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This method employs liquid chromatography-tandem mass spectrometry to rapidly quantify chitin-derived glucosamine for estimating fungal biomass. Analyte retention was achieved using hydrophilic interaction liquid chromatography, with a zwitter-ionic stationary phase (ZIC-HILIC), and isocratic elution using 60% 5 mM ammonium formate buffer (pH 3.0) and 40% ACN. Inclusion of muramic acid and its chromatographic separation from glucosamine enabled calculation of the bacterial contribution to the latter. Galactosamine, an isobaric isomer to glucosamine, found in significant amounts in soil samples, was also investigated. The two isomers form the same precursor and product ions and could not be chromatographically separated using this rapid method. Instead, glucosamine and galactosamine were distinguished mathematically, using the linear relationships describing the differences in product ion intensities for the two analytes. The m/z transitions of 180 → 72 and 180 → 84 were applied for the detection of glucosamine and galactosamine and that of 252 → 126 for muramic acid. Limits of detection were in the nanomolar range for all included analytes. The total analysis time was 6 min, providing a high sample throughput method. [ABSTRACT FROM AUTHOR]

A liquid chromatography with electrospray ionization mass spectrometry method was developed to quantitatively and qualitatively analyze 13 hydroxamate siderophores (ferrichrome, ferrirubin, ferrirhodin, ferrichrysin, ferricrocin, ferrioxamine B, D1, E and G, neocoprogen I and II, coprogen and triacetylfusarinine C). Samples were preconcentrated on-line by a switch-valve setup prior to analyte separation on a Kinetex C18 column. Gradient elution was performed using a mixture of an ammonium formate buffer and acetonitrile. Total analysis time including column conditioning was 20.5 min. Analytes were fragmented by applying collision-induced dissociation, enabling structural identification by tandem mass spectrometry. Limit of detection values for the selected ion monitoring method ranged from 71 pM to 1.5 nM with corresponding values of two to nine times higher for the multiple reaction monitoring method. The liquid chromatography with mass spectrometry method resulted in a robust and sensitive quantification of hydroxamate siderophores as indicated by retention time stability, linearity, sensitivity, precision and recovery. The analytical error of the methods, assessed through random-order, duplicate analysis of soil samples extracted with a mixture of 10 mM phosphate buffer and methanol, appears negligible in relation to between-sample variations. [ABSTRACT FROM AUTHOR]

In this work the distribution of free and weekly adsorbed aromatic acids (phthalic acid and ten phenolic acids; gallic, p-hydroxybenzoic, salicylic, vanillic, protocatechuic, p-coumaric, syringic, sinapic, ferulic and caffeic acid), which could participate in weathering and soil formation processes, were studied for O, E and B horizons in a podzol soil in central Sweden. For the analysis a simple and rapid quantitative and qualitative liquid chromatography-tandem mass spectrometry method (using gradient elution) was developed with LODs ranging from 5 to 25 nM. Different soil solution sampling techniques (tension-lysimeter and soil centrifugation) and soil extraction with either 10 mM phosphate buffer (pH7.2) or 50:50 (v/v) 10 mM phosphate buffer:MeOH were compared. All eleven acids were found in detectable or trace concentrations. The most abundant aromatic acids were vanillic and phthalic acid with concentrations around 1 μM for O and E horizon respectively. Lysimeter samples resulted in the lowest concentrations followed by centrifugation samples. In general, buffer:MeOH extraction resulted in the highest concentrations for the O horizon, likely due to MeOH's ability to compete for hydrophobic sites on soil organic matter (SOM). Then again, pure buffer with its higher ion strength, interfering with the acids electrostatic interactions with clay particles, leads to higher extracted concentrations for the E and B horizons. Since the efficiency of the extraction solutions, to a large extent, depends on the sample properties, a general approach is hard to appoint. However, the extraction of substituted cinnamic acids is in general facilitated by adding MeOH to the extraction solution. The use of statistical methods for the evaluation of the results showed a large and significant difference in aromatic acid concentrations received using different sampling techniques and sample preparations. In fact, sampling methods resulted in higher variations in aromatic acid concentrations than sampled horizon. [ABSTRACT FROM AUTHOR]

Societal Impact Statement: Efficient mitigation of climate change requires predictive models of forest ecosystems as sinks for atmospheric carbon. Mycorrhizal fungi are drivers of soil carbon storage in boreal forests, yet they are typically excluded from ecosystem models, because of a lack of information about their growth and turnover. Closing this knowledge gap could help us better predict future responses to climate change and guide policy decisions for sustainable management of forest ecosystems. This study provides new estimates of the production and turnover of mycorrhizal mycelial biomass and necromass. This information can facilitate the integration of mycorrhizal fungi into new predictive models of boreal forest soils. Summary: In boreal forests, turnover of biomass and necromass of ectomycorrhizal extraradical mycelia (ERM) are important for mediating long‐term carbon storage. However, ectomycorrhizal fungi are usually not considered in ecosystem models, because data for parameterization of ERM dynamics is lacking.Here, we estimated the production and turnover of ERM biomass and necromass across a hemiboreal Pinus sylvestris chronosequence aged 12 to 100 years. Biomass and necromass were quantified in sequentially harvested in‐growth bags, and incubated in the soil for 1–24 month, and Bayesian calibration of mathematical models was applied to arrive at parametric estimates of ERM production and turnover rates of biomass and necromass.Steady states were predicted to be nearly reached after 160 and 390 growing season days, respectively, for biomass and necromass. The related turnover rates varied with 95% credible intervals of 1.7–6.5 and 0.3–2.5 times yr−1, with mode values of 2.9 and 0.9 times yr−1, corresponding to mean residence times of 62 and 205 growing season days.Our results highlight that turnover of necromass is one‐third of biomass. This together with the variability in the estimates can be used to parameterize ecosystem models, to explicitly include ERM dynamics and its impact on mycorrhizal‐derived soil carbon accumulation in boreal forests. [ABSTRACT FROM AUTHOR]