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    Diagnosing delivery capabilities on a large international nature-based solutions project

    Nature-based solutions (NBS) are increasingly recognised as an effective response to a number of major urban challenges. These include heatwaves1,2,3, flooding4,5,6, water quality7,8 and public health and wellbeing9,10,11. While the concept of NBS emerged as recently as 201512, the idea of using urban nature to address these issues also features prominently in the more established fields of ecosystem services (which emerged in 2005)13 and green infrastructure (2002)14.Despite mounting evidence of their benefits, strategies built around NBS are seldom practically realised15; implementation in cities has been slow, inconsistent and often limited to demonstration sites16,17,18,19,20,21. For example, 6 years after Copenhagen embraced green infrastructure as a response to its acute flooding problems in 2011, the implementation of green infrastructure was only just ‘taking off’ in 2017, and remained highly contested22. Even retaining existing urban NBS remains a challenge; tree canopy cover, central to mitigation of urban heat island effects, is declining in many cities23,24. For example, metropolitan Melbourne experienced a loss of 2000 hectares between 2014 and 201825. In the US, an average of 36 million trees were lost each year from urban areas between 2009 and 201426.Barriers within the organisations responsible for implementing NBS are frequently identified as a primary reason for limited NBS delivery16,17,23,27,28,29. Delivery organisations have significant path dependencies; existing regimes are self-enforcing, and change is difficult30,31.The reasons for non-delivery have been characterised in detail in a broad range of literature. Barriers are highlighted in studies focused on urban forestry32,33, urban water management18,29,34, nature-based solutions35,36 and climate adaptation37,38. The issue has been investigated through the lenses of mainstreaming39,40, governance19,21,33,34,41,42, transitions31,36,43,44,45 and general analyses of barriers17,23,46,47. Papers describing the barriers to NBS have drawn on interviews with experts17,23,33,34,36,48, and direct project experiences49,50. At the time of writing, we are aware of nine review papers that present typologies of barriers to NBS delivery, based on systematic reviews of the considerable literature16,18,21,29,32,37,41,46,47.These studies have identified a largely consistent set of eight essential (and frequently lacking) traits for successful NBS implementation in local government. Leadership support is critical, both at the political and executive level18,20,21,29,34,41,51,52. A project team with the right capacity and timeframes to implement projects is also important32,37,51,53,54, as is a framework of internal mechanisms that facilitate the delivery of NBS, including clear approval processes, supportive policies and laws, and well-established standards for NBS design and maintenance16,18,20,35,47,55,56,57. A positive, supportive organisational culture for delivering new projects is also necessary, recognising that new NBS projects often have inherent (and novel) risks and trade-offs17,19,20,32,33,47,52,58. Finally, access to teams within the organisation that are both suitably skilled and supportive is vital16,18,23,46,54,59,60. Beyond the organisation itself, it is common for other levels of government to play an important role in approving aspects of NBS projects; an absence of support or clear process from higher regulatory authorities can pose a significant barrier32,33,38,44,51. Effective community engagement is also noted as important, recognising that many NBS need public support and/or private property owner consent to be successful23,37,38,46,49,61,62,63,64.While the barriers to NBS delivery have been the subject of significant attention, the implementation gap persists, with recent publications continuing to note the difficulty of NBS delivery33,36,52,57.A range of theoretical frameworks offer insight into how the implementation gap might be addressed. In the field of governance, The Policy Arrangement Model65 has been used to conceptualise governance in urban forestry32,33 and urban stormwater management21,34 as the temporary balance of a set of actors, discourses, rules and resources; changes to these variables may lead to changes in governance.In the Policy Arrangement Model, each of these four elements is significant, as is their interplay33. The actors included (or excluded) in a policy arrangement are crucial, given the range of agendas in typical stakeholders (e.g. politicians, community groups, chambers of commerce, financiers etc.), as are the relations between these actors (some may operate as coalitions, or as antagonists). Discourses include tacit and explicit conceptualisations of what the policy problem is, how it should be solved, and what values matter most. These are important in lending legitimacy to rules, which define interactions and roles between actors. These may be as formal as laws and design standards, or as informal as a set of undocumented organisational processes and norms (e.g. “talk to Anne in our compliance branch, she usually decides what is safe”). These elements are all vital in determining who deploys resources such as staff time, skills, budgets or equipment, and how they are deployed. Collectively, the dynamics between these four elements constitute a policy arrangement; changes in one element have the potential to affect others, and in turn spark shifts in governance65. However, these systems can be strongly entrenched66. Governance shifts are theorised to be typically driven by at least four factors: policy entrepreneurs (or ‘champions’), shock events, socio-political changes and ‘adjacent arrangements’ (developments in policy domains in related sectors or institutions)67.Policy entrepreneurs are also a focus of mainstreaming research, which highlights how these individuals advance NBS uptake by working within organisations to involve key stakeholders, engage citizens and contract technical expertise while incrementally introducing NBS considerations into planning practice35. This work is conceptualised as ‘horizontal’ mainstreaming, as officers champion NBS across their organisations, but it is argued that this must be supported by ‘vertical’ actions by top-down actors (such as executives and elected leaders) with the power to determine resource allocations and organisational structures39,40.To support NBS development and planning, the European Union’s Horizon 2020 programme initiated a series of large international demonstration projects, each involving collaborations between a number of cities, consultancies and universities. These include the UnaLab, ProGIreg, Connecting Nature, GrowGreen, Urban GreenUP and EdiCitNet projects68. These projects generally fund dedicated staff, as well as on-ground delivery of NBS, and have potential to address some or all of the barriers to NBS delivery that cities face. When considered in terms of the Policy Arrangement Model, the new actors, discourses and resources introduced by these projects all challenge the ‘temporary balance’ theorised to constitute the organisational status quo65. These projects also may encourage governance shifts67, by both facilitating the hiring of NBS policy entrepreneurs, and increasing a city’s exposure to influential adjacent arrangements in other centres of NBS expertise, such as university research units or exemplar municipalities. With the involvement of organisational champions/policy entrepreneurs, mainstreaming activities such as stakeholder outreach, citizen engagement and intra-organisational collaboration become increasingly possible35.This paper investigates the Horizon 2020 NBS project, Urban GreenUP. Urban GreenUP focuses on supporting partner cities to prepare NBS plans, as well as funding a multi-million Euro programme of investment in NBS interventions including floating vegetated islands, green walls on private structures, and streambank renaturalisation. The seven cities participating actively as project partners are Liverpool (UK), Ludwigsburg (Germany), Mantova (Italy), Valladolid (Spain), Izmir (Turkey), Quy Nhon (Vietnam) and Medellín (Colombia). This group of cities represents a wide range of governance arrangements and urban contexts in which NBS delivery occurs; Liverpool is a significant post-industrial centre emerging from sustained economic challenges compounded by government austerity, while Mantova has large areas of UNESCO world heritage and a legacy of industrial pollution. Quy Nhon is a coastal holiday town fairly new to NBS, while Ludwigsburg has extensive environmental legislation and has already successfully carried out major streambank restoration works on their local river. The former is largely governed by provincial government, with more operational management at a local level, whereas the latter has individual portfolio mayors, including one for the city’s environment. Valladolid has a population of 300,000 and a fairly compact urban form, whereas Medellin numbers over two million residents. We were able to work with each of these cities, effectively capturing the full range of capabilities and experiences in the Urban GreenUP project, and a significant variety of landscapes and organisational contexts in which NBS may be implemented.While the ‘generalisability’ of case studies is often limited, the constraints can be at least partially addressed through strategic sampling of cases69. Our sample is diverse, and while limited to seven cities, it does represent the full cohort of cities participating in this major EU programme designed to promote NBS innovation. A smaller sample size allows for a close, qualitative study of each case. Urban GreenUP presents a valuable opportunity to investigate the persistence of NBS barriers within local governments with ambitions for NBS implementation, with implications both for future innovation-oriented programmes such as Horizon 2020, and potentially the broader practice of NBS delivery in cities.Many cities beyond the Urban GreenUP group are preparing new NBS plans and programmes, and could benefit from insights arising from this study. Each GreenUP city is embarking on NBS planning and delivery and, at the time of our research, each had an individual or team employed with a specific NBS delivery role (with potential to serve the policy entrepreneur role highlighted in the literature). Local government often plays a key role in the implementation of urban NBS39,47,70, and Urban GreenUP places these organisations at its centre. Citizen engagement is an explicit focus of the project, as is the making of plans; these are both emphasised as opportunities for mainstreaming new practices35,71.We analyse the NBS implementation capacity of the cities within this study using an approach generally consistent with the practice of theory-based evaluation72,73. This ‘theory-based’ method breaks an implementation programme into its component elements, and assesses each element against available theory regarding what is required for success. This poses significant advantages over other evaluative methods because it focuses on the causative elements that lead to policy success or failure, rather than just the final outcomes achieved74, and is therefore more conducive to reforms of the institutional barriers discussed above.Theory-based evaluation typically takes place at the end of projects, but ours is ex ante; an approach noted by Weiss in her seminal outline of theory-based evaluation as having the potential to improve programme planning72. Evaluative practices have been noted as a particular weakness in local government NBS programmes, both at the political and officer level, due to a fear that acknowledging problems would lead to criticism of failures36. We sought to mitigate this issue both through use of an ex ante approach, and by designing a tool that creates distance between evaluators and individual practitioners.This paper investigates the enduring difficulties faced by cities seeking to deploy NBS, in the context of a major NBS project spanning seven cities. We had two key research questions. First, do case study cities have the capabilities required for successful NBS delivery, or are there barriers that continue to make this difficult? Second, does identifying and measuring a city’s NBS delivery capabilities facilitate improvements in these capabilities?We elicited organisational barriers from NBS practitioners in participating cities using a purpose-built diagnostic tool. We used this approach because tools that enable organisations to learn about their success factors have the potential to address implementation gaps44,71.The tool was developed to bring lessons from the literature into an operational context, by enabling practitioners to assess and rate their organisation’s capability levels across eight key areas, such as political support, alignment between teams, and technical knowledge (refer to Table 1). The tool posed a series of questions pertaining to each of these eight capability areas; users answered by selecting from a set of pre-defined answers. The tool associated each answer with a level of capability, which was reported as a final assessment of an organisation’s capabilities. This was provided to the practitioner directly on completion of the tool’s questions, enabling an immediate estimate of their organisation’s NBS delivery capacity, and a diagnosis of any key barriers they would be likely to face in future NBS projects. These results formed the basis for our reflections on NBS delivery capacity within Urban GreenUP, as well as discussions with practitioners to understand how results of the tool were received.Table 1 Eight success factors for urban NBS in local government.Full size tableOur research proceeded in three steps. First, we drew on the literature to define a set of eight key capabilities—which we call ‘success factors’—for NBS; these formed the basis of the tool. Second, practitioners within NBS teams in participating cities used the tool to identify their capability levels. Finally, we interviewed users to evaluate the impact of the tool. More

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    Holocene life and microbiome profiling in ancient tropical Lake Chalco, Mexico

    Evaluation of the lithological, geochemical, and fossil diatom evidenceWe obtained a short, 270-cm sediment core ( 250 m40. Lake Chalco lies in the central region of the Trans-Mexican Volcanic Belt (Fig. 1). It is bounded to the north by the Sierra de Santa Catarina, to the east by the Sierra Nevada (Volcanoes Popocatepetl, Iztaccíhuatl, Tláloc and Telapón), and to the west, by Volcano Teuhtli, which is the closest volcano to the waterbody (~ 6.5 km)41. Lake sediments are composed mainly of clays that have been described as impermeable. The Lake Chalco Basin, however, is a highly complex system and seismically active. Therefore, the presence of active fractures during the Holocene is possible. Active fractures may result in inflow from the deep aquifer. Historical data indicate the existence of freshwater springs in upper parts of the aquifer, mainly at the eastern piedmont42. For instance, mineral and thermal waters at Peñón del los Baños, near the Mexico City airport (33 km from Lake Chalco), have been reported since Aztec times around AD 1,325. This spring is associated with a system of seismically active fractures. No thermal waters have been reported in modern Lake Chalco, however phreatomagmatic activity ( > 100,000 years BP) from the Xico Volcano has been documented43. Recent studies showed that the water table position changes from the upper parts of the watershed to central Texcoco, 45 km from Lake Chalco. In this study four components of the flow system were identified, including waters of recent infiltration and local circulation, evidencing intermediate chemical evolution, and waters more chemically evolved of large flow trajectories and of deep circulation44.The lithology of the studied sediment sequence is as follows: (1) Sediments from 270 to 250 cm are characterized by lapilli, ash and black to light brown silty ashes (massive to stratified); (2) Sediments in the interval 250–235 cm are composed of diatomite (yellow); (3) From 235 to 200 cm, sediments are characterized by massive black to brown sandy silts (brown); (4) From 200 to 70 cm, sandy brown to reddish, banded laminated silts, with scattered or banded pumice fragments (red) are present; (5) Sediments from 70 to 60 cm are black silty sands with organic material; (6) From 60 to 50 cm sediments are sandy brown to reddish, laminated silt, with scattered or banded pumice fragments (red); (7) Sediments from 50 to 40 cm are characterized by lapilli, ash and black to light brown silty ashes, massive to stratified, and (8) uppermost sediments from 40 to 0 cm are black silty sands with organic material (Fig. 3)36.Figure 3Taxonomic diversity revealed by metagenomic and fossil diatom analysis, and geochemical variables from the Lake Chalco Holocene sediment sequence. Each horizontal bar represents a collected sample, with the exception of the upper row, which shows the average of surface samples S1 (0 cm, i.e., modern) and S2 (0 cm, i.e., modern) (Supplementary Table S1). The lithology of the 270-cm sediment sequence is shown in the first column. The Upper Toluca Pumice (UTP) is represented as tephra underneath the first column (from left to right). Taxonomic diversity is depicted as the relative abundance of phyla Bacteria (green), Archaea (pink), and Eukarya (blue) (columns 2–4). Percent values correspond to the diversity of peptide sequences corresponding to each domain. Geochemical variables related to biological processes and past conditions are shown in columns 5–7. Results of fossil diatom analysis are shown in column 8. Dark horizontal lines show the boundaries for each delimited paleoenvironmental zone: (1) freshwater, (2) hyposaline and (3) subsaline. Edited in CorelDRAW 2020 version 22.0.Full size imageElemental geochemistry and fossil diatoms in sediment cores can be used as indicators of past wet and dry climate intervals. We measured geochemical indicators including element concentrations and ratios, and Total Organic Carbon (TOC) throughout the core (Supplementary Table S1). Total organic carbon is an important component of sediments and soils and can be used to assess the environmental status of terrestrial and aquatic ecosystems45. Maximum TOC values characterize the period of hyposaline conditions. The Mn/Fe ratio, often used to track past O2 content in bottom waters and changing redox conditions, was used as a proxy for water-column oxygen concentration46. The Mn/Fe ratios display highest values at depths of 200, 150 and 60 cm, suggesting periods of permanent anoxia during warmer conditions and excessive nutrient inputs47. The Fe/Ti ratio provides information about fluvial sediment sources. Changes in the abundance of iron oxides can be used to infer fluctuations in inputs of land‐derived detrital material48. We observed increasing Fe/Ti ratio values in superficial layers, and highest values at 100 cm. We performed cluster analysis based on Euclidean distance which revealed three groups of samples (Supplementary Fig. S1).We identified three zones in the Lake Chalco Holocene sediment sequence, based on geochemical analysis and diatom assemblages, which reflect different paleoenvironmental conditions: (1) a cool, freshwater lake (235–210 cm), (2) a warm, hyposaline lake (185–60 cm), and (3) a temperate, subsaline lake (50–0 cm) (Figs. 3, 5, Supplementary Fig. S1, Table S1).Fossil diatom assemblages provide information about past environmental changes and water quality7,49. Fossil diatom analysis, along with knowledge of species ecological preferences, enables inference of past limnological variables such as temperature, salinity, pH, electrical conductivity, and phosphorus concentration7. Inferences from our fossil diatom record concur with an earlier diatom-based paleoclimate study from Lake Chalco. Our studies revealed that during the last deglacial (~ 19,500–11,500 cal years BP), conditions were colder and much wetter than present. Assemblages are dominated by small araphid diatoms Gomphonema affine and Cocconeis placentula (Fig. 3). From 11,500 to 4,500 cal years BP, Lake Chalco was characterized by hyposaline conditions, with higher evaporation rates until ~ 6,500 cal years BP. Typical diatoms taxa include Anomoeoneis costata, Halamphora veneta and small araphids. After ~ 6,500 cal years BP, salinity in Lake Chalco declined, mean annual precipitation increased slightly, and summer insolation, seasonality, and evaporation decreased7. Assemblages are composed of H. veneta, Nitzschia frustulum, Cyclotella meneghiniana and small araphid diatoms.Meta-taxonomic analysis of Prokaryote and Eukaryote diversityOur metagenomic analysis identified 36,722 OTUs (genera) in the sediments of Lake Chalco. Among those genera, 81% correspond to bacteria (29,818 ± 106 identified to genera [ig]), 15% to Archaea (5,710 ± 118 ig), 3% to Eukarya (1,147 ± 6 ig), and 76. Iron is considered a potentially harmful element (PHE), which may be indicative of human-mediated contamination. For instance, high Fe concentrations in surficial sediments could be related to inputs of clastic sediments, and often reflect agricultural activities77. We determined 13 plant genera, five belonging to the family Poaceae (58%), including the genus Zea (corn), known to have been cultivated and consumed by early settlers78, and Oryza, a fast-growing weed that is indicative of human-mediated habitat disturbance. Twelve microscopic fungal genera were observed, including taxa that are pathogenic on wheat and rice (Gibberella and Cladochytrium), plants, keratin, and flies (Supplementary Fig. S10)79, and a protozoan that is pathogenic in humans (Giardia). We found high abundances of the family Culicidae (18%) (Supplementary Fig. S10) during the period of human occupation. The subsaline zone displays 290 unique pfams and the highest number of representatives of potassium metabolism throughout the entire Holocene (Supplementary Fig. S4B). The abundance of genes related to Cyanobacteria in this zone is much lower (9%), in contrast with findings from the Blastp against MetaProt database of the freshwater (30%) and hyposaline zones (60%) (Supplementary Fig. S10, Table S3).Our findings suggest that the biota in and around Lake Chalco during the Holocene responded mainly to changes in temperature, salinity, and trophic state, reflecting climate and human impacts over the last 6,000 years. This implies landscape modifications, agricultural activities and accelerated lake eutrophication. Furthermore, prokaryotic assemblages revealed gradual deposition of microbial communities capable of anaerobic fermentation of organic material and methanogenesis, as well as evidence for volcanic activity, inferred from the metabolic potential for sulfur cycling in the deeper zones (hyposaline and freshwater). This study generated information on Neotropical Prokaryote and Eukaryote diversity and microbial metabolic pathways during the Holocene. Nevertheless, we recommend that future studies focus on detailed characterization of microbial substrates and constrain post-depositional processes. Such studies should also consider selection processes that result from gradual depletion of substrates during burial52,59,63. Finally, we highlight the importance of including additional geochemical measures, such as porewater chemistry80. and sedimentology81 in future studies and measuring biomass by qPCR assay. More

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    Particulate organic matter as a functional soil component for persistent soil organic carbon

    Study site and soil samplingThe soil was collected at 5 − 20 cm (Ap horizon) from an agricultural field in Southern Germany (Freising, Bavaria, 48°23’53.8“N, 11°38’39.7“E) in December 2017. The sampling area is situated within the lower Bavarian upland, and characterized by a mean annual temperature of 7.8 °C and mean annual precipitation of 786 mm. The soil type is a Cambisol (silty clay loam; 32% clay, 53% silt, and 14% sand) with a considerable amount of loess mixed with underlying Neogene sandy sediments. The soil was selected to represent a widely distributed soil type and land use. The collected soil was oven-dried (2 days, 40 °C), sieved ( More

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    Microbially facilitated nitrogen cycling in tropical corals

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    Evaluation on soil fertility quality under biochar combined with nitrogen reduction

    Research areaThe study was conducted in the Yunyang Experimental Station (108° 54′ E, 30° 55′ N; altitude of 700 m), Southwest University, Chongqing, China. The study area has a subtropical monsoon humid climate with an average annual sunshine duration of 1500 h, average annual temperature of 18.4 °C average annual rainfall of 1100.1 mm, and the rain period predominantly prolongs from June to September. Local soil type is clay loam in texture and Dystric Purple-Udic Cambosols according to the Chinese Soil Taxonomy (CRGCST 2001). Basic properties of 0–20 cm soil layer were as follows: pH 7.29, total N 0.94 g kg−1, total C 7.14 g kg−1, available N 37.45 mg kg−1, available P 2.36 mg kg−1, and available K 72.58 mg kg−1, respectively.The tested biochar was purchased from the Nanjing Qinfeng Straw Technology Co., Ltd. (Nanjing, China), which was made by pyrolysis of the rice (Oryza sativa L.) straw with limited oxygen supply at 500 °C for 2 h. Its properties were as follows: total N 0.61 g kg−1, total P 1.99 g kg−1, total K 27.15 g kg−1, total C 537.97 g kg−1 and pH 8.70.Experimental designA two-year filed experiment (2017–2019) was performed in a completely randomized design with twelve treatments in triplicates including two factors. The first factor was the application of biochar including B0 (0 t ha−1), B10 (10 t ha−1), B20 (20 t ha−1) and B40 (40 t ha−1); and the second factor is the application level N fertilizer including conventional rate (application amount by local farmers)-180 kg N ha−1 (N100), 80% of conventional rate-144 kg N ha−1 (N80) and 60% of conventional rate-108 kg N ha−1 (N60). The plot size was 3 m × 6 m with a border (0.5 m wide) between plots. Biochar was applied to soil only in the first year before the sowing of rapeseed. Each treatment plot received the same amount of potassium (90 kg K2O ha−1) and phosphorus (90 kg P2O5 ha−1). Further details of fertilizer application have been reported by Tian et al.24, being the same for the two-year experiment. Weed, pesticide, and pest management kept the same with the local farmers’ rapeseed management practices. Winter rapeseed (Sanxiayou No.5) was used in the experiment, which was sowed on 21 October 2017 and on 16 October 2018, respectively, and was harvested on 1 May in both years (2018 and 2019).Sampling and analysis of soil and cropCrop yieldRapeseed was hand-harvested when 70–80% of total seeds changed their color from green to black on 1 May 2019, and each plot was separately harvested for seed yield. Seed yield was calculated using 6% as standard seed moisture content.Soil indicesAfter the rapeseed harvest, soil samples were collected from all plots. Five sampling points were randomly selected within each plot. At each point, twenty soil cores of 2.5 cm diameter and 20.0 cm depth were taken in a 1 m radius of the point. All soil cores from each point were put in a plastic bag and thoroughly bulked, crumbled and mixed for physical, chemical and biological analyses. By dividing each soil sample into two subsamples, one subsample was ground, passed through a 2-mm sieve and was air-dried for the analyses of soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP), total potassium (TK), alkali-hydrolyzale nitrogen (AN), available phosphorus (AP), available potassium (AK)25, particulate organic carbon (POC), water-soluble organic carbon (DOC), easily oxidized organic carbon (AOC)26, sucrase (SUC) and urease (URE)27, and another one was ground, passed through a 2-mm sieve and was stored in a refrigerator at − 20 °C for the analyses of structural and functional characteristics of soil microbial community28. At the same time, mixed soil samples (0–20 cm) from five points in each plot were taken using a shovel for soil aggregates analyses24.Drying method was used to determine soil water content (SWC); soil temperature (ST) was measured by temperature probe on the LI6400–09 (LI-COR Inc., Lincoln, NE); potassium dichromate oxidation method was used to determine SOM and DOC content; TN was measured by the Kjeldahl method; TP was determined by Mo-Sb colorimetric method; TK was determined by NaOH melting and analyzed using an atomic spectrophotometry; AN was determined by diffusion-absorption method; AP was quantified by colorimetric analysis following extraction of soil with 0.5 mol L−1 NaHCO3; AK was measured using 1.0 mol L−1 CH3COONH4 extraction; POC was determined by sodium hexametaphosphate dispersion method; AOC was measured by potassium permanganate oxidation method; SUC was measured by 3,5-dinitrosalicylic acid colorimetric determination method; URE was measured by phenol-sodium hypochlorite indophenol colorimetry method; amount of bacteria (B), fungi (F), actinomycetes (A), gram-positive bacteria (GP), gram-negative bacteria (GN) was measured by the Bligh–Dyer method; utilization of sugars (S), amino acids (AA), phenolic acids (PA), carboxylic acids (CA), amines (AM) and polymers (P) by microorganism was measured using commercial Biolog EcoPlate (Biolog Inc., CA, USA).Shannon index (H), Simpson index (D), and evenness index (E) were calculated by the following equations:$$ {text{AWCD}} = sum {(C_{i} – R_{i} )} /n $$$$ {text{H}} = – sum {P_{i} } (ln P_{i} )quad P_{i} = (C_{i} – R_{i} )/sum {(C_{i} – R_{i} } ) $$$$ {text{D}} = 1 – sum P _{i}^{2} $$$$ {text{E}} = {text{H}}/ln {text{S}} $$where n is the 31 carbon sources on the ECO board; Ci and Ri and are the optical density values of the microwell and the control well respectively; Pi is the ratio of the absorbance of a particular well i to the sums of absorbance of all 31well at 120 h; S is the number of color change holes, which represents the number of carbon source used by the microbial community; Average well color development (AWCD), representing the overall carbon substrate utilization potential of cultural microbial communities across all wells per plate.In order to investigate the aggregate structure, all bulk clod samples from each plot were carefully mixed and then gently sieved to pass through a 10-mm sieve. According to the wet-sieving and dry-sieving protocol, the tested soil was fractionated into  > 5, 2 ~ 5, 1 ~ 2, 0.25 ~ 1 and  0.25} right)} }}{{sumnolimits_{{i = 1}}^{n} {(w_{i} )} }} times 100% $$$$ {text{D – MWD}}left( {{text{W – MWD}}} right) = sumlimits_{{i = 1}}^{n} {(bar{d}_{i} w_{i} )} $$$$ {text{D – GMD}}left( {{text{W – GMD}}} right) = exp left[ {frac{{sumlimits_{{i = 1}}^{n} {m_{i} ln bar{d}_{i} } }}{{sumlimits_{{i = 1}}^{n} {m_{i} } }}} right] $$where DR0.25 and WR0.25 are the proportion of  > 0.25 mm soil mechanical-stable aggregates and water-stable aggregates, respectively; D-MWD and W-MWD are the mean weight diameter of mechanical-stable aggregates and water-stable aggregates (mm), respectively; D-GMD and W-GMD are the mean geometric diameter of mechanical-stable aggregates and water-stable aggregates (mm), respectively; mi is mass in size fraction i; and wi is the proportion (%) of the total sample mass in size fraction i and di is mean diameter of size fraction i.Evaluation of soil fertilityGrey correlation analysisGrey correlation analysis refers to a method of quantitative description and comparison of a system’s development and change. The basic idea is to determine whether they are closely connected by determining the geometric similarity of the reference data column and several comparison data columns, which reflects the degree of correlation between the curves29. The grey relational coefficient ξi (k) can be expressed as follows:$$ xi (k) = frac{{mathop {min }limits_{i} mathop {min }limits_{k} left| {x_{0} (k) – x_{i} (k)} right| + rho mathop {max }limits_{i} mathop {max }limits_{k} left| {x_{0} (k) – x_{i} (k)} right|}}{{left| {x_{0} (k) – x_{i} (k)} right| + rho mathop {max }limits_{i} max left| {mathop {x_{0} (k)}limits_{k} – x_{i} (k)} right|}} $$$$ x_{i}^{k} = frac{{x_{i}^{k} }}{{mathop {max }limits_{i} x_{i}^{k} }} $$$$ gamma _{i} = frac{1}{n}sumlimits_{{k = i}}^{n} {xi _{i} } (k) $$$$ omega _{{i(gamma )}} = frac{1}{n}sumlimits_{{i = 1}}^{n} {gamma _{i} } $$$$ G_{i}^{k} = sumlimits_{{i = 1}}^{n} {left( {xi _{i} times omega _{{i(gamma )}} } right),quad k = 1,2,3, ldots ,n;quad i = 1,2,3, ldots ,n} $$where (x_{i}^{k}) The i trait observation value of treatment k; (mathop {max }limits_{i} x_{i}^{k}) The maximum value of the i trait in all treatments; (mathop {min }limits_{i} x_{i}^{k}) The minimum value of the i trait in all treatments; (mathop {min }limits_{i} mathop {min }limits_{k} left| {x_{0} (k) – x_{i} (k)} right|) Second level minimum difference; (mathop {max }limits_{i} mathop {max }limits_{k} left| {x_{0} (k) – x_{i} (k)} right|) Second level maximum difference; (rho) Resolution coefficient (0.5).Principal component analysisPrincipal component analysis refers to a multivariate statistical method that converts multiple indicators into several comprehensive indicators by the idea of dimensionality under the premise of losing little information. It simplifies the complexity in high-dimensional data while retaining trends and patterns30.Cluster analysisCluster analysis comprises a range of methods for classifying multivariate data into subgroups. Using the euclidean distance as a measure of the difference in the fertility of each treatment, the shortest distance method was used to systematically cluster according to the degree of intimacy and similarity of soil fertility levels. By organizing multivariate data into such subgroups, clustering can help reveal the characteristics of any structure or patterns present31.Statistical analysisCorrelation analysis was performed to assess the relationships between rapeseed yield and soil attributes. Grey correlation analysis and principal component analysis were performed to establish comprehensive score for soil fertility and the main soil factors affecting rapeseed yield. Cluster analysis was used to cluster the soil fertility of each treatment. All the statistical analyses were performed using Excel 2018 (Office Software, Inc., Beijing, China) and SPSS 17.0 (SPSS Inc., Chicago, Illinois, USA). The comparisons of treatment means were based on LSD test at the P  More