Soil and plant inoculation with heterotrophic zinc-solubilizing bacteria (ZSB) is considered a promising approach for increasing zinc (Zn) phytoavailability and enhancing crop growth and nutritional quality. Nevertheless, it is necessary to understand the underlying bacterial solubilization processes to predict their repeatability in inoculation strategies. Acidification via gluconic acid production remains the most reported process. In this study, wheat rhizosphere soil serial dilutions were plated on several solid microbiological media supplemented with scarcely soluble Zn oxide (ZnO), and 115 putative Zn-solubilizing isolates were directly detected based on the formation of solubilization halos around the colonies. Eight strains were selected based on their Zn solubilization efficiency and siderophore production capacity. These included one strain of , two of, three strains of , one of, and one strain of In ZnO liquid solubilization assays, the presence of glucose clearly stimulated organic acid production, leading to medium acidification and ZnO solubilization. While solubilization by and was attributed to the accumulated production of six and seven different organic acids, respectively, the other strains solubilized Zn via gluconic, malonic, and oxalic acids exclusively. In contrast, in the absence of glucose, ZnO dissolution resulted from proton extrusion (e.g., via ammonia consumption by strains) and complexation processes (i.e., complexation with glutamic acid in cultures of ). Therefore, while gluconic acid production was described as a major Zn solubilization mechanism in the literature, this study goes beyond and shows that solubilization mechanisms vary among ZSB and are strongly affected by growth conditions. Barriers toward a better understanding of the mechanisms underlying zinc (Zn) solubilization by bacteria include the lack of methodological tools for isolation, discrimination, and identification of such organisms. Our study proposes a direct bacterial isolation procedure, which prevents the need to screen numerous bacterial candidates (for which the ability to solubilize Zn is unknown) for recovering Zn-solubilizing bacteria (ZSB). Moreover, we confirm the potential of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) as a quick and accurate tool for the identification and discrimination of environmental bacterial isolates. This work also describes various Zn solubilization processes used by wheat rhizosphere bacteria, including proton extrusion and the production of different organic acids among bacterial strains. These processes were also clearly affected by growth conditions (i.e., solid versus liquid cultures and the presence and absence of glucose). Although highlighted mechanisms may have significant effects at the soil-plant interface, these should only be transposed cautiously to real ecological situations.

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Interactions between model parameters and low spatiotemporal resolution of available data mean that conventional soil organic carbon (SOC) models are often affected by equifinality, with consequent uncertainty in SOC forecasts. Estimation of belowground C inputs is another major source of uncertainty in SOC modelling. Models are usually calibrated on SOC stocks and fluxes from long‐term experiments (LTEs), whereas other point data are not used for constraining the model parameters. We used data from an agricultural long‐term (>65 years) fertilization experiment to test a multi‐objective parameter estimation approach on the RothC model, combining SOC data from different fertilization treatments with microbial biomass, basal respiration and Zimmermann's fractions data. We also compared two methods to estimate the belowground C inputs: a conventional scaling of belowground biomass from crop harvest yield and an alternative approach based on constant belowground C for cereals measured experimentally in the field. The resulting posterior parameter distributions still suffered from some equifinality; the most stable C pool kinetic constants and composition of exogenous organic matter were the most sensitive parameters. The use of fixed belowground C inputs for cereals improved the model performance, reducing the importance of treatment‐specific parameters and processes. The introduction of microbial biomass and basal respiration data was effective for increasing determination of the calibration, but also suggested a change in the model structure: the microbial biomass pool, which is proportional to the C inputs in the traditional models, could be represented by different microbial physiology functions. Highlights Multi‐objective calibration with SOC and microbial or soil fractionation data can reduce model uncertainty. We compared different methods to estimate belowground C inputs on a long‐term trial in Switzerland. Fixed belowground C inputs measured in the field gave the best model performance. Microbial data can improve model calibration, but a change in model structure is suggested.

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Purpose Green manuring can increase the plant available fraction of zinc (Zn) in soil, making it a potential approach to increase wheat Zn concentrations and fight human Zn deficiency. We tested whether green manure increases the ability of both the native soil bacteria and inoculated Zn solubilizing bacteria (ZSB) to mobilize Zn. Methods Wheat was grown in a pot experiment with the following three factors (with or without); (i) clover addition; (ii) soil x-ray irradiation (i.e. elimination of the whole soil biota followed by re-inoculation with the native soil bacteria); and (iii) ZSB inoculation. The incorporation of clover in both the irradiated and the ZSB treatments allowed us to test green manure effects on the mobilization of Zn by indigenous soil bacteria as well as by inoculated strains. Results Inoculation with ZSB did neither increase soil Zn availability nor wheat Zn uptake. The highest soil Zn availabilities were found when clover was incorporated, particularly in the irradiated soils (containing only soil bacteria). This was partly associated with the stimulation of bacterial activity during the decomposition of the incorporated green manure. Conclusion The results support that the activity of soil bacteria is intimately involved in the mobilization of Zn following the incorporation of green manure.