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Proceedings Paper

Quantitative 3-dimensional imaging of auxin and cytokinin levels in transgenic soybean and medicago truncatula roots via two-photon induced fluorescence imaging
Author(s): Jon Fisher; Paul Gaillard; Ni Putu Dewi Nurmalasari; Carl Fellbaum; Sen Subramaniam; Steve Smith
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Paper Abstract

Industrial nitrogen fertilizers account for nearly 50% of the fossil fuel costs in modern agriculture and contribute to soil and water pollution. Therefore, significant interest exists in understanding and characterizing the efficiency of nitrogen fixation, and the biochemical signaling pathways which orchestrate the plant-microbial symbiosis through which plants fix nitrogen. Legume plant species exhibit a particularly efficient nitrogen uptake mechanism, using root nodules which house nitrogen-fixing rhizobial bacteria. While nodule development has been widely studied, there remain significant gaps in understanding the regulatory hormones’ role in plant development. In this work, we produce 3-dimensional maps of auxin (AX) and cytokinin (CK) hormone concentrations within model plant root tips and nodules with respect to root architecture and cell type. Soybean and Medicago plants were transfected with a two-color fluorescent vector with AXsensitive green fluorescent protein (GFP) and CK-sensitive TdTomato (TdT). 3D images of soybean root nodules were captured using two-photon induced fluorescence microscopy. The resulting images were computationally analyzed using the localization code first developed by Weeks and later adapted by Kilfoil, and analyzed in the context of the root architecture. Statistical analysis of the resulting 3D hormone level maps reproduce-well the known roles of AX and CK in developing plant roots, and are the first quantitative description of these regulatory hormones tied to specific plant architecture. The analytical methods used, and the spatial distribution of these key regulatory hormones in plant roots, nodule primordia and root nodules, and their statistical interpretation are presented.

Paper Details

Date Published: 23 February 2018
PDF: 6 pages
Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 104982W (23 February 2018); doi: 10.1117/12.2291288
Show Author Affiliations
Jon Fisher, South Dakota School of Mines and Technology (United States)
Paul Gaillard, South Dakota State Univ. (United States)
Ni Putu Dewi Nurmalasari, South Dakota School of Mines and Technology (United States)
Carl Fellbaum, South Dakota State Univ. (United States)
Sen Subramaniam, South Dakota State Univ. (United States)
Steve Smith, South Dakota School of Mines and Technology (United States)

Published in SPIE Proceedings Vol. 10498:
Multiphoton Microscopy in the Biomedical Sciences XVIII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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