Resolving colocalization of bacteria and metal(loid)s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF)
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Final Accepted Manuscript
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Univ Arizona, Dept Soil Water & Environm SciIssue Date
2016-12
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ELSEVIER SCIENCE BVCitation
Honeker, L. K., Root, R. A., Chorover, J., & Maier, R. M. (2016). Resolving colocalization of bacteria and metal (loid) s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF). Journal of microbiological methods, 131, 23-33.Rights
© 2016 Elsevier B.V. All rights reserved.Collection Information
This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.Abstract
Metal(loid)-contamination of the environment due to anthropogenic activities is a global problem. Understanding the fate of contaminants requires elucidation of biotic and abiotic factors that influence metal(loid) speciation from molecular to field scales. Improved methods are needed to assess micro-scale processes, such as those occurring at biogeochemical interfaces between plant tissues, microbial cells, and metal(loid)s. Here we present an advanced method that combines fluorescence in situ hybridization (FISH) with synchrotron-based multiple-energy micro-focused X-ray fluorescence microprobe imaging (ME pXRF) to examine colocalization of bacteria and metal(loid)s on root surfaces of plants used to phytostabilize metalliferous mine tailings. Bacteria were visualized on a small root section using SytoBC nucleic acid stain and FISH probes targeting the domain Bacteria and a specific group (Alphaproteobacteria, Gammaproteobacteria, or Actinobacteria). The same root region was then analyzed for elemental distribution and metal(loid) speciation of As and Fe using ME pXRF. The FISH and ME pXRF images were aligned using Image.' software to correlate microbiological and geochemical results. Results from quantitative analysis of colocalization show a significantly higher fraction of As colocalized with Fe-oxide plaques on the root surfaces (fraction of overlap 0.49 +/- 0.19) than to bacteria (0.072 +/- 0.052) (p < 0.05). Of the bacteria that colocalized with metal(loid)s, Actinobacteria, known for their metal tolerance, had a higher correlation with both As and Fe than Alphaproteobacteria or Gammaproteobacteria. This method demonstrates how coupling these micro-techniques can expand our understanding of micro-scale interactions between roots, metal(loid)s and microbes, information that should lead to improved mechanistic models of metal(loid) speciation and fate. (C) 2016 Elsevier B.V. All rights reserved.Note
12 month embargo; published online: 29 September 2016ISSN
1872-8359PubMed ID
27693754Version
Final accepted manuscriptSponsors
National Institute of Environmental Health Sciences (NIEHS) Superfund Research Program (SRP) [P42 ES04940, R01 ES1709]; National Science Foundation Graduate Research Fellowhip Program (NSF GRFP) [DGE-1143953]; U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-765F00515]ae974a485f413a2113503eed53cd6c53
10.1016/j.mimet.2016.09.018