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Article

Microfabrication of a Chamber for High-Resolution, In Situ Imaging of the Whole Root for Plant–Microbe Interactions

1
Environmental Genomics and Systems Biology, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
2
CBRN Defense and Energy Technologies, Sandia National Laboratory, Livermore, CA 94550, USA
3
Joint Genome Institute, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
4
Engineering Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA
*
Authors to whom correspondence should be addressed.
Academic Editor: Eugene S. Vysotski
Int. J. Mol. Sci. 2021, 22(15), 7880; https://doi.org/10.3390/ijms22157880
Received: 30 June 2021 / Revised: 17 July 2021 / Accepted: 18 July 2021 / Published: 23 July 2021
Fabricated ecosystems (EcoFABs) offer an innovative approach to in situ examination of microbial establishment patterns around plant roots using nondestructive, high-resolution microscopy. Previously high-resolution imaging was challenging because the roots were not constrained to a fixed distance from the objective. Here, we describe a new ‘Imaging EcoFAB’ and the use of this device to image the entire root system of growing Brachypodium distachyon at high resolutions (20×, 40×) over a 3-week period. The device is capable of investigating root–microbe interactions of multimember communities. We examined nine strains of Pseudomonas simiae with different fluorescent constructs to B. distachyon and individual cells on root hairs were visible. Succession in the rhizosphere using two different strains of P. simiae was examined, where the second addition was shown to be able to establish in the root tissue. The device was suitable for imaging with different solid media at high magnification, allowing for the imaging of fungal establishment in the rhizosphere. Overall, the Imaging EcoFAB could improve our ability to investigate the spatiotemporal dynamics of the rhizosphere, including studies of fluorescently-tagged, multimember, synthetic communities. View Full-Text
Keywords: imaging; rhizosphere; microfabrication; microscopy; plant–microbe interactions; fluorescently-tagged bacteria; GFP-like proteins imaging; rhizosphere; microfabrication; microscopy; plant–microbe interactions; fluorescently-tagged bacteria; GFP-like proteins
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MDPI and ACS Style

Jabusch, L.K.; Kim, P.W.; Chiniquy, D.; Zhao, Z.; Wang, B.; Bowen, B.; Kang, A.J.; Yoshikuni, Y.; Deutschbauer, A.M.; Singh, A.K.; Northen, T.R. Microfabrication of a Chamber for High-Resolution, In Situ Imaging of the Whole Root for Plant–Microbe Interactions. Int. J. Mol. Sci. 2021, 22, 7880. https://doi.org/10.3390/ijms22157880

AMA Style

Jabusch LK, Kim PW, Chiniquy D, Zhao Z, Wang B, Bowen B, Kang AJ, Yoshikuni Y, Deutschbauer AM, Singh AK, Northen TR. Microfabrication of a Chamber for High-Resolution, In Situ Imaging of the Whole Root for Plant–Microbe Interactions. International Journal of Molecular Sciences. 2021; 22(15):7880. https://doi.org/10.3390/ijms22157880

Chicago/Turabian Style

Jabusch, Lauren K., Peter W. Kim, Dawn Chiniquy, Zhiying Zhao, Bing Wang, Benjamin Bowen, Ashley J. Kang, Yasuo Yoshikuni, Adam M. Deutschbauer, Anup K. Singh, and Trent R. Northen. 2021. "Microfabrication of a Chamber for High-Resolution, In Situ Imaging of the Whole Root for Plant–Microbe Interactions" International Journal of Molecular Sciences 22, no. 15: 7880. https://doi.org/10.3390/ijms22157880

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