Towards to physiological status of soil microorganisms determined by RNA:dsDNA ratio

Despite soil microorganisms spend most of their lifetime in a state of dormancy, they are quickly activated by substrate input and easily switch to growth. As both the dsDNA- and RNA- contents increase during microbial growth, the RNA:dsDNA ratio reflects a promising predictor, whether the response of a microbial community to environmental changes is due to an increase in population (by dsDNA) or due to an increase in activity (by RNA). This prediction of the RNA:dsDNA ratio can be accomplished by the comparison of microbial incubation approaches with and without addition of easily available substrates. We exhibited the RNA:dsDNA ratios for four contrasting soil types during substrate-induced growth. Overall, after glucose addition, a strong increase of dsDNA and RNA contents were determined in most of the soil types during 72 h of incubation. Furthermore, we identified distinct temporal soil-specific RNA:dsDNA patterns. The dsDNA- and RNA-contents yielded 26–174 and 0.3–30 µg g-1 soil, respectively. The soil texture was strongly associated with the reduction of RNA recovery, by means of an exponential decrease of RNA-content with increasing clay content. The lower RNA recovery in virgin and arable Chernozem (>30%) compared to soil types with lower clay contents (<17% for Retisol, Luvisol and Calcisol) suggests, that the undercount of RNA yields in clayey soils biased the RNA:dsDNA ratio, and subsequently the physiological state of the microbial community is not adequately represented in soils with clay contents exceeding 30%

Organic Nutrients Induced Coupled C- and P-Cycling Enzyme Activities During Microbial Growth in Forest Soils

Besides environmental and soil physical drivers, the functional properties of microbial populations, i. e., growth rate, enzyme production, and maintenance requirements are dependent on the microbes' environment. The soil nutrition status and the quantity and quality of the substrate input, both infer different growth strategies of microorganisms. It is uncertain, how enzyme systems respond during the different phases of microbial growth and retardation in soil. The objective of this study was to uncover the changes of microbial functioning and their related enzyme systems in nutrient-poor and nutrient-rich beech forest soil during the phases of microbial growth. We determined microbial growth via kinetic approach by substrate-induced respiratory response of microorganisms, enabling the estimation of total, and growing biomass of the microbial community. To induce microbial growth we used glucose, while yeast extract simulated additional input of nutrients and factors indicating microbial residues (i.e., necromass compounds). Microbial growth on glucose showed a 12–18 h delay in associated enzyme activity increase or the absence of distinct activity responses (Vmax). β-glucosidase and chitinase (NAG) demonstrated clear differences of Vmax in time and between P-rich and P-poor soils. However, during microbial growth on glucose + yeast extract, the exponential increase in enzymatic activity was clearly stimulated accompanied by a delay of 8–12 h, smoothing the differences in nutrient-acquisition dynamics between the two soils. Furthermore, cross-correlation of β-glucosidase and acid phosphatase between the two sites demonstrated harmonized time constraints, which reflected the establishment of comparable and balanced enzymatic systems within the decomposition network

Microbial hotspots and hot moments in soil: Concept & review

© 2015 Elsevier Ltd. Soils are the most heterogeneous parts of the biosphere, with an extremely high differentiation of properties and processes within nano- to macroscales. The spatial and temporal heterogeneity of input of labile organics by plants creates microbial hotspots over short periods of time - the hot moments. We define microbial hotspots as small soil volumes with much faster process rates and much more intensive interactions compared to the average soil conditions. Such hotspots are found in the rhizosphere, detritusphere, biopores (including drilosphere) and on aggregate surfaces, but hotspots are frequently of mixed origin. Hot moments are short-term events or sequences of events inducing accelerated process rates as compared to the average rates. Thus, hotspots and hot moments are defined by dynamic characteristics, i.e. by process rates.For this hotspot concept we extensively reviewed and examined the localization and size of hotspots, spatial distribution and visualization approaches, transport of labile C to and from hotspots, lifetime and process intensities, with a special focus on process rates and microbial activities. The fraction of active microorganisms in hotspots is 2-20 times higher than in the bulk soil, and their specific activities (i.e. respiration, microbial growth, mineralization potential, enzyme activities, RNA/DNA ratio) may also be much higher. The duration of hot moments in the rhizosphere is limited and is controlled by the length of the input of labile organics. It can last a few hours up to a few days. In the detritusphere, however, the duration of hot moments is regulated by the output - by decomposition rates of litter - and lasts for weeks and months. Hot moments induce succession in microbial communities and intense intra- and interspecific competition affecting C use efficiency, microbial growth and turnover. The faster turnover and lower C use efficiency in hotspots counterbalances the high C inputs, leading to the absence of strong increases in C stocks. Consequently, the intensification of fluxes is much stronger than the increase of pools. Maintenance of stoichiometric ratios by accelerated microbial growth in hotspots requires additional nutrients (e.g. N and P), causing their microbial mining from soil organic matter, i.e. priming effects. Consequently, priming effects are localized in microbial hotspots and are consequences of hot moments. We estimated the contribution of the hotspots to the whole soil profile and suggested that, irrespective of their volume, the hotspots are mainly responsible for the ecologically relevant processes in soil. By this review, we raised the importance of concepts and ecological theory of distribution and functioning of microorganisms in soil

Soil organic matter availability and climate drive latitudinal patterns in bacterial diversity from tropical to cold temperate forests

Bacteria are one of the most abundant and diverse groups of micro-organisms and mediate many critical terrestrial ecosystem processes. Despite the crucial ecological role of bacteria, our understanding of their large-scale biogeography patterns across forests, and the processes that determine these patterns lags significantly behind that of macroorganisms. Here, we evaluated the geographic distributions of bacterial diversity and their driving factors across nine latitudinal forests along a 3,700-km north–south transect in eastern China, using high-throughput 16S rRNA gene sequencing. Four of 32 phyla detected were dominant: Acidobacteria, Actinobacteria, Alphaproteobacteria and Chloroflexi (relative abundance > 5%). Significant increases in bacterial richness and phylogenetic diversity were observed for temperate forests compared with subtropical or tropical forests. The soil organic matter (SOM) mineralisation rate (SOM , an index of SOM availability) explained the largest significant variations in bacterial richness. Variation partition analysis revealed that the bacterial community structure was closely correlated with environmental variables and geographic distance, which together explained 80.5% of community variation. Among all environmental factors, climatic features (MAT and MAP) were the best predictors of the bacterial community structure, whereas soil pH and SOM emerged as the most important edaphic drivers of the bacterial community structure. Plant functional traits (community weighted means of litter N content) and diversity resulted in weak but significant correlations with the bacterial community structure. Our findings provide new evidence of bacterial biogeography patterns from tropical to cold temperate forests. Additionally, the results indicated a close linkage among soil bacterial diversity, climate and SOM decomposition, which is critical for predicting continental-scale responses under future climate change scenarios and promoting sustainable forest ecosystem services. A plain language summary is available for this article. min mi

Spatial distribution and catalytic mechanisms of β-glucosidase activity at the root-soil interface

© 2016 Springer-Verlag Berlin Heidelberg We compared modifications of soil zymography, a new in situ technique to visualize enzyme activities, based on contact of fluorgenic substrate-saturated membranes with soil either through the gel layer (gel zymography) or without gel application (direct zymography). We coupled zymography with quantitative measurements of enzyme kinetics to characterize catalytic mechanisms of β-glucosidase activity at the plant-soil interface including root surface (rhizoplane), rhizosphere, and bulk soil. Direct zymography refined and focused image resolution. The area of hotspots (i.e., spots with most intensive enzyme activity) as well as color intensity ratios estimated using direct zymography exceeded by a factor of 2 the corresponding values obtained with gel zymography. As determined by direct zymography, the percentage of hotspots associated to root surfaces was 58–68 % of total hotspot area. Hotspot area comprised only 6.8 ± 0.1 % of the total area of an image and 9.0 ± 3 % of the root surface area. The intensity of β-glucosidase activity, however, was up to 20 times higher in the hotspots versus bulk soil. The contribution of rhizosphere to β-glucosidase activity of the whole image (77–82 %) was four times higher than the contribution of the root surface. Enzyme kinetic parameters indicated different enzyme systems in bulk and rhizosphere soil. Higher substrate affinity and catalytic efficiency in bulk than in rhizosphere soil suggested relative domination of microorganisms with more efficient enzyme systems in the former. Coupling direct zymography and kinetic assays enabled mapping the two-dimensional (2D) distribution of enzyme activity at the root-soil interface and estimating the catalytic properties of root-associated and soil-associated enzymes

Spatial distribution and catalytic mechanisms of β-glucosidase activity at the root-soil interface

© 2016, Springer-Verlag Berlin Heidelberg.We compared modifications of soil zymography, a new in situ technique to visualize enzyme activities, based on contact of fluorgenic substrate-saturated membranes with soil either through the gel layer (gel zymography) or without gel application (direct zymography). We coupled zymography with quantitative measurements of enzyme kinetics to characterize catalytic mechanisms of β-glucosidase activity at the plant-soil interface including root surface (rhizoplane), rhizosphere, and bulk soil. Direct zymography refined and focused image resolution. The area of hotspots (i.e., spots with most intensive enzyme activity) as well as color intensity ratios estimated using direct zymography exceeded by a factor of 2 the corresponding values obtained with gel zymography. As determined by direct zymography, the percentage of hotspots associated to root surfaces was 58–68 % of total hotspot area. Hotspot area comprised only 6.8 ± 0.1 % of the total area of an image and 9.0 ± 3 % of the root surface area. The intensity of β-glucosidase activity, however, was up to 20 times higher in the hotspots versus bulk soil. The contribution of rhizosphere to β-glucosidase activity of the whole image (77–82 %) was four times higher than the contribution of the root surface. Enzyme kinetic parameters indicated different enzyme systems in bulk and rhizosphere soil. Higher substrate affinity and catalytic efficiency in bulk than in rhizosphere soil suggested relative domination of microorganisms with more efficient enzyme systems in the former. Coupling direct zymography and kinetic assays enabled mapping the two-dimensional (2D) distribution of enzyme activity at the root-soil interface and estimating the catalytic properties of root-associated and soil-associated enzymes

Functional soil organic matter fractions in response to long-term fertilization in upland and paddy systems in South China

Soil organic matter (SOM) and its fractions play key roles in optimizing crop yield and improving soil quality. However, how functional SOM fractions responded to long-term fertilization and their relative importance for C sequestration were less addressed. In this study, we determined the effects of long-term fertilization on six functional SOM fractions (unprotected, physically protected, physico-biochemically protected, physico-chemically protected, chemically protected, and biochemically protected) based on two long-term fertilization experiments carried out in South China. The unprotected coarse particulate organic matter (cPOM), the biochemically and chemically protected silt-sized fractions (NH-dSilt and H-dSilt) were the primary C storage fractions under long-term fertilization, accounting for 23.6–46.2%, 15.7–19.4%, and 14.4–17.4% of the total soil organic carbon (SOC) content in upland soil and 19.5–29.3%, 9.9–15.5%, and 14.2–17.2% of the total SOC content in paddy soil, respectively. Compared with the control treatment (CK) in upland soil, the application of manure combined with mineral NPK (NPKM) resulted in an increase in the SOC content in the cPOM, pure physically protected fraction (iPOM), the physico-chemically protected (H-μSilt), and the chemically protected (H-dSilt) fraction by 233%, 166%, 124%, and 58%, respectively. Besides, the SOC increase in upland soil expressed as SOC content per unit of total SOC for iPOM, H-μSilt, cPOM and H-dSilt were the highest and as large as 283%, 248%, 194%, and 105% respectively. In paddy soil, the highest increase per unit of total SOC was H-dSilt (190%), followed by H-dClay (156%) and H-μSilt (155%). These results suggested that the upland soil could stabilize more C through the pure physical, whereas the chemical protection mechanism played a more important role in paddy soil. Chemical protection mechanism within the microaggregates played important roles in sequestrating C in both upland and paddy soils. Overall, the different responses of functional SOM fractions to long-term fertilization indicate different mechanisms for SOM cycling in terms of C sequestration under upland and paddy systems

Interactive priming effect of labile carbon and crop residues on SOM depends on residue decomposition stage: Three-source partitioning to evaluate mechanisms

© 2018 Elsevier Ltd Inputs of crop residues and labile C (e.g. root exudates) can affect the decomposition rate of soil organic matter (SOM) through the priming effect (PE). Most previous priming studies describe the addition of single labile or residue C, ignoring the interactions of labile C and fresh or decaying crop residues commonly present in field conditions. Using a dual 13C/14C labelling approach in a 62-day incubation, we investigated the effects of adding labile C (40 μg glucose-C g−1 soil) together with wheat shoot or root residues (3.1 mg C g−1 soil) on SOM priming at three residue decomposition stages: intensive (day-1), reduced (day-9) and stabilised (day-24). To estimate the PE, total soil CO2 efflux and microbial biomass were partitioned for three sources: labile C (14C-glucose), plant residues (13C-labelled) and SOM (unlabelled). Without glucose, roots were decomposed less than shoots but induced 1.4-fold stronger cumulative SOM priming (365 μg C g−1 soil) than shoots. Addition of glucose increased SOM priming, with a stronger effect in the presence of shoot than root residues. Glucose addition at the intensive stage of shoots decomposition slightly increased SOM priming. However, compared with residues alone PE, the addition of glucose during reduced residue decomposition stage, increased SOM priming by 60% (roots) to 104% (shoots). Remarkably, this SOM priming after glucose addition was followed by a decline in residue decomposition and by an increase (up to 50%) in SOM-derived C in microbial biomass. Hence, following glucose addition, microorganisms utilised more SOM rather than feeding on decaying residues during reduced decomposition stage. During stabilised residue decomposition stage, the impact of glucose on SOM priming declined again, while the residue decomposition rate remained unaffected. Furthermore, a large proportion of added glucose (up to 10%) was retained in microbial biomass and its mineralisation rate declined strongly (compared with intensive and reduced decomposition stage). Therefore, the glucose amount was not sufficient to influence microbial activities determining SOM or stabilised residue decomposition rates. Overall, SOM decomposition increased by 1- to 4-fold more than the amount of added glucose C, which resulted in a negative net soil C balance compared with residues alone. Thus, we demonstrated for the first time that 1) the interactive effects of glucose (trace amount) and residues on SOM priming depend on plant residue type (higher under shoots than roots) and 2) stage of residues decomposition (higher SOM priming when labile C was added after the end of intensive decomposition stage of plant residues)