『Abstract
Amino sugars in fungal cell walls (such as chitin) represent
an important source of nitrogen (N) in many forest soil ecosystems.
Despite the importance of this material in soil nitrogen cycling,
comparatively little is known about abiotic and biotic controls
on and the timescale of its turnover. Part of the reason for this
lack of information is the inaccessibility of these materials
to classic bulk extraction methods. To address this issue, we
used advanced visualization tools to examine transformation pathways
of chitin-rich fungal cell wall residues as they interact with
microorganisms, soil organic matter and mineral surfaces. Our
goal was to document initial micro-scale dynamics of the incorporation
of 13C- and 15N-labeled chitin into fungi-dominated
microenvironments in O-horizons of old-growth forest soils. At
the end of a 3-week incubation experiment, high-resolution secondary
ion mass spectrometry imaging of hyphae-associated soil microstructures
revealed a preferential association of 15N with Fe-rich
particles. Synchrotron-based scanning transmission X-ray spectromicroscopy
(SEXM/NEXAFS) of the same samples showed that thin organic coatings
on these soil microstructures are enriched in aliphatic C and
amide N on Fe (hydr)oxides, suggesting a concentration of microbial
lipids and proteins on these surfaces. A possible explanation
for the results of our micro-scale investigation of chemical and
spatial patterns is that amide N from chitinous fungal cell walls
was assimilated by hyphae-associated bacteria, resynthesized into
proteinaceous amide N, and subsequently concentrated onto Fe (hydr)oxide
surfaces. If confirmed in other soil ecosystems, such rapid association
of microbial N with hydroxylated Fe oxide surfaces may have important
implications for mechanistic models of microbial cycling of C
and N.』
1. Introduction
2. Materials and methods
2.1. Sample characteristics
2.1.1. Soil characteristics
2.1.2. Soil incubations
2.1.3. Specimen preparation
2.2. Synchrotron-based scanning transmission X-ray microscopy
(SRXM) in combination with near edge X-ray absorption fine structure
(NEXAFS) spectroscopy
2.2.1. Imaging analysis
2.2.2. Data processing
2.3. High-resolution secondary ion mass spectrometry imaging
2.3.1. Imaging analysis
2.3.2. Data processing
3. Results
3.1. Selection of regions of interest (ROIs)
3.2. NanoSIMS imaging of the δ15N and Fe distribution
3.3. STXM/NEXAFS characterization of δ15N-enriched
features on Fe-rich surfaces
3.3.1. C NEXAFS spectroscopy
3.3.2. N NEXAFS spectroscopy
3.3.3. Fe NEXAFS spectroscopy
3.4. STXM/NEXAFS characterization of spatial and chemical patterns
in hyphae-associated microstructures
3.4.1. C NEXAFS spectroscopy
3.4.2. N NEXAFS spectroscopy
3.5. Characterization of the δ15N and Fe distribution
of distinct morphological and chemical features
4. Discussion
4.1. Microbial C cycling in hyphal-dominated microenvironments
4.2. Microbial amide N cycling in hyphal-dominated microenvironments
4.3. High affinity of Fe (hydr)oxides for microbial C and N
5. Conclusions
Acknowledgments
Appendix A
References