期刊
ANALYTICAL CHEMISTRY
卷 91, 期 3, 页码 2239-2246出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.8b04820
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资金
- National Key Research and Development Program of China [2017YFD0200201]
- Natural Science Foundation of China [21777154, 91851101]
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDB15020302, XDB15020402]
- K. C. Wong Education Foundation
Increasing the bioavailability of immobilized phosphorus (P) in soil by phosphate-solubilizing bacteria (PSB) is an effective strategy for sustainable agronomic use of P and for mitigating the P crisis. Here, D2O isotope labeling combined with single-cell Raman spectroscopy (Raman-D2O) was developed as an efficient activity-based approach to characterizing the presence and activity of PSB in a culture independent way. On the basis of the finding that PSB were significantly more active than non-PSB in the presence of insoluble P, a C-D Raman band from active assimilation of D2O-derived D was established as a biomarker for both inorganic-phosphate-solubilizing bacteria and organic-phosphate-solubilizing bacteria. C-D ratios (intensities of C-D bands as percentages of the intensities of both the C-D and C-H bands) were further established as semiquantitative indicators of P-releasing activities because of the consistency between the C-D ratio and the concentration of solubilized phosphate or acid phosphatase activity as measured by conventional bulk assays. By applying Raman imaging, single-cell Raman-D2O clearly discerned PSB in a mixed-soil bacterial culture and even in complex soil communities. Remarkable heterogeneity of microbial activity, ranging from 2 to 30% (close to that in medium without P and that in medium with sufficient soluble P, respectively), was revealed at the single-cell level and clearly illustrated the subpopulation of soil bacteria active in solubilizing P. This work not only enables probing PSB and their P-releasing activities but also opens a window to explore more diverse microbial resources when obtaining related isotope-labeled substrates is prohibitive.
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