Carbon Storage Regulator Research Articles

Expression, regulation and physiological roles of the five Rsm proteins in Pseudomonas syringae pv. tomato DC3000

Proteins belonging to the RsmA (regulator of secondary metabolism)/CsrA (carbon storage regulator) family are small RNA-binding proteins that play crucial roles post-transcriptionally regulating gene expression in many Gram-negative and some Gram-positive bacteria. Although most of the bacteria studied have a single RsmA/CsrA gene, Pseudomonas syringae pv. tomato (Pto) DC3000 encodes five Rsm proteins: RsmA/CsrA2, RsmC/CsrA1, RsmD/CsrA4, RsmE/CsrA3, and RsmH/CsrA5. This work aims to provide a comprehensive analysis of the expression of these five rsm protein-encoding genes, elucidate the regulatory mechanisms governing their expression, as well as the physiological relevance of each variant. To achieve this, we examined the expression of rsmA, rsmE, rsmC, rsmD, and rsmH within their genetic contexts, identified their promoter regions, and assessed the impact of both their deletion and overexpression on various Pto DC3000 phenotypes. A novel finding is that rsmA and rsmC are part of an operon with the upstream genes, whereas rsmH seems to be co-transcribed with two downstream genes. We also observed significant variability in expression levels and RpoS dependence among the five rsm paralogs. Thus, despite the extensive repertoire of rsm genes in Pto DC3000, only rsmA, rsmE and rsmH were significantly expressed under all tested conditions (swarming, minimal and T3SS-inducing liquid media). Among these, RsmE and RsmA were corroborated as the most important paralogs at the functional level, whereas RsmH played a minor role in regulating free life and plant-associated phenotypes. Conversely, RsmC and RsmD did not seem to be functional under the conditions tested.

Evolution recovers the fitness of Acinetobacter baylyi strains with large deletions through mutations in deletion-specific targets and global post-transcriptional regulators.

Organelles and endosymbionts have naturally evolved dramatically reduced genome sizes compared to their free-living ancestors. Synthetic biologists have purposefully engineered streamlined microbial genomes to create more efficient cellular chassis and define the minimal components of cellular life. During natural or engineered genome streamlining, deletion of many non-essential genes in combination often reduces bacterial fitness for idiosyncratic or unknown reasons. We investigated how and to what extent laboratory evolution could overcome these defects in six variants of the transposon-free Acinetobacter baylyi strain ADP1-ISx that each had a deletion of a different 22- to 42-kilobase region and two strains with larger deletions of 70 and 293 kilobases. We evolved replicate populations of ADP1-ISx and each deletion strain for ~300 generations in a chemically defined minimal medium or a complex medium and sequenced the genomes of endpoint clonal isolates. Fitness increased in all cases that were examined except for two ancestors that each failed to improve in one of the two environments. Mutations affecting nine protein-coding genes and two small RNAs were significantly associated with one of the two environments or with certain deletion ancestors. The global post-transcriptional regulators rnd (ribonuclease D), csrA (RNA-binding carbon storage regulator), and hfq (RNA-binding protein and chaperone) were frequently mutated across all strains, though the incidence and effects of these mutations on gene function and bacterial fitness varied with the ancestral deletion and evolution environment. Mutations in this regulatory network likely compensate for how an earlier deletion of a transposon in the ADP1-ISx ancestor of all the deletion strains restored csrA function. More generally, our results demonstrate that fitness lost during genome streamlining can usually be regained rapidly through laboratory evolution and that recovery tends to occur through a combination of deletion-specific compensation and global regulatory adjustments.

Successional stages in Mediterranean grasslands differ in the quality of ecosystem services in urban greenspaces

Urban habitats represent new opportunities to study natural processes such as ecological succession. Our work focused for the first time on the changes in ecosystem services occurring in the ecological succession from annual to perennial grasslands in Mediterranean peri-urban greenspaces. Student’s t test was used to analyse the influence of the grassland type on plant and soil features, and showed that soil under perennial grassland had a significantly higher C content and arylamidase and arylsulfatase activity than annual grasslands. In contrast, annual grasslands showed a significantly higher plant-species richness, and their soils had higher bulk density and phenoloxidase and arylamidase activity compared to perennial grasslands. Neither the labile nor recalcitrant fractions of the organic matter showed any significant difference between communities. When all the soil factors were included together, Redundancy Analysis revealed a significant gradient in soil phenoloxidase activity and organic matter distinguishing perennial from annual grasslands. We conclude that when annual grasslands give way to perennial grasslands through natural succession in Mediterranean greenspaces, certain ecosystem services such as soil carbon storage and water regulation improve, while biodiversity maintenance declines. Thus, Mediterranean urban greenspaces where natural succession occurs should be handled in such a way as to preserve both type of habitats in order to improve a wider range of ecosystem services.

Scaling carbon removal systems: deploying direct air capture amidst Canada’s low-carbon transition

Carbon dioxide removal (CDR) technologies, such as direct air carbon capture and storage (DACCS), will be critical in limiting the rise of the average global temperature over the next century. Scaling up DACCS technologies requires the support of a complex array of policies and infrastructure across multiple overlapping policy areas, such as climate, energy, technology innovation and resource management. While the literature on DACCS and other CDR technologies acknowledges the path-dependent nature of policy development, it has tended to focus on abstract policy prescriptions that are not rooted in the specific political, social and physical (infrastructural) context of the implementing state. To address this gap, this paper provides a country-level study of the emerging DACCS policy regime in Canada. Drawing on the existing literature that identifies idealized (acontextual) policy objectives that support DACCS development and effective regulation, we identify the actionable policy objectives across six issue domains: general climate mitigation strategies; energy and resource constraints; carbon storage and transport regulation and infrastructure; financing scale-up and supporting innovation; removal and capture technology availability and regulation; and addressing social acceptability and public interest. Using a database of Canadian climate policies (n = 457), we identify policies within the Canadian (federal and provincial) policy environment that map to the idealized policy objectives within each of these domains. This exercise allows us to analyze how key policy objectives for DACCS development are represented within the Canadian system, and enables us to identify potential niches, and landscape influences within the system, as well as gaps and potential barriers to the system transition process. This paper contributes to our understanding of national DACCS policy development by providing a framework for identifying components of the DAC system and linking those components to desired policy outcomes and may provide a basis for future cross-country comparisons of national-level DACCS policy.

Peeling the onion: additional layers of regulation in the acid stress response.

Bacteria are capable of withstanding large changes in osmolality and cytoplasmic pH, unlike eukaryotes that tightly regulate their pH and cellular composition. Previous studies on the bacterial acid stress response described a rapid, brief acidification, followed by immediate recovery. More recent experiments with better pH probes have imaged single living cells, and we now appreciate that following acid stress, bacteria maintain an acidic cytoplasm for as long as the stress remains. This acidification enables pathogens to sense a host environment and turn on their virulence programs, for example, enabling survival and replication within acidic vacuoles. Single-cell analysis identified an intracellular pH threshold of ~6.5. Acid stress reduces the internal pH below this threshold, triggering the assembly of a type III secretion system in Salmonella and the secretion of virulence factors in the host. These pathways are significant because preventing intracellular acidification of Salmonella renders it avirulent, suggesting that acid stress pathways represent a potential therapeutic target. Although we refer to the acid stress response as singular, it is actually a complex response that involves numerous two-component signaling systems, several amino acid decarboxylation systems, as well as cellular buffering systems and electron transport chain components, among others. In a recent paper in the Journal of Bacteriology, M. G. Gorelik, H. Yakhnin, A. Pannuri, A. C. Walker, C. Pourciau, D. Czyz, T. Romeo, and P. Babitzke (J Bacteriol 206:e00354-23, 2024, https://doi.org/10.1128/jb.00354-23) describe a new connection linking the carbon storage regulator CsrA to the acid stress response, highlighting new additional layers of complexity.

Residue Kinetics of Insecticide Pymetrozine in Rice Field Soil

Soils are the most diverse and complex ecosystem in the world. In addition to providing humanity with 98.8% of its food, soils provide a broad range of other services, from carbon storage and greenhouse gas regulation, to flood mitigation and providing support for our sprawling cities. Therefore, protection of soil health from various pollutants including pesticides is important for its future sustenance. The pesticide Pymetrozine widely used in rice fields for the control of aphids and whiteflies is also registered in India for crop protection in Paddy. This study aimed to examine how long the pesticide Pymetrozine persists and how it breaks down in different types of soil in West Bengal. This research was conducted through laboratory experiments using LC-MS/MS at Bidhan Chandra Krishi Viswavidyalaya, Nadia. Applied two doses of Pymetrozine and analyzed soil samples over 30 days. Pymetrozine levels decreased by 95-97% after 15 days and fell below detectable levels after 30 days. Dissipation followed first-order kinetics, with a faster rate in new alluvial soil compared to red lateritic soil. This difference could be due to variations in soil properties, like organic carbon content and water retention.

Identification of the Csr global regulatory system mediated by small RNA decay in Aeromonas salmonicida.

We investigated the presence and functionality of the carbon storage regulator (Csr) system in Aeromonas salmonicida SWSY-1.411. CsrA, an RNA-binding protein, shared 89% amino acid sequence identity with Escherichia coli CsrA. CsrB/C sRNAs exhibited a typical stem-loop structure, with more GGA motifs, which bind CsrA, than E. coli. CsrD had limited sequence identity with E. coli CsrD; however, it contained the conserved GGDEF and EAL domains. Functional analysis in E. coli demonstrated that the Csr system of A. salmonicida influences glycogen biosynthesis, biofilm formation, motility, and stability of both CsrB and CsrC sRNAs. These findings suggest that in A. salmonicida, the Csr system affects phenotypes like its E. coli counterpart. In A. salmonicida, defects in csr homologs affected biofilm formation, motility, and chitinase production. However, glycogen accumulation and protease production were unaffected. The expression of flagellar-related genes and chitinase genes was suppressed in the csrA-deficient A. salmonicida. Northern blot analysis indicated the stabilization of CsrB and CsrC in the csrD-deficient A. salmonicida. Similar to that in E. coli, the Csr system in A. salmonicida comprises the RNA-binding protein CsrA, the sRNAs CsrB and CsrC, and the sRNA decay factor CsrD. This study underscores the conservation and functionality of the Csr system and raises questions about its regulatory targets and mechanisms in A. salmonicida.

Synergistic molecular mechanism of degradation in dye wastewater by Rhodopseudomonas palustris intimately coupled carbon nanotube - Silver modified titanium dioxide photocatalytic composite with sodium alginate

The intimately coupled photocatalysis and biodegradation (ICPB), which combined the advantages of high oxidation capacity of photocatalysis and high mineralization rate of biodegradation, has demonstrated excellent removal performance in the degradation of azo dyes with highly toxic, refractory, mutagenic and carcinogenic. In order to explore the metagenomics mechanism of the ICPB system, a novel ICPB was prepared by coupling Rhodopseudomonas palustris (R. Palustris), carbon nanotube - silver modified titanium dioxide photocatalytic composite (CNT-Ag -TiO2, CAT) and sodium alginate (SA) (R. palustris/CAT@SA, R-CAT). Metagenomics sequencing was used to investigate the molecular mechanism of adaptation and degradation of dyes by photosynthetic microorganisms and the adaptive and synergistic interaction between photosynthetic microorganisms and photocatalyst. Experiments on the adaptability and degradability of photosynthetic microorganisms have proved that low concentration azo dyes could be utilized as carbon sources for growth of photosynthetic microorganisms. Metagenomics sequencing revealed that R. palustris was the main degrading bacterium in photosynthetic microorganisms and the functional genes related to carbohydrate metabolism, biological regulation and catalytic activity were abundant. It was found that the addition of photocatalyst significantly up-regulated the functional genes related to the catabolic process, electron transport, oxidoreductase activity and superoxide metabolism of organic matter in the photosynthetic microorganisms. Moreover, many key gene such as alpha-amylase, 1-acyl-sn-glycerol-3-phosphate acyltransferase, aldehyde dehydrogenase enrichment in microbial basal metabolism, such as enoyl-CoA hydratase, malate dehydrogenase, glutathione S-transferase enrichment in degrading azo dyes and electron transport, and many key gene such as undecaprenyl-diphosphatase, carbon storage regulator, DNA ligase enrichment in response to dyes and photocatalysts were discovered. These findings would contribute to a comprehensive understanding of the mechanism of degradation of dye wastewater by ICPB system, a series of genes was produced to adapt to environmental changes, and played synergistic role in terms of intermediate product degradation and electron transfer for degrading azo dyes. The photosynthetic microorganisms might be a promising microorganism for constructing ICPB system.

Regulating ecosystem services in a local forest: Navigating supply, trade-offs, and synergies

The management of ecosystem service supply in local forests is closely linked to the ecological well-being of residents. This study aimed to estimate the supply, tradeoffs, and synergies of local forest-regulating ecosystem services using a land cover classification map (LCCM) and a forest type map (FTM). Rigorous literature reviews and expert Delphi analyses were conducted to elicit a list of key regulating services and evaluate the potential of each ecosystem type and category of the 1:5,000 LCCM and FTM to provide the selected services. The Getis-Ord Gi* analysis was combined with the land-sue scoring method to propose a method for estimating the supply, trade-offs, and synergies of local forest ecosystem services. The results revealed that the ranking order (1st to 5th) of the supply of regulating ecosystem services was as follows: erosion prevention, air quality regulation, heat island mitigation, water quality regulation, and carbon storage. When analyzing the correlation between the defined services of the entire city, almost all services showed a synergistic effect. However, when analyzed locally, spatial tradeoffs (between heat island mitigation and air quality regulation; carbon storage and air quality regulation; water quality regulation and air quality regulation) were observed in the eastern and northwestern forest areas. This suggests the need to consider not only the synergies and trade-offs between specific ecosystem services at the regional level but also at the local level when managing regulating ecosystem services of local forests. The results of this study can serve as important baseline references for planners and policymakers during the planning phase of policies aimed at managing the supply of regulating ecosystem services for forests with restricted access. Moreover, the study results can help refine the estimation of the supply of regulating ecosystem services by integrating additional data sources.

CsrA selectively modulates sRNA-mRNA regulator outcomes.

Post-transcriptional regulation, by small RNAs (sRNAs) as well as the global Carbon Storage Regulator A (CsrA) protein, play critical roles in bacterial metabolic control and stress responses. The CsrA protein affects selective sRNA-mRNA networks, in addition to regulating transcription factors and sigma factors, providing additional avenues of cross talk between other stress-response regulators. Here, we expand the known set of sRNA-CsrA interactions and study their regulatory effects. In vitro binding assays confirm novel CsrA interactions with ten sRNAs, many of which are previously recognized as key regulatory nodes. Of those 10 sRNA, we identify that McaS, FnrS, SgrS, MicL, and Spot42 interact directly with CsrA in vivo. We find that the presence of CsrA impacts the downstream regulation of mRNA targets of the respective sRNA. In vivo evidence supports enhanced CsrA McaS-csgD mRNA repression and showcases CsrA-dependent repression of the fucP mRNA via the Spot42 sRNA. We additionally identify SgrS and FnrS as potential new sRNA sponges of CsrA. Overall, our results further support the expanding impact of the Csr system on cellular physiology via CsrA impact on the regulatory roles of these sRNAs.

Diversified vegetation types on rangelands promote multiple soil‐based ecosystem services

AbstractRangelands have the potential to be provisioners of ecosystem services, including livestock products, carbon storage and greenhouse gas regulation, water and nutrient cycling, wildlife habitat, and biodiversity. Due to their vast extent and landscape heterogeneity, the degree to which different ecological components of rangelands contribute to ecosystem services can be varied. Soils are the foundation of rangeland health and associated ecosystem services. While many studies have examined the effect of grazing intensity on rangeland ecosystem services, few studies have looked at the broader rangeland landscape and how managing varying vegetation types can influence soil‐based ecosystem services. In this study, a suite of physical, chemical, and biological soil health indicators were measured in various vegetation types found within a working cattle ranch, including coastal live oak woodlands, coastal scrublands, annual grassland, and restored native perennial grassland. Based on the measured soil health indicators, results from this study show scrubland significantly diverges from other vegetation types, having higher water infiltration and plant available water, carbon stocks, and a more diverse microbial community that drives more dynamic cycling of carbon and nitrogen. Strategically maintaining scrubland on unproductive, highly erosive slopes downgradient of highly productive grassland areas could maintain forage production while protecting water quality and increasing carbon storage. These results highlight the relevance of holistically evaluating rangeland operations to assess soil function and ecosystem services and the potential risks and co‐benefits of varying vegetation types. Ultimately, process‐based linkages described here may provide a working example of how to manage ranches as functional mosaics of strategically maintained vegetation types.

CsrA coordinates the expression of ribosome hibernation and anti-σ factor proteins.

The Csr/Rsm system (carbon storage regulator or repressor of stationary phase metabolites) is a global post-transcriptional regulatory system that coordinates and responds to environmental cues and signals, facilitating the transition between active growth and stationary phase. Another key determinant of bacterial lifestyle decisions is the management of the cellular gene expression machinery. Here, we investigate the connection between these two processes in Escherichia coli. Disrupted regulation of the transcription and translation machinery impacts many cellular functions, including gene expression, growth, fitness, and stress resistance. Elucidating the role of the Csr system in controlling the activity of RNAP and ribosomes advances our understanding of mechanisms controlling bacterial growth. A more complete understanding of these processes could lead to the improvement of therapeutic strategies for recalcitrant infections.

Quorum sensing N-acyl homoserine lactones-SdiA enhances the biofilm formation of E. coli by regulating sRNA CsrB expression

As an important virulence phenotype of Escherichia coli, the regulation mechanism of biofilm by non-coding RNA and quorum sensing system has not been clarified. Here, by transcriptome sequencing and RT-PCR analysis, we found CsrB, a non-coding RNA of the carbon storage regulation system, was positively regulated by the LuxR protein SdiA. Furthermore, β-galactosidase reporter assays showed that SdiA enhanced promoter transcriptional activity of csrB. The consistent dynamic expression levels of SdiA and CsrB during Escherichia coli growth were also detected. Moreover, curli assays and biofilm assays showed sdiA deficiency in Escherichia coli SM10λπ or BW25113 led to a decreased formation of biofilm, and was significantly restored by over-expression of CsrB. Interestingly, the regulations of SdiA on CsrB in biofilm formation were enhanced by quorum sensing signal molecules AHLs. In conclusion, SdiA plays a crucial role in Escherichia coli biofilm formation by regulating the expression of non-coding RNA CsrB. Our study provides new insights into SdiA-non-coding RNA regulatory network involved in Escherichia coli biofilm formation.

Establishment of an In Bacterio Assay for the Assessment of Carbon Storage Regulator A (CsrA) Inhibitors.

Polymicrobial infections involving various combinations of microorganisms, such as Escherichia, Pseudomonas, or Yersinia, can lead to acute and chronic diseases in for example the gastrointestinal and respiratory tracts. Our aim is to modulate microbial communities by targeting the posttranscriptional regulator system called carbon storage regulator A (CsrA) (or also repressor of secondary metabolites (RsmA)). In previous studies, we identified easily accessible CsrA binding scaffolds and macrocyclic CsrA binding peptides through biophysical screening and phage display technology. However, due to the lack of an appropriate in bacterio assay to evaluate the cellular effects of these inhibitor hits, the focus of the present study is to establish an in bacterio assay capable of probing and quantifying the impact on CsrA-regulated cellular mechanisms. We have successfully developed an assay based on a luciferase reporter gene assay, which in combination with a qPCR expression gene assay, allows for the monitoring of expression levels of different downstream targets of CsrA. The chaperone protein CesT was used as a suitable positive control for the assay, and in time-dependent experiments, we observed a CesT-mediated increase in bioluminescence over time. By this means, the cellular on-target effects of non-bactericidal/non-bacteriostatic virulence modulating compounds targeting CsrA/RsmA can be evaluated.

Karst ecosystem services and land-covers dynamic: case study from Karst of Tuban and Tuban Regency, Jawa Timur, Indonesia

Karst area provides numerous ecosystem services from the interaction among its components of water, soil, and vegetation. However, a threat from unsustainable practice and resource overuses could diminish those services. Therefore, it is critical to steadily monitor the karst dynamics, especially in high-activity karst like the Karst of Tuban. This study aims to capture the karst dynamic, particularly the land-use change and how its impact on ecosystem services (ES). To achieve the research objective, an artificial intelligence tool of ARtificial Intelligence for Ecosystem Services (ARIES) is employed. ARIES examines a dataset for 6 years intervals beginning in 2012 to 2018 consisting of land-use and ecosystem change and ES using 150 m spatial resolution data. During that period, Tuban Regency undergoes a heavy change in artificial surface and a significant decrease in the water body and cropland. From the result, ARIES as the ES estimation tool can be used for robust calculation and preliminary study due to its limitation. The results also show the urge to address the issue of carbon storage and sediment regulation in the karst area and the land-cover changes in the Tuban Regency.

Spontaneous urban vegetation as an indicator of soil functionality and ecosystem services

AbstractQuestionsOur study focused on spontaneous vegetation in urban greenspaces in a Mediterranean city with the aim of relating plant community properties with ecological services along soil disturbance gradients. We asked which plant communities have the greatest plant biodiversity and soil carbon storage and the best‐performing nutrient cycles and water regulation.LocationMadrid City (Central Spain).MethodsWe studied four types of plant communities following soil disturbance gradients: vegetation on trampled soils, roadside vegetation, annual grasslands and perennial forbs. Regarding vegetation, we studied plant composition and productivity, plant diversity, plant growth forms and functional groups. Regarding soils, we determined soil organic carbon (TOC), available nutrients, the activity of seven enzymes relating to the main macronutrient cycles, and physical properties such as bulk density (BD) and soil water‐holding capacity (WHC). We used one‐way ANOVA to determine the influence of the plant community type on both soil and vegetation variables. Canonical correspondence analysis was performed to interpret the relationships between plant species assemblages with environmental gradients.ResultsPerennial forbs showed greater biomass and developed on soils with the greatest TOC and available phosphorus. Annual grasslands displayed the highest plant diversity. Roadside vegetation developed on soils with higher phenoloxidase activity when compared to vegetation on trampled soils and annual grasslands. Vegetation on trampled soils developed on soils with lower WHC, lower beta‐glucosidase, arylamidase and phosphatase activities and higher BD when compared to perennial forbs. Plant community distribution followed gradients most significantly associated with soil organic matter content, soil compaction and nutrient cycling performance.ConclusionsWe conclude that plant communities are good indicators of ecosystem function and services which are unevenly distributed throughout urban habitats. The management in Mediterranean unmaintained urban greenspaces should be aimed at avoiding soil compaction to promote biodiversity, carbon storage and water regulation.

E. coli Toxin YjjJ (HipH) Is a Ser/Thr Protein Kinase That Impacts Cell Division, Carbon Metabolism, and Ribosome Assembly.

Protein Ser/Thr kinases are posttranslational regulators of key molecular processes in bacteria, such as cell division and antibiotic tolerance. Here, we characterize the E. coli toxin YjjJ (HipH), a putative protein kinase annotated as a member of the family of HipA-like Ser/Thr kinases, which are involved in antibiotic tolerance. Using SILAC-based phosphoproteomics we provide experimental evidence that YjjJ is a Ser/Thr protein kinase and its primary protein substrates are the ribosomal protein RpmE (L31) and the carbon storage regulator CsrA. YjjJ activity impacts ribosome assembly, cell division, and central carbon metabolism but it does not increase antibiotic tolerance as does its homologue HipA. Intriguingly, overproduction of YjjJ and its kinase-deficient variant can activate HipA and other kinases, pointing to a cross talk between Ser/Thr kinases in E. coli. IMPORTANCE Adaptation to growth condition is the key for bacterial survival, and protein phosphorylation is one of the strategies adopted to transduce extracellular signal in physiological response. In a previous work, we identified YjjJ, a putative kinase, as target of the persistence-related HipA kinase. Here, we performed the characterization of this putative kinase, complementing phenotypical analysis with SILAC-based phosphoproteomics and proteomics. We provide the first experimental evidence that YjjJ is a Ser/Thr protein kinase, having as primary protein substrates the ribosomal protein RpmE (L31) and the carbon storage regulator CsrA. We show that overproduction of YjjJ has a major influence on bacterial physiology, impacting DNA segregation, cell division, glycogen production, and ribosome assembly.

Recognizing the Importance of an Urban Soil in an Open-Air City Museum: An Opportunity in the City of Madrid, Spain

This article examines the presence of urban soil buried under anthropogenic debris in an air-museum park in the Madrid city center (Spain), and highlights the particularities of this singular urbanized setting to indicate ecological evaluation options for soils. The study of a soil profile (with a thickness of about 2.30 m), classified as Urbic Technosols, allowed us to devise that it is formed by a series of filled-in amounts of artifacts (construction debris and other anthropogenic waste) of about 10–30%, plus organic and mineral materials. These soils’ composition and morphology depend on the natural conditions of the territory and also on anthropogenic activities. The soil properties (analyzed by conventional techniques) are moderate in acidity reaction and have relatively higher organic matter content. The Pb, Cu and Zn concentrations in anthropogenic horizons do not exceed the approximate permissible concentrations by 1.5–10-fold. Over the course of time, the soil profile has been transformed as a result of the impact of pedogenetic processes developing under the Mediterranean climate and man’s hand. Although urban environments present a certain complexity, at least the role of soil should be recognized regarding flood mitigation, recycling of wastes and toxins, filtering of nutrients or carbon storage and GHG regulation. The analysis of our results concludes the need to better perceive this soil profile and its green space to improve the urban ecosystem and to ensure better citizen well-being.

Long-Term Grazing Intensity Impacts Belowground Carbon Allocation and Mycorrhizas Revealed by 13CO2 Pulse Labeling

Despite the importance of grasslands for carbon storage and climate regulation, there is uncertainty about the effect of livestock grazing intensity on aboveground carbon assimilation and belowground carbon partitioning. Furthermore, the relationship between belowground carbon allocation and arbuscular mycorrhizal fungi, which serve as a conduit for carbon movement through the plant and soil, is unclear. To investigate this, we used an in situ 13C stable isotope pulse-chase labeling approach in plots under seven rates of sheep grazing intensity in a steppe grassland in northern China. We quantified the allocation of carbon to plants, soil, and soil-respired CO2 along with measurements of mycorrhizal hyphal density in the soil. With increasing grazing intensity, carbon assimilation per unit shoot biomass increased significantly, whereas carbon allocation to roots marginally decreased. Soil-respired CO2 appeared to be independent of grazing intensity. Mycorrhizal hyphal density decreased with increasing grazing intensity and was correlated significantly with new carbon input to roots 2 d after labeling and marginally related to that of soil 1 d after the 13C-CO2 pulse. Our study suggests that grazing intensity alters the distribution of carbon among different carbon pools within the plant-soil system. The results also underscored the key role of mycorrhizas as a fast route for carbon transfer from plant to soil.

Grasslands enhance ecosystem service multifunctionality above and below‐ground in agricultural landscapes

Abstract Managing agricultural landscapes integrate production, biodiversity conservation and the flow of ecosystem services (ES) is of paramount importance to simultaneously meet production goals and environmental challenges. However, the response of farmland biodiversity and multiple ES to land‐use change at multiple spatial scales remains poorly understood. We explored the effects of land‐use at local (grassland vs. oilseed rape fields) and landscape scale (cover of permanent grasslands) on the provision of biodiversity (plants, arthropods, birds), five ES (pollination, pest control, soil fertility, carbon storage and water regulation) and overall ES‐multifunctionality. ES‐multifunctionality was higher in grasslands than in crop fields, by 25.2% above‐ground and by 106.1% below‐ground. Multiple threshold analyses highlighted a particularly poor level of performance for below‐ground functions in crop fields. This habitat type was however capable of providing numerous above‐ground functions simultaneously, although at low levels of performance when compared to the maximum values recorded in the study. Grasslands supported higher biodiversity and provision of pollination, soil fertility, carbon storage and water regulation. Landscape composition influenced the provision of multiple ES: a 10% increase in grassland cover in the landscape enhanced above‐ground ES‐multifunctionality by 11.0% in both habitats. In particular, grasslands cover in the landscape supported the provision of arthropod diversity, pollination and pest control provided by carabids. Synthesis and applications. The results of this field study show the key importance of preserving semi‐natural grasslands in agricultural landscapes for the conservation of farmland biodiversity, for the protection of soils and the delivery of multiple ES critical for crop production. Maximization of multifunctionality necessitates the integration at the landscape scale (0.5–2 km) of semi‐natural patches within the intensively farmed agricultural matrix. This would require not only the protection of existing grasslands, but also their restoration in simplified landscapes. The promotion of mixed farming (i.e., both crop and livestock production) might increase semi‐natural grassland cover at the landscape scale.