical microbial community functions are compromised. Three soils in the vicinity of a copper smelter, previously contaminated with background, low and high levels of aerially deposited metals, were amended with metal-salts to determine the potential for metal contamination to shape the structural and functional diversity of microbial communities in soils. We hypothesized that the microbial communities native to the three soils would initially be unique to each site, but would converge on a microbial community with similar structure and function, as a result of metal stress. Initially, the three different sites supported microbial communities with unique structural and functional diversity, and the nonimpacted site supported inherently higher levels of microbial activity and biomass, relative to the metal-contaminated sites. Amendment of the soils with metal-salts resulted in a decrease in microbial activity and biomass, as well as shifts in microbial community structure and function at each site. Soil microbial communities from each site were also observed to be sensitive to changes in soil pH as a result of metal-salt amendment; however, the magnitude of these pH-associated effects varied between soils. Microbial communities from each site did not converge on a structurally or functionally similar community following metal-salt amendment, indicating that other factors may be equally important in shaping microbial communities in soils. Among these factors, soil physiochemical parameters like organic matter and soil pH, which can both influence the bioavailability and toxicity of metals in soils, may be critical.     

Temperature as an autoecological factor of chitinolytic microbial complex formation in soils

The dynamics of carbon dioxide emission from soil was studied during chitinolytic succession induced by humidification and chitin introduction at different temperatures (5, 27, and 50°C) using gas chromatography. The abundance and biomass of the chitinolytic bacterial and actinomycete complex in soil were evaluated by luminescent microscopy. Active development of the chitinolytic microbial complexes was observed at all studied temperatures. The most active growth of chitinolytic microorganisms was observed at high temperature during early succession and at low temperature during late succession. High and low temperatures provided for active development of the chitinolytic microbial complex in soils confined to warm climatic zones (brown desert-steppe soil) and soils of temporary zones (gray forest soil). Actinomycetes demonstrated the most active growth among chitinolytic microorganisms in the studied soil samples both at low and high temperatures.     

Metabolic Status of Bacteria and Fungi in the Rhizosphere of Ponderosa Pine Seedlings

We determined the quantity and metabolic status of bacteria and fungi in rhizosphere and nonrhizosphere soil from microcosms containing ponderosa pine seedlings. Rhizosphere soil was sampled adjacent to coarse, fine, or young roots. The biovolume and metabolic status of bacterial and fungal cells was determined microscopically and converted to total and active biomass values. Cells were considered active if they possessed the ability to reduce the artificial electron acceptor 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloride (INT) to visible intracellular deposits of INT formazan. A colorimetric assay of INT formazan production was also used to assess dehydrogenase activity. INT-active microorganisms made up 44 to 55% of the microbial biomass in the soils studied. The proportion of fungal biomass that exhibited INT-reducing activity (40 to 50%) was higher than previous estimates of the active proportion of soil fungi determined by using fluorescein diacetate. Comparison between soils from different root zones revealed that the highest total and INT-active fungal biomass was adjacent to fine mycorrhizal roots, whereas the highest total and active bacterial biomass was adjacent to the young growing root tips. These observations suggest that fungi are enhanced adjacent to the fine roots compared with the nonrhizosphere soil, whereas bacteria are more responsive than fungi to labile carbon inputs in the young root zone. Colorimetric dehydrogenase assays detected gross differences between bulk and rhizosphere soil activity but were unable to detect more subtle differences due to root types. Determination of total and INT-active biomass has increased our understanding of the role of spatial compartmentalization of bacteria and fungi in rhizosphere carbon flow.     

Relationship of Atmospheric Pollution Characterized by Gas (NO2) and Particles (PM10) to Microbial Communities Living in Bryophytes at Three Differently Polluted Sites (Rural, Urban, and Industrial)

Atmospheric pollution has become a major problem for modern societies owing to its fatal effects on both human health and ecosystems. We studied the relationships of nitrogen dioxide atmospheric pollution and metal trace elements contained in atmospheric particles which were accumulated in bryophytes to microbial communities of bryophytes at three differently polluted sites in France (rural, urban, and industrial) over an 8-month period. The analysis of bryophytes showed an accumulation of Cr and Fe at the rural site; Cr, Fe, Zn, Cu, Al, and Pb at the urban site; and Fe, Cr, Pb, Al, Sr, Cu, and Zn at the industrial site. During this study, the structure of the microbial communities which is characterized by biomasses of microbial groups evolved differently according to the site. Microalgae, bacteria, rotifers, and testate amoebae biomasses were significantly higher in the rural site. Cyanobacteria biomass was significantly higher at the industrial site. Fungal and ciliate biomasses were significantly higher at the urban and industrial sites for the winter period and higher at the rural site for the spring period. The redundancy analysis showed that the physico-chemical variables ([NO₂], relative humidity, temperature, and site) and the trace elements which were accumulated in bryophytes ([Cu], [Sr], [Pb]) explained 69.3% of the variance in the microbial community data. Moreover, our results suggest that microbial communities are potential biomonitors of atmospheric pollution. Further research is needed to understand the causal relationship underlined by the observed patterns.     

Effects of salinity on microbial tolerance to drying and rewetting

Soil salinity and fluctuations in soil matric potential are stressors for soil microorganisms which, in turn, may affect soil organic matter turnover. In response to salinity and low soil water content, many microorganisms accumulate osmolytes. Therefore, it is conceivable that microorganisms in saline soils are more tolerant to drying and rewetting (DRW) stress than those in non-saline soils. An experiment was carried out with three different salinity levels: electrical conductivity (EC_1:5) 0, 2 and 4 dS m^?1 (EC0, EC2, EC4), and two water treatments: a constantly moist control or two DRW cycles. Respiration as an indicator of microbial activity was measured throughout the 59 days of incubation. At the end of the second dry period (day 35) and at the end of the following moist incubation (day 59), microbial biomass and microbial community structure were determined by phospholipid fatty acid (PLFA) analysis. Increasing salinity decreased microbial activity but did not affect its resistance to DRW. On day 59, cumulative respiration decreased in the order EC0 > EC2 > EC4 with no differences between water treatments. Fungal biomass was negatively affected by salinity at the end of the experiment, while bacterial biomass was unaffected. Microbial community structure in moist treatments differed between salinity levels, with EC4 influencing microbial community structure earlier than EC2. The resistance of microbial communities to DRW stress was salt level dependent; only beyond a critical salinity level adaptation to salt stress was able to reduce the impact of water stress on microbial community structure.     

不同肥力棕壤全氮和微生物量氮对外源玉米残体氮的响应

以棕壤玉米长期连作定位试验(27a)形成的高低两种肥力水平土壤为研究对象,采用~(15)N标记的玉米植株为试验试材,分别向两种土壤中加入玉米根、茎、叶(共8个处理),采用室内模拟培养与~(15)N同位素示踪技术,旨在弄清玉米根、茎、叶添加后不同肥力土壤全氮含量及微生物量氮的变化规律。结果表明:(1)添加玉米根、茎、叶后低肥力棕壤全氮含量提升幅度分别比高肥力棕壤高5.75%、4.77%和3.75%,外源新氮的贡献率分别比高肥力棕壤高3.54%、3.28%和2.49%,说明不同肥力土壤对玉米残体添加的响应程度不同,低肥力棕壤对外源新氮施入后的响应更敏感,固定能力更强。(2)在添加玉米残体的56d培养时间内,低肥力棕壤中微生物量氮平均增加0.83—0.98倍,高肥力棕壤中微生物量氮平均增加0.87—1.56倍,可以看出不同部位玉米植株添加后均能显著促进土壤微生物量氮的积累,说明外源有机物输入是刺激土壤微生物数量和活性的重要因素,并且在高肥力土壤中刺激作用更加显著。此外,高肥力土壤添加茎和叶处理微生物量氮显著高于根添加处理,但低肥力土壤中根、茎和叶添加处理土壤中微生物量氮之间无显著差异。外源有机氮输入对土壤氮库的贡献与土壤的肥力水平及不同残体部位自身的物质组成特性密切相关。     

不同覆土基质微生物结构特征研究及其对双孢蘑菇产量的影响

覆土是影响双孢蘑菇产量、质量和出菇整齐度的重要因子,利用现代分子生态学的方法快速、准确地对不同覆土基质微生物结构特征进行检测,以进一步了解微生物群落与双孢蘑菇相互作用关系。测定了不同覆土的理化特性,应用PCR技术对不同覆土材料提取总DNA,扩增细菌16S rDNA和真菌28S rDNA,运用变性梯度凝胶电泳技术对PCR产物进行分析,研究双孢蘑菇不同覆土基质微生物结构特征。结果表明：不同处理的覆土材料微生物群落的基因具有多样性,其中细菌群落基因多样性存在差异,使用纯泥炭与粉碎稻草处理差异最大,相似性仅为62%;通过真菌28S rDNA变性梯度凝胶电泳结果显示,粉碎稻草处理多样性指数最高,达3.576,但随着泥炭比例的提高,覆土处理中真菌群落的多样性相对减少;栽培试验发现,双孢蘑菇子实体形成量、总产量可能与覆土中的真菌群落多样性呈负相关。     

Advanced mine restoration protocols facilitate early recovery of soil microbial biomass,activity and functional diversity

Many ecosystem restoration programmes can take over 15 years to achieve ecosystem functioning comparable to that of an unmodified ecosystem, therefore a reliable shorter-term method of assessing and monitoring ecosystem recovery is needed to ensure that recovery is following an appropriate trajectory. Soil microbes respond to environmental change relatively quickly, and shifts in microbial communities can reflect the current status of their environment. As well as potentially acting as ‘indicator communities’, microbes play an integral role in restoring ecosystem functions. On an active opencast mine on New Zealand's West Coast, three main restoration methods are used, differing in cost and restoration effort. They range from most expensive (1) vegetation direct transfer (VDT), to (2) biosolids-amended stockpiles that are spread and replanted, and (3) untreated stockpiles that are spread and replanted. We assessed the impacts of these methods on soil microbial communities by measuring microbial biomass, dehydrogenase activity, community level physiological profile (CLPP) and functional diversity as measured by carbon substrate utilisation, where restored sites were 5 years old or less. These measures were compared to an unmodified reference ecosystem in the same location. Microbial activity and biomass were highest in pristine habitats, followed by VDT and biosolids-amended soils, then untreated stockpile soil. When compared to all other treatments untreated stockpiled soils had significantly different CLPPs and significantly reduced microbial biomass and activity; microbial biomass was an order of magnitude lower than in pristine soils. Functional diversity and richness did not differ between pristine, VDT and biosolids-amended soils, but were higher than in untreated stockpiled soils. CLPPs did not differ between pristine habitat soil and VDT soil but biosolids-amended and untreated stockpiled soils were significantly different to pristine soil. This study has shown that soil microbial communities are a valuable tool to assess restoration progress, and that ecosystem restoration can begin in a relatively short time following investment in appropriate restoration strategy, ultimately benefiting recolonisation by plants and animals.     

Biogeographical patterns of soil molecular microbial biomass as influenced by soil characteristics and management

Aim The spatial organization of soil microbial communities on large scales and the identification of environmental factors structuring their distribution have been little investigated. The overall objective of this study was to determine the spatial patterning of microbial biomass in soils over a wide extent and to rank the environmental filters most influencing this distribution. Location French territory using the French Soil Quality Monitoring Network. This network covers the entire French territory and soils were sampled at 2150 sites along a systematic grid. Methods The soil DNA extracted from all these soils was expressed in terms of soil molecular microbial biomass and related to other soil and land‐use data over French territory. Results This study provides the first extensive map of microbial biomass and reveals the heterogeneous and spatially structured distribution of this biomass on the scale of France. The main factors driving biomass distribution are the physico‐chemical properties of the soil (texture, pH and total organic carbon) as well as land use. Soils from land used for intensive agriculture, especially monoculture and vineyards, exhibited the smallest biomass pools. Interestingly, factors known to influence the large‐scale distribution of macroorganisms, such as climatic factors, were not identified as important drivers for microbial communities. Main conclusions Microbial abundance is spatially structured and dependent on local filters such as soil characteristics and land use but is relatively independent of global filters such as climatic factors or the presence of natural barriers. Our study confirms that the biogeography of microorganisms differs fundamentally from the biogeography of ‘macroorganisms’ and that soil management can have significant large‐scale effects.     

西南峡谷型喀斯特坡地土壤微生物量C、N、P空间变异特征

土壤微生物是陆地生态系统中最活跃的成分,它推动着生态系统的能量和物质循环,被公认为土壤生态系统变化的预警及敏感指标。以西南峡谷型喀斯特坡地为研究对象,基于网格法取样,结合经典统计学和地统计学方法,揭示了土壤微生物生物量的空间分布与格局及其主要影响因子。结果表明,西南峡谷型喀斯特坡地土壤微生物生物量碳(MBC)、氮(MBN)、磷(MBP)、碳氮比(MBC/MBN)、碳磷比(MBC/MBP)适宜,MBC、MBN、MBP变异均很大;空间自相关性明显,除MBP最佳拟合模型为球状模型外,其他指标均为指数模型。C0/(C0+C)均25%(4.9%—6.2%),呈强烈的空间相关,这主要由结构性变异引起。Kriging等值线图表明,MBC、MBN的高值区集中在坡中上部;MBP的格局明显不同,高值区集中在坡脚;MBC/MBN斑块较大,变化缓和;MBC/MBP的空间分布规律不明显,斑块多而破碎。西南峡谷型喀斯特坡地土壤微生物量空间分布的影响因子很多,其中,影响土壤微生物量碳和氮的主要因子有土层厚度、pH、碱解氮。西南峡谷型喀斯特坡地土壤微生物不仅存在着小尺度的空间分布格局,而且不同土壤微生物属性的空间分布不同。因此,应采取适宜措施,激活土壤微生物活性。