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

Authors :
Lauren K. Jabusch
Peter W. Kim
Dawn Chiniquy
Zhiying Zhao
Bing Wang
Benjamin Bowen
Ashley J. Kang
Yasuo Yoshikuni
Adam M. Deutschbauer
Anup K. Singh
Trent R. Northen
Source :
International Journal of Molecular Sciences, Vol 22, Iss 15, p 7880 (2021)
Publication Year :
2021
Publisher :
MDPI AG, 2021.

Abstract

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.

Details

Language :
English
ISSN :
14220067 and 16616596
Volume :
22
Issue :
15
Database :
Directory of Open Access Journals
Journal :
International Journal of Molecular Sciences
Publication Type :
Academic Journal
Accession number :
edsdoj.1c92245b488d469ea1f2c710e02f84c0
Document Type :
article
Full Text :
https://doi.org/10.3390/ijms22157880