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Functional characterisation of metal(loid) processes in planta through the integration of synchrotron techniques and plant molecular biology

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Abstract

Functional characterisation of the genes regulating metal(loid) homeostasis in plants is a major focus for phytoremediation, crop biofortification and food security research. Recent advances in X-ray focussing optics and fluorescence detection have greatly improved the potential to use synchrotron techniques in plant science research. With use of methods such as micro X-ray fluorescence mapping, micro computed tomography and micro X-ray absorption near edge spectroscopy, metal(loids) can be imaged in vivo in hydrated plant tissues at submicron resolution, and laterally resolved metal(loid) speciation can also be determined under physiologically relevant conditions. This article focuses on the benefits of combining molecular biology and synchrotron-based techniques. By using molecular techniques to probe the location of gene expression and protein production in combination with laterally resolved synchrotron techniques, one can effectively and efficiently assign functional information to specific genes. A review of the state of the art in this field is presented, together with examples as to how synchrotron-based methods can be combined with molecular techniques to facilitate functional characterisation of genes in planta. The article concludes with a summary of the technical challenges still remaining for synchrotron-based hard X-ray plant science research, particularly those relating to subcellular level research.

Elemental distribution in Arabidopsis seeds collected by synchrotron micro-XRF

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Acknowledgements

This work was supported by grants from the National Institute of Environmental Health Sciences, Superfund Research Program (grant nos. P42 ES007373-17, NIEHS P20ES01817-02 and EPA RD-83459901-1) to T.P. and M.L.G. and also from the Department of Energy Office of Basic Energy Sciences (grant no. DE-FG02-06ER15809) to M.L.G. A portion of this was work conducted at beamline 2-ID-D at the Advanced Photon Source. Use of the Advanced Photon Source at Argonne National Laboratory was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under contract no. DE-AC02-06CH11357. A portion of this work was performed at beamline X26A, National Synchrotron Light Source (NSLS), Brookhaven National Laboratory. X26A is supported by the Department of Energy (DOE)—Geosciences (DE-FG02-92ER14244 to the University of Chicago—CARS) and DOE Office of Biological and Environmental Research, Environmental Remediation Sciences Division (DE-FC09-96-SR18546 to the University of Kentucky). Use of the NSLS was supported by the DOE under contract no. DEAC02-98CH10886. Part of this work was undertaken on the X-ray fluorescence microprobe (XFM) beamline at the Australian Synchrotron, Victoria, Australia.

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Correspondence to Erica Donner or Tracy Punshon.

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Published in the special paper collection Elemental Imaging and Speciation in Plant Science with guest editors J. Feldmann and E. Krupp.

Erica Donner and Tracy Punshon contributed equally to this article.

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Donner, E., Punshon, T., Guerinot, M.L. et al. Functional characterisation of metal(loid) processes in planta through the integration of synchrotron techniques and plant molecular biology. Anal Bioanal Chem 402, 3287–3298 (2012). https://doi.org/10.1007/s00216-011-5624-9

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